2021/04/08 更新

写真a

オオウチ ヒデヨ
大内 淑代
OHUCHI Hideyo
所属
医歯薬学域 教授
職名
教授
外部リンク

研究キーワード

  • 線維芽細胞増殖因子

  • 視物質

  • 網膜

  • 細胞増殖

  • 器官形成

  • オプシン

  • 細胞分化

  • 再生

  • ニワトリ胚

  • 発生

  • 遺伝子

  • マウス

研究分野

  • ライフサイエンス / 解剖学  / 細胞組織学

  • ライフサイエンス / 機能生物化学

  • ライフサイエンス / 発生生物学

 

論文

  • The Opsin 3/Teleost multiple tissue opsin system: mRNA localization in the retina and brain of medaka (Oryzias latipes) 査読

    Sato K, Nwe Nwe K, Ohuchi H

    J Comp Neurol   2021年1月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)  

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  • Fgf10-CRISPR mosaic mutants demonstrate the gene dose-related loss of the accessory lobe and decrease in the number of alveolar type 2 epithelial cells in mouse lung 査読

    Munenori Habuta, Akihiro Yasue, Ken-ichi T. Suzuki, Hirofumi Fujita, Keita Sato, Hitomi Kono, Ayuko Takayama, Tetsuya Bando, Satoru Miyaishi, Seiichi Oyadomari, Eiji Tanaka, Hideyo Ohuchi

    PLOS ONE15 ( 10 ) e0240333 - e0240333   2020年10月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Public Library of Science (PLoS)  

    DOI: 10.1371/journal.pone.0240333

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  • Dkk3/REIC, an N-glycosylated Protein, Is a Physiological Endoplasmic Reticulum Stress Inducer in the Mouse Adrenal Gland. 査読

    Hirofumi Fujita, Tetsuya Bando, Seiichi Oyadomari, Kazuhiko Ochiai, Masami Watanabe, Hiromi Kumon, Hideyo Ohuchi

    Acta medica Okayama74 ( 3 ) 199 - 208   2020年6月

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    担当区分:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Dickkopf 3 (Dkk3) is a secreted protein belonging to the Dkk family and encoded by the orthologous gene of REIC. Dkk3/REIC is expressed by mouse and human adrenal glands, but the understanding of its roles in this organ is still limited. To determine the functions of Dkk3 in the mouse adrenal gland, we first identified that the mouse Dkk3 protein is N-glycosylated in the adrenal gland as well as in the brain. We performed proteome analysis on adrenal glands from Dkk3-null mice, in which exons 5 and 6 of the Dkk3 gene are deleted. Twodimensional polyacrylamide gel electrophoresis of adrenal proteins from wild-type and Dkk3-null mice revealed 5 protein spots whose intensities were altered between the 2 genotypes. Mass spectrometry analysis of these spots identified binding immunoglobulin protein (BiP), an endoplasmic reticulum (ER) chaperone. To determine whether mouse Dkk3 is involved in the unfolded protein response (UPR), we carried out a reporter assay using ER-stress responsive elements. Forced expression of Dkk3 resulted in the induction of distinct levels of reporter expression, showing the UPR initiated by the ER membrane proteins of activating transcription factor 6 (ATF6) and inositol-requring enzyme 1 (IRE1). Thus, it is possible that Dkk3 is a physiological ER stressor in the mouse adrenal gland.

    DOI: 10.18926/AMO/59950

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  • KCNJ13 Gene Deletion Impairs Cell Alignment and Phagocytosis in Retinal Pigment Epithelium Derived from Human-Induced Pluripotent Stem Cells 査読

    Yuki Kanzaki, Hirofumi Fujita, Keita Sato, Mio Hosokawa, Hiroshi Matsumae, Fumio Shiraga, Yuki Morizane, Hideyo Ohuchi

    Investigative Opthalmology & Visual Science61 ( 5 ) 38 - 38   2020年5月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Association for Research in Vision and Ophthalmology (ARVO)  

    DOI: 10.1167/iovs.61.5.38

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  • Dickkopf3 (Dkk3) is required for maintaining the integrity of secretory vesicles in the mouse adrenal medulla. 査読

    Habuta M, Fujita H, Sato K, Bando T, Inoue J, Kondo Y, Miyaishi S, Kumon H, Ohuchi H

    Cell and tissue research379 ( 1 ) 157 - 167   2019年10月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s00441-019-03113-8

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  • Localization of the ultraviolet-sensor Opn5m and its effect on myopia-related gene expression in the late-embryonic chick eye. 査読

    Kato M, Sato K, Habuta M, Fujita H, Bando T, Morizane Y, Shiraga F, Miyaishi S, Ohuchi H

    Biochemistry and biophysics reports19   100665   2019年9月

  • Evolutionary history of teleost intron-containing and intron-less rhodopsin genes. 査読

    Fujiyabu C, Sato K, Utari NML, Ohuchi H, Shichida Y, Yamashita T

    Scientific reports9 ( 1 ) 10653   2019年7月

  • 「光受容体」Opn5が示す多様な分子機能 招待 査読

    佐藤恵太, 山下高廣, 大内淑代, 七田芳則

    生物物理59 ( 3 ) 132 - 136   2019年5月

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  • Glutathione accelerates osteoclast differentiation and inflammatory bone destruction. 査読

    Fujita H, Ochi M, Ono M, Aoyama E, Ogino T, Kondo Y, Ohuchi H

    Free radical research53 ( 2 ) 226 - 236   2019年2月

  • Opn5L1 is a retinal receptor that behaves as a reverse and self-regenerating photoreceptor 査読

    Keita Sato, Takahiro Yamashita, Hideyo Ohuchi, Atsuko Takeuchi, Hitoshi Gotoh, Katsuhiko Ono, Misao Mizuno, Yasuhisa Mizutani, Sayuri Tomonari, Kazumi Sakai, Yasushi Imamoto, Akimori Wada, Yoshinori Shichida

    Nature Communications9 ( 1 ) 1255   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Nature Publishing Group  

    Most opsins are G protein-coupled receptors that utilize retinal both as a ligand and as a chromophore. Opsins' main established mechanism is light-triggered activation through retinal 11-cis-to-all-trans photoisomerization. Here we report a vertebrate non-visual opsin that functions as a Gi-coupled retinal receptor that is deactivated by light and can thermally self-regenerate. This opsin, Opn5L1, binds exclusively to all-trans-retinal. More interestingly, the light-induced deactivation through retinal trans-to-cis isomerization is followed by formation of a covalent adduct between retinal and a nearby cysteine, which breaks the retinal-conjugated double bond system, probably at the C11 position, resulting in thermal re-isomerization to all-trans-retinal. Thus, Opn5L1 acts as a reverse photoreceptor. We conclude that, like vertebrate rhodopsin, Opn5L1 is a unidirectional optical switch optimized from an ancestral bidirectional optical switch, such as invertebrate rhodopsin, to increase the S/N ratio of the signal transduction, although the direction of optimization is opposite to that of vertebrate rhodopsin.

    DOI: 10.1038/s41467-018-03603-3

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  • Bone morphogenetic protein signaling in distal patterning and intercalation during leg regeneration of the cricket, Gryllus bimaculatus. 査読

    Ishimaru Y, Bando T, Ohuchi H, Noji S, Mito T

    Development, growth & differentiation60 ( 6 ) 377 - 386   2018年8月

  • Pinopsin evolved as the ancestral dim-light visual opsin in vertebrates. 査読

    Sato K, Yamashita T, Kojima K, Sakai K, Matsutani Y, Yanagawa M, Yamano Y, Wada A, Iwabe N, Ohuchi H, Shichida Y

    Communications biology1   156   2018年

  • Expression analysis of Dickkopf-related protein 3 (Dkk3) suggests its pleiotropic roles for a secretory glycoprotein in adult mouse 査読

    Junji Inoue, Hirofumi Fujita, Tetsuya Bando, Yoichi Kondo, Hiromi Kumon, Hideyo Ohuchi

    JOURNAL OF MOLECULAR HISTOLOGY48 ( 1 ) 29 - 39   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    Dickkopf-related protein 3 (Dkk3) is the third member of the Dkk gene family and identical to the gene, whose expression was reduced in immortalized cells. Therefore, its another name is reduced expression in immortalized cells. Since the intratumoral introduction of Dkk3 inhibits tumor growth in mouse models of cancers, Dkk3 is likely a tumor suppressor gene. However, the functions of Dkk3 in vivo remain unclear. As the first step to decipher the physiological roles of this gene, we examined the expression pattern of Dkk3 in various tissues from adult mice. In situ hybridization showed that Dkk3 mRNA was detected in the brain, retina, heart, gastrointestinal tract, adrenal glands, thymus, prostate glands, seminal vesicles, testes, and ovaries in a regionally specific manner. Furthermore, we raised anti-mouse Dkk3 antibody and performed immunohistochemistry. Cytoplasmic localization of Dkk3 protein was observed in the cells of the adrenal medulla, while Dkk3 immunoreactivity was observed in the lumen of the stomach and intestine, implying that the Dkk3 protein may be secreted into the lumen of the gastrointestinal tract. These results suggest that Dkk3 has pleiotropic roles for a secretory glycoprotein that acts primarily in the gastrointestinal tract, thymus, endocrine and reproductive organs of the mouse.

    DOI: 10.1007/s10735-016-9703-2

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  • Eye development and photoreception of a hemimetabolous insect, gryllus bimaculatus 招待 査読

    Hideyo Ohuchi, Tetsuya Bando, Taro Mito, Sumihare Noji

    The Cricket as a Model Organism: Development, Regeneration, and Behavior   49 - 62   2017年1月

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:論文集(書籍)内論文   出版者・発行元:Springer Japan  

    The hemimetabolous insect, Gryllus bimaculatus, has two compound eyes that begin to form in the embryo and increase in size five- to sixfolds during nymphal development. Retinal stemlike cells reside in the anteroventral proliferation zone (AVPZ) of the nymphal compound eye and proliferate to increase retinal progenitors, which then differentiate to form new ommatidia in the anterior region of the eye. Here, we introduce the morphology and development of the cricket eye first, and then we focus on the roles of retinal determination genes (RDGs) such as eyes absent (eya) and sine oculis (so) in Gryllus eye formation and growth. Since the principal function of the eye is photoreception, we finally summarize opsin photopigments in this species, broadening the roles of photoreception.

    DOI: 10.1007/978-4-431-56478-2_4

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  • Relationship between somatic mosaicism of Pax6 mutation and variable developmental eye abnormalities-an analysis of CRISPR genome-edited mouse embryos 査読

    Akihiro Yasue, Hitomi Kono, Munenori Habuta, Tetsuya Bando, Keita Sato, Junji Inoue, Seiichi Oyadomari, Sumihare Noji, Eiji Tanaka, Hideyo Ohuchi

    Scientific Reports7 ( 1 ) 53   2017年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Nature Publishing Group  

    The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-Associated protein (Cas) system is a rapid gene-Targeting technology that does not require embryonic stem cells. To demonstrate dosage effects of the Pax6 gene on eye formation, we generated Pax6-deficient mice with the CRISPR/Cas system. Eyes of founder embryos at embryonic day (E) 16.5 were examined and categorized according to macroscopic phenotype as class 1 (small eye with distinct pigmentation), class 2 (pigmentation without eye globes), or class 3 (no pigmentation and no eyes). Histologically, class 1 eyes were abnormally small in size with lens still attached to the cornea at E16.5. Class 2 eyes had no lens and distorted convoluted retinas. Class 3 eyes had only rudimentary optic vesicle-like tissues or histological anophthalmia. Genotyping of neck tissue cells from the founder embryos revealed somatic mosaicism and allelic complexity for Pax6. Relationships between eye phenotype and genotype were developed. The present results demonstrated that development of the lens from the surface ectoderm requires a higher gene dose of Pax6 than development of the retina from the optic vesicle. We further anticipate that mice with somatic mosaicism in a targeted gene generated by CRISPR/Cas-mediated genome editing will give some insights for understanding the complexity in human congenital diseases that occur in mosaic form.

    DOI: 10.1038/s41598-017-00088-w

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  • Two Opsin 3-Related Proteins in the Chicken Retina and Brain: A TMT-Type Opsin 3 Is a Blue-Light Sensor in Retinal Horizontal Cells, Hypothalamus, and Cerebellum 査読

    Mutsuko Kato, Takashi Sugiyama, Kazumi Sakai, Takahiro Yamashita, Hirofumi Fujita, Keita Sato, Sayuri Tomonari, Yoshinori Shichida, Hideyo Ohuchi

    PLOS ONE11 ( 11 ) e0163925   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Opsin family genes encode G protein-coupled seven-transmembrane proteins that bind a retinaldehyde chromophore in photoreception. Here, we sought potential as yet undescribed avian retinal photoreceptors, focusing on Opsin 3 homologs in the chicken. We found two Opsin 3-related genes in the chicken genome: one corresponding to encephalopsin/panopsin (Opn3) in mammals, and the other belonging to the teleost multiple tissue opsin (TMT) 2 group. Bioluminescence imaging and G protein activation assays demonstrated that the chicken TMT opsin (cTMT) functions as a blue light sensor when forced-expressed in mammalian cultured cells. We did not detect evidence of light sensitivity for the chicken Opn3 (cOpn3). In situ hybridization demonstrated expression of cTMT in subsets of differentiating cells in the inner retina and, as development progressed, predominant localization to retinal horizontal cells (HCs). Immunohistochemistry (IHC) revealed cTMT in HCs as well as in small numbers of cells in the ganglion and inner nuclear layers of the post-hatch chicken retina. In contrast, cOpn3-IR cells were found in distinct subsets of cells in the inner nuclear layer. cTMT-IR cells were also found in subsets of cells in the hypothalamus. Finally, we found differential distribution of cOpn3 and cTMT proteins in specific cells of the cerebellum. The present results suggest that a novel TMT-type opsin 3 may function as a photoreceptor in the chicken retina and brain.

    DOI: 10.1371/journal.pone.0163925

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  • TGF-β signaling in insects regulates metamorphosis via juvenile hormone biosynthesis. 査読

    Ishimaru Y, Tomonari S, Matsuoka Y, Watanabe T, Miyawaki K, Bando T, Tomioka K, Ohuchi H, Noji S, Mito T

    Proceedings of the National Academy of Sciences of the United States of America113 ( 20 ) 5634 - 5639   2016年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1073/pnas.1600612113

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  • Two UV-Sensitive Photoreceptor Proteins, Opn5m and Opn5m2 in Ray-Finned Fish with Distinct Molecular Properties and Broad Distribution in the Retina and Brain 査読

    Keita Sato, Takahiro Yamashita, Yoshihiro Haruki, Hideyo Ohuchi, Masato Kinoshita, Yoshinori Shichida

    PLOS ONE11 ( 5 ) e0155339   2016年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Opn5 is a group within the opsin family of proteins that is responsible for visual and non-visual photoreception in animals. It consists of several subgroups, including Opn5m, the only subgroup containing members found in most vertebrates, including mammals. In addition, recent genomic information has revealed that some ray-finned fishes carry paralogous genes of Opn5m while other fishes have no such genes. Here, we report the molecular properties of the opsin now called Opn5m2 and its distributions in both the retina and brain. Like Opn5m, Opn5m2 exhibits UV light-sensitivity when binding to 11-cis-retinal and forms a stable active state that couples with Gi subtype of G protein. However, Opn5m2 does not bind all-trans-retinal and exhibits exclusive binding to 11-cis-retinal, whereas many bistable opsins, including fish Opn5m, can bind directly to all-trans-retinal as well as 11-cis-retinal. Because medaka fish has lost the Opn5m2 gene from its genome, we compared the tissue distribution patterns of Opn5m in medaka fish, zebrafish, and spotted gar, in addition to the distribution patterns of Opn5m2 in zebrafish and spotted gar. Opn5m expression levels showed a gradient along the dorsal-ventral axis of the retina, and preferential expression was observed in the ventral retina in the three fishes. The levels of Opn5m2 showed a similar gradient with preferential expression observed in the dorsal retina. Opn5m expression was relatively abundant in the inner region of the inner nuclear layer, while Opn5m2 was expressed in the outer edge of the inner nuclear layer. Additionally, we could detect Opn5m expression in several brain regions, including the hypothalamus, of these fish species. Opn5m2 expression could not be detected in zebrafish brain, but was clearly observed in limited brain regions of spotted gar. These results suggest that ray-finned fishes can generally utilize UV light information for non-image-forming photoreception in a wide range of cells in the retina and brain.

    DOI: 10.1371/journal.pone.0155339

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  • Autopsy Case of Bilateral Optic Nerve Aplasia with Microphthalmia: Neural Retina Formation Is Required for the Coordinated Development of Ocular Tissues 査読

    Hideyo Ohuchi, Kaori Taniguchi, Satoru Miyaishi, Hitomi Kono, Hirofumi Fujita, Tetsuya Bando, Chiharu Fuchizawa, Yuko Ohtani, Osamu Ohtani

    ACTA MEDICA OKAYAMA70 ( 2 ) 131 - 137   2016年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OKAYAMA UNIV MED SCHOOL  

    Human congenital anomalies provide information that contributes to the understanding of developmental mechanisms. Here we report bilateral optic nerve aplasia (ONA) with microphthalmia in the autopsy of the cadaver of a 70-year-old Japanese female. The gross anatomical inspection of the brain showed a cotton thread-like cord in the presumed location of the optic nerve tract or chiasm. Histologically, no neural retina, optic nerve bundle or retinal central vessels were formed in the eye globe, and the retinal pigment cells formed rosettes. The cornea, iris, and lens were also histologically abnormal. Immunohistochemically, no retinal cells expressed beta III tubulin, and Pax6-immunoreactive cells were present in the ciliary non-pigmented epithelial cells. This case of ONA could be attributed to the agenesis of retinal projection neurons as a sequel to the disruption of neural retina development. The neural retina formation would coordinate the proper development of ocular tissues.

    DOI: 10.18926/AMO/54192

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  • Phytoestrogen Suppresses Efflux of the Diagnostic Marker Protoporphyrin IX in Lung Carcinoma 査読

    Hirofumi Fujita, Keisuke Nagakawa, Hirotsugu Kobuchi, Tetsuya Ogino, Yoichi Kondo, Keiji Inoue, Taro Shuin, Toshihiko Utsumi, Kozo Utsumi, Junzo Sasaki, Hideyo Ohuchi

    CANCER RESEARCH76 ( 7 ) 1837 - 1846   2016年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ASSOC CANCER RESEARCH  

    One promising method to visualize cancer cells is based on the detection of the fluorescent photosensitizer protoporphyrin IX (PpIX) synthesized from 5-aminolevulinic acid (ALA), but this method cannot be used in cancers that exhibit poor PpIX accumulation. PpIX appears to be pumped out of cancer cells by the ABC transporter G2 (ABCG2), which is associated with multidrug resistance. Genistein is a phytoestrogen that appears to competitively inhibit ABCG2 activity. Therefore, we investigated whether genistein can promote PpIX accumulation in human lung carcinoma cells. Here we report that treatment of A549 lung carcinoma cells with genistein or a specific ABCG2 inhibitor promoted ALA-mediated accumulation of PpIX by approximately 2-fold. ABCG2 depletion and overexpression studies further revealed that genistein promoted PpIX accumulation via functional repression of ABCG2. After an extended period of genistein treatment, a significant increase in PpIX accumulation was observed in A549 cells (3.7-fold) and in other cell lines. Systemic preconditioning with genistein in a mouse xenograft model of lung carcinoma resulted in a 1.8-fold increase in accumulated PpIX. Long-term genistein treatment stimulated the expression of genes encoding enzymes involved in PpIX synthesis, such as porphobilinogen deaminase, uroporphyrinogen decarboxylase, and protoporphyrinogen oxidase. Accordingly, the rate of PpIX synthesis was also accelerated by genistein pretreatment. Thus, our results suggest that genistein treatment effectively enhances ALA-induced PpIX accumulation by preventing the ABCG2-mediated efflux of PpIX from lung cancer cells and may represent a promising strategy to improve ALA-based diagnostic approaches in a broader set of malignancies.

    DOI: 10.1158/0008-5472.CAN-15-1484

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  • 昆虫に学ぶ器官再生 招待

    板東哲哉, 奥村美紗, 坂東優希, 大内淑代

      ( 3 ) 2 - 6   2016年

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    掲載種別:研究論文(学術雑誌)  

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  • Enhancer of zeste plays an important role in photoperiodic modulation of locomotor rhythm in the cricket, Gryllus bimaculatus. 査読 国際誌

    Yoshimasa Hamada, Atsushi Tokuoka, Tetsuya Bando, Hideyo Ohuchi, Kenji Tomioka

    Zoological letters2   5 - 5   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    INTRODUCTION: Insects show daily behavioral rhythms controlled by an endogenous oscillator, the circadian clock. The rhythm synchronizes to daily light-dark cycles (LD) and changes waveform in association with seasonal change in photoperiod. RESULTS: To explore the molecular basis of the photoperiod-dependent changes in circadian locomotor rhythm, we investigated the role of a chromatin modifier, Enhancer of zeste (Gb'E(z)), in the cricket, Gryllus bimaculatus. Under a 12 h:12 h LD (LD 12:12), Gb'E(z) was constitutively expressed in the optic lobe, the site of the biological clock; active phase (α) and rest phase (ρ) were approximately 12 h in duration, and α/ρ ratio was approximately 1.0. When transferred to LD 20:4, the α/ρ ratio decreased significantly, and the Gb'E(z) expression level was significantly reduced at 6 h and 10 h after light-on, as was reflected in the reduced level of trimethylation of histone H3 lysine 27. This change was associated with change in clock gene expression profiles. The photoperiod-dependent changes in α/ρ ratio and clock gene expression profiles were prevented by knocking down Gb'E(z) by RNAi. CONCLUSIONS: These results suggest that histone modification by Gb'E(z) is involved in photoperiodic modulation of the G. bimaculatus circadian rhythm.

    DOI: 10.1186/s40851-016-0042-7

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  • Leg regeneration is epigenetically regulated by histone H3K27 methylation in the cricket Gryllus bimaculatus 査読

    Yoshimasa Hamada, Tetsuya Bando, Taro Nakamura, Yoshiyasu Ishimaru, Taro Mito, Sumihare Noji, Kenji Tomioka, Hideyo Ohuchi

    DEVELOPMENT142 ( 17 ) 2916 - +   2015年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Hemimetabolous insects such as the cricket Gryllus bimaculatus regenerate lost tissue parts using blastemal cells, a population of dedifferentiated proliferating cells. The expression of several factors that control epigenetic modification is upregulated in the blastema compared with differentiated tissue, suggesting that epigenetic changes in gene expression might control the differentiation status of blastema cells during regeneration. To clarify the molecular basis of epigenetic regulation during regeneration, we focused on the function of the Gryllus Enhancer of zeste [Gb'E(z)] and Ubiquitously transcribed tetratricopeptide repeat gene on the X chromosome (Gb'Utx) homologues, which regulate methylation and demethylation of histone H3 lysine 27 (H3K27), respectively. Methylated histone H3K27 in the regenerating leg was diminished by Gb'E(z)(RNAi) and was increased by Gb'Utx(RNAi). Regenerated Gb'E(z)(RNAi) cricket legs exhibited extra leg segment formation between the tibia and tarsus, and regenerated Gb'Utx(RNAi) cricket legs showed leg joint formation defects in the tarsus. In the Gb'E(z)(RNAi) regenerating leg, the Gb'dac expression domain expanded in the tarsus. By contrast, in the Gb'Utx(RNAi) regenerating leg, Gb'Egfr expression in the middle of the tarsus was diminished. These results suggest that regulation of the histone H3K27 methylation state is involved in the repatterning process during leg regeneration among cricket species via the epigenetic regulation of leg patterning gene expression.

    DOI: 10.1242/dev.122598

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  • Involvement of dachshund and Distal-less in distal pattern formation of the cricket leg during regeneration 査読

    Yoshiyasu Ishimaru, Taro Nakamura, Tetsuya Bando, Yuji Matsuoka, Hideyo Ohuchi, Sumihare Noji, Taro Mito

    SCIENTIFIC REPORTS5   8387   2015年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Cricket nymphs have the remarkable ability to regenerate a functional leg following amputation, indicating that the regenerating blastemal cells contain information for leg morphology. However, the molecular mechanisms that underlie regeneration of leg patterns remain poorly understood. Here, we analyzed phenotypes of the tibia and tarsus (three tarsomeres) obtained by knockdown with regeneration-dependent RNA interference (rdRNAi) against Gryllus dachshund (Gb'dac) and Distal-less (Gb'Dll). We found that depletion of Gb'Dll mRNA results in loss of the tarsal segments, while rdRNAi against Gb'dac shortens the tibia at the two most distal tarsomeres. These results indicate that Gb'Dll expression is indispensable for formation of the tarsus, while Gb'dac expression is necessary for elongation of the tibia and formation of the most proximal tarsomere. These findings demonstrate that mutual transcriptional regulation between the two is indispensable for formation of the tarsomeres, whereas Gb'dac is involved in determination of tibial size through interaction with Gb'ds/Gb'ft.

    DOI: 10.1038/srep08387

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  • An extended steepness model for leg-size determination based on Dachsous/Fat trans- dimer system 査読

    Hiroshi Yoshida, Tetsuya Bando, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    SCIENTIFIC REPORTS4   4335   2014年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    What determines organ size has been a long-standing biological question. Lawrence et al. (2008) proposed the steepness hypothesis suggesting that the protocadherin Dachsous/Fat (Ds/Ft) system may provide some measure of dimension to the cells in relation to the gradient. In this paper we extended the model as a means of interpreting experimental results in cricket leg regeneration. We assumed that (1) Ds/Ft trans-heterodimers or trans-homodimers are redistributed during cell division, and (2) growth would cease when a differential of the dimer across each cell decreases to a certain threshold. We applied our model to simulate the results obtained by leg regeneration experiments in a cricket model. The results were qualitatively consistent with the experimental data obtained for cricket legs by RNA interference methodology. Using our extended steepness model, we provided a molecular-based explanation for leg size determination even in intercalary regeneration and for organ size determination.

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  • Evolution of Mammalian Opn5 as a Specialized UV-absorbing Pigment by a Single Amino Acid Mutation 査読

    Takahiro Yamashita, Katsuhiko Ono, Hideyo Ohuchi, Akane Yumoto, Hitoshi Gotoh, Sayuri Tomonari, Kazumi Sakai, Hirofumi Fujita, Yasushi Imamoto, Sumihare Noji, Katsuki Nakamura, Yoshinori Shichida

    JOURNAL OF BIOLOGICAL CHEMISTRY289 ( 7 ) 3991 - 4000   2014年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Opn5 is one of the recently identified opsin groups that is responsible for nonvisual photoreception in animals. We previously showed that a chicken homolog of mammalian Opn5 (Opn5m) is a G(i)-coupled UV sensor having molecular properties typical of bistable pigments. Here we demonstrated that mammalian Opn5m evolved to be a more specialized photosensor by losing one of the characteristics of bistable pigments, direct binding of all-trans-retinal. We first confirmed that Opn5m proteins in zebrafish, Xenopus tropicalis, mouse, and human are also UV-sensitive pigments. Then we found that only mammalian Opn5m proteins lack the ability to directly bind all-trans-retinal. Mutational analysis showed that these characteristics were acquired by a single amino acid replacement at position 168. By comparing the expression patterns of Opn5m between mammals and chicken, we found that, like chicken Opn5m, mammalian Opn5m was localized in the ganglion cell layer and inner nuclear layer of the retina. However, the mouse and primate (common marmoset) opsins were distributed not in the posterior hypothalamus (including the region along the third ventricle) where chicken Opn5m is localized, but in the preoptic hypothalamus. Interestingly, RPE65, an essential enzyme for forming 11-cis-retinal in the visual cycle is expressed near the preoptic hypothalamus of the mouse and common marmoset brain but not near the region of the chicken brain where chicken Opn5m is expressed. Therefore, mammalian Opn5m may work exclusively as a short wavelength sensor in the brain as well as in the retina with the assistance of an 11-cis-retinal-supplying system.

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  • The expression of LIM-homeobox genes, Lhx1 and Lhx5, in the forebrain is essential for neural retina differentiation 査読

    Junji Inoue, Yuuki Ueda, Tetsuya Bando, Taro Mito, Sumihare Noji, Hideyo Ohuchi

    DEVELOPMENT GROWTH & DIFFERENTIATION55 ( 7 ) 668 - 675   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Elucidating the mechanisms underlying eye development is essential for advancing the medical treatment of eye-related disorders. The primordium of the eye is an optic vesicle (OV), which has a dual potential for generation of the developing neural retina and retinal pigment epithelium. However, the factors that regulate the differentiation of the retinal primordium remain unclear. We have previously shown that overexpression of Lhx1 and Lhx5, members of the LIM-homeobox genes, induced the formation of a second neural retina from the presumptive pigmented retina of the OV. However, the precise timing of Lhx1 expression required for neural retina differentiation has not been clarified. Moreover, RNA interference of Lhx5 has not been previously reported. Here, using a modified electroporation method, we show that, Lhx1 expression in the forebrain around stage 8 is required for neural retina formation. In addition, we have succeeded in the knockdown of Lhx5 expression, resulting in conversion of the neural retina region to a pigment vesicle-like tissue, which indicates that Lhx5 is also required for neural retina differentiation, which correlates temporally with the activity of Lhx1. These results suggest that Lhx1 and Lhx5 in the forebrain regulate neural retina differentiation by suppressing the development of the retinal pigment epithelium, before the formation of the OV.

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  • Improvement of the Efficacy of 5-aminolevulinic Acid-mediated Photodynamic Treatment in Human Oral Squamous Cell Carcinoma HSC-4 査読

    Masanao Yamamoto, Hirofumi Fujita, Naoki Katase, Keiji Inoue, Hitoshi Nagatsuka, Kozo Utsumi, Junzo Sasaki, Hideyo Ohuchi

    ACTA MEDICA OKAYAMA67 ( 3 ) 153 - 164   2013年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OKAYAMA UNIV MED SCHOOL  

    Ever since protoporphyrin IX (PpIX) was discovered to accumulate preferentially in cancer cells after 5-aminolevulinic acid (ALA) treatment, photodynamic treatment or therapy (PDT) has been developed as an exciting new treatment option for cancer patients. However, the level of PpIX accumulation in oral cancer is fairly low and insufficient for PDT. Ferrochelatase (FECH) and ATP-binding cassette transporter G2 (ABCG2) are known to regulate PpIX accumulation. In addition, serum enhances PpIX export by ABCG2. We investigated here whether and how inhibitors of FECH and ABCG2 and their combination could improve PpIX accumulation and PDT efficacy in an oral cancer cell line in serum-containing medium. ABCG2 inhibitor and the combination of ABCG2 and FECH inhibitors increased PpIX in the presence of fetal bovine serum (FBS) in an oral cancer cell line. Analysis of ABCG2 gene silencing also revealed the involvement of ABCG2 in the regulation of PpIX accumulation. Inhibitors of FECH and ABCG2, and their combination increased the efficiency of ALA-PDT even in the presence of FBS. ALA-PDT-induced cell death was accompanied by apoptotic events and lipid peroxidation. These results suggest that accumulation of PpIX is determined by the activities of ABCG2 and FECH and that treatment with a combination of their inhibitors improves the efficacy of PDT for oral cancer, especially in the presence of serum.

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  • Analysis of RNA-Seq data reveals involvement of JAK/STAT signalling during leg regeneration in the cricket Gryllus bimaculatus 査読

    Tetsuya Bando, Yoshiyasu Ishimaru, Takuro Kida, Yoshimasa Hamada, Yuji Matsuoka, Taro Nakamura, Hideyo Ohuchi, Sumihare Noji, Taro Mito

    DEVELOPMENT140 ( 5 ) 959 - 964   2013年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    In the cricket Gryllus bimaculatus, missing distal parts of the amputated leg are regenerated from the blastema, a population of dedifferentiated proliferating cells that forms at the distal tip of the leg stump. To identify molecules involved in blastema formation, comparative transcriptome analysis was performed between regenerating and normal unamputated legs. Components of JAK/STAT signalling were upregulated more than twofold in regenerating legs. To verify their involvement, Gryllus homologues of the interleukin receptor Domeless (Gb'dome), the Janus kinase Hopscotch (Gb'hop) and the transcription factor STAT (Gb'Stat) were cloned, and RNAi was performed against these genes. Gb'dome(RNAi), Gb'hop(RNAi) and Gb'Stat(RNAi) crickets showed defects in leg regeneration. Blastema expression of Gb'cyclinE was decreased in the Gb'Stat(RNAi) cricket compared with that in the control. Hyperproliferation of blastema cells caused by Gb'fat(RNAi) or Gb'warts(RNAi) was suppressed by RNAi against Gb'Stat. The results suggest that JAK/STAT signalling regulates blastema cell proliferation during leg regeneration.

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  • Lhx1 in the proximal region of the optic vesicle permits neural retina development in the chicken 査読

    Takumi Kawaue, Mayumi Okamoto, Akane Matsuyo, Junji Inoue, Yuhki Ueda, Sayuri Tomonari, Sumihare Noji, Hideyo Ohuchi

    BIOLOGY OPEN1 ( 11 ) 1083 - 1093   2012年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    How the eye forms has been one of the fundamental issues in developmental biology. The retinal anlage first appears as the optic vesicle (OV) evaginating from the forebrain. Subsequently, its distal portion invaginates to form the two-walled optic cup, which develops into the outer pigmented and inner neurosensory layers of the retina. Recent work has shown that this optic-cup morphogenesis proceeds as a self-organizing activity without any extrinsic molecules. However, intrinsic factors that regulate this process have not been elucidated. Here we show that a LIM-homeobox gene, Lhx1, normally expressed in the proximal region of the nascent OV, induces a second neurosensory retina formation from the outer pigmented retina when overexpressed in the chicken OV. Lhx2, another LIM-homeobox gene supposed to be involved in early OV formation, could not substitute this function of Lhx1, while Lhx5, closely related to Lhx1, could replace it. Conversely, knockdown of Lhx1 expression by RNA interference resulted in the formation of a small or pigmented vesicle. These results suggest that the proximal region demarcated by Lhx1 expression permits OV development, eventually dividing the two retinal domains. (C) 2012. Published by The Company of Biologists Ltd.

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  • Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases 査読

    Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Hadley W. Horch, Naoya Hamaguchi, Taro Nakamura, Tetsuya Bando, Hideyo Ohuchi, Takashi Yamamoto, Sumihare Noji, Taro Mito

    NATURE COMMUNICATIONS3   1017   2012年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically relatively basal and comprise many pests. However, the absence of a sophisticated genetic model system, or targeted gene-manipulation system, has limited research on hemimetabolous species. Here we use zinc-finger nuclease and transcription activator-like effector nuclease technologies to produce genetic knockouts in the hemimetabolous insect Gryllus bimaculatus. Following the microinjection of mRNAs encoding zinc-finger nucleases or transcription activator-like effector nucleases into cricket embryos, targeting of a transgene or endogenous gene results in sequence-specific mutations. Up to 48% of founder animals transmit disrupted gene alleles after zinc-finger nucleases microinjection compared with 17% after microinjection of transcription activator-like effector nucleases. Heterozygous offspring is selected using mutation detection assays that use a Surveyor (Cel-I) nuclease, and subsequent sibling crosses create homozygous knockout crickets. This approach is independent from a mutant phenotype or the genetic tractability of the organism of interest and can potentially be applied to manage insect pests using a non-transgenic strategy.

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  • A Non-Mammalian Type Opsin 5 Functions Dually in the Photoreceptive and Non-Photoreceptive Organs of Birds 査読

    Hideyo Ohuchi, Takahiro Yamashita, Sayuri Tomonari, Sari Fujita-Yanagibayashi, Kazumi Sakai, Sumihare Noji, Yoshinori Shichida

    PLOS ONE7 ( 2 ) e31534   2012年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    A mammalian type opsin 5 (neuropsin) is a recently identified ultraviolet (UV)-sensitive pigment of the retina and other photosensitive organs in birds. Two other opsin 5-related molecules have been found in the genomes of non-mammalian vertebrates. However, their functions have not been examined as yet. Here, we identify the molecular properties of a second avian opsin 5, cOpn5L2 (chicken opsin 5-like 2), and its localization in the post-hatch chicken. Spectrophotometric analysis and radionucleotide-binding assay have revealed that cOpn5L2 is a UV-sensitive bistable pigment that couples with the Gi subtype of guanine nucleotide-binding protein (G protein). As a bistable pigment, it also shows the direct binding ability to agonist all-trans-retinal to activate G protein. The absorption maxima of UV-light-absorbing and visible light-absorbing forms were 350 and 521 nm, respectively. Expression analysis showed relatively high expression of cOpn5L2 mRNA in the adrenal gland, which is not photoreceptive but an endocrine organ, while lower expression was found in the brain and retina. At the protein level, cOpn5L2 immunoreactive cells were present in the chromaffin cells of the adrenal gland. In the brain, cOpn5L2 immunoreactive cells were found in the paraventricular and supraoptic nuclei of the anterior hypothalamus, known for photoreceptive deep brain areas. In the retina, cOpn5L2 protein was localized to subsets of cells in the ganglion cell layer and the inner nuclear layer. These results suggest that the non-mammalian type opsin 5 (Opn5L2) functions as a second UV sensor in the photoreceptive organs, while it might function as chemosensor using its direct binding ability to agonist all-trans-retinal in non-photoreceptive organs such as the adrenal gland of birds.

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  • Functional analysis of the role of eyes absent and sine oculis in the developing eye of the cricket Gryllus bimaculatus 査読

    Akira Takagi, Kazuki Kurita, Taiki Terasawa, Taro Nakamura, Tetsuya Bando, Yoshiyuki Moriyama, Taro Mito, Sumihare Noji, Hideyo Ohuchi

    DEVELOPMENT GROWTH & DIFFERENTIATION54 ( 2 ) 227 - 240   2012年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    In the cricket Gryllus bimaculatus, a hemimetabolous insect, the compound eyes begin to form in the embryo and increase 56 fold in size during the postembryonic development of the nymphal stage. Retinal stem cells in the anteroventral proliferation zone (AVPZ) of the nymphal eye proliferate to increase retinal progenitors, which then differentiate to form new ommatidia in the anterior region of the eye. However, mechanisms underlying this type of eye formation have not been well elucidated yet. Here, we found that the homologues of the retinal determination transcription factor genes of eyes absent (eya) and sine oculis (so) are expressed during the cricket embryonic eye formation. eya is also expressed intensely in the AVPZ of the nymphal eye. To explore their functions, we performed knockdown by RNA interference (RNAi). Knockdown of Gbeya resulted in loss of the embryonic eye. In the nymphal eye, RNAi against Gbeya or Gbso impaired retinal morphology by apparently transforming cornea structures into head cuticle. These results imply that Gbeya and Gbso are essential for the differentiation of the retinal progenitor cells and maintaining retinal structures during eye development.

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  • Establishment of a novel system to elucidate the mechanisms underlying light-induced ripening of strawberry fruit with an Agrobacterium-mediated RNAi technique 査読

    Katsuyuki Miyawaki, Sachi Fukuoka, Yasuko Kadomura, Hirokazu Hamaoka, Taro Mito, Hideyo Ohuchi, Wilfried Schwab, Sumihare Noji

    PLANT BIOTECHNOLOGY29 ( 3 ) 271 - 277   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE SOC PLANT CELL & MOLECULAR BIOLOGY  

    Traditional methods used to study strawberry ripening-related gene function are time-consuming, and require at least 15 months from initiating the transformation experiment until the first ripe fruits are available for analysis. To accelerate data acquisition during gene function studies, we explored a transient assay method that employs an Agrobacterium-mediated RNAi (AmRNAi) technique in post-harvest strawberry fruit, Fragaria x ananassa (Fa) cv. Sachinoka, a Japanese cultivar. Our results showed that artificial white light induced strong expression of Fa'chalcone synthase (Fa'CHS), Fa'chalcone isomerase (Fa'CHI), and Fa'flavonoid 3'-hydroxylase orthologues (Fa'F3'H) in post-harvest fruit. Fa'CHS and Fa'F3'H function was subsequently examined by performing AmRNAi with post-harvest fruit. Although reduction of light-induced Fa'F3'H expression by AmRNAi resulted in no significant change in anthocyanin content, reduction of Fa'CHS significantly decreased anthocyanin levels, and up-regulated Fa'F3'H levels. Our results are consistent with previous data indicating that while CHS is required for anthocyanin accumulation during late stage strawberry fruit maturation, Fa'F3'H is not required. The novel system described here enabled gene function data to be available within 10 days of initiating the incubation period following in filtration. therefore, we conclude our system is a valuable tool to elucidate the molecular mechanisms underlying light-induced ripening of strawberry fruit.

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  • Expression of myelin genes in the developing chick retina 査読

    Hitosh Gotoh, Takayuki Ueda, Aoi Uno, Hideyo Ohuchi, Kazuhiro Ikenaka, Katsuhiko Ono

    GENE EXPRESSION PATTERNS11 ( 8 ) 471 - 475   2011年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    In submammalian animals including chicks, the retina contains oligodendrocytes (OLs), and axons in the optic fiber layer are wrapped with compact myelin within the retina; however, the expression of myelin genes in the chick retina has not been demonstrated yet. In the present study, we examined the expression of three myelin genes (proteolipid protein, PLP; myelin basic protein. MBP; cyclic nucleotide phosphodiesterase, CNP) and PLP in the developing chick retina, in comparison to the localization of Mueller cells. In situ hybridization demonstrated that all three myelin genes began to be expressed at E14 in the chick embryo retina. They are mostly restricted to the ganglion cell layer and the optic fiber layer, with a few exceptions in the inner nuclear layer where Mueller cells reside; however. PLP mRNA+ cells do not express glutamine synthetase, or vice versa. The present results elucidate that myelin genes are expressed only by OLs that are mostly localized in the innermost layer of the developing chick retina. (C) 2011 Elsevier B.V. All rights reserved.

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  • Vertebrate Ancient-Long Opsin Has Molecular Properties Intermediate between Those of Vertebrate and Invertebrate Visual Pigments 査読

    Keita Sato, Takahiro Yamashita, Hideyo Ohuchi, Yoshinori Shichida

    BIOCHEMISTRY50 ( 48 ) 10484 - 10490   2011年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    VA/VAL opsin is one of the four kinds of nonvisual opsins that are closely related to vertebrate visual pigments in the phylogenetic tree of opsins. Previous studies indicated that among these opsins, parapinopsin and pinopsin exhibit molecular properties similar to those of invertebrate bistable visual pigments and vertebrate visual pigments, respectively. Here we show that VA/VAL opsin exhibits molecular properties intermediate between those of parapinopsin and pinopsin. VAL opsin from Xenopus tropicalis was expressed in cultured cells, and the pigment with an absorption maximum at 501 nm was reconstituted by incubation with 11-cis-retinal. Light irradiation of this pigment caused cis-to-trans isomerization of the chromophore to form a state having an absorption maximum in the visible region. This state has the ability to activate Gi and Gt types of G proteins. Therefore, the active state of VAL opsin is a visible light-absorbing intermediate, which probably has a protonated retinylidene Schiff base as its chromophore, like the active state of parapinopsin. However, this state was apparently photoinsensitive and did not show reverse reaction to the original pigment, unlike the active state of parapinopsin, and instead similar to that of pinopsin. Furthermore, the Gi activation efficiency of VAL opsin was between those of pinopsin and parapinopsin. Thus, the molecular properties of VA/VAL opsin give insights into the mechanism of conversion of the molecular properties from invertebrate to vertebrate visual pigments.

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  • Cricket body size is altered by systemic RNAi against insulin signaling components and epidermal growth factor receptor 査読

    Noha Dabour, Tetsuya Bando, Taro Nakamura, Katsuyuki Miyawaki, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENT GROWTH & DIFFERENTIATION53 ( 7 ) 857 - 869   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    A long-standing problem of developmental biology is how body size is determined. In Drosophila melanogaster, the insulin/insulin-like growth factor (I/IGF) and target of rapamycin (TOR) signaling pathways play important roles in this process. However, the detailed mechanisms by which insect body growth is regulated are not known. Therefore, we have attempted to utilize systemic nymphal RNA interference (nyRNAi) to knockdown expression of insulin signaling components including Insulin receptor (InR), Insulin receptor substrate (chico), Phosphatase and tensin homologue (Pten), Target of rapamycin (Tor), RPS6-p70-protein kinase (S6k), Forkhead box O (FoxO) and Epidermal growth factor receptor (Egfr) and observed the effects on body size in the Gryllus bimaculatus cricket. We found that crickets treated with double-stranded RNA (dsRNA) against Gryllus InR, chico, Tor, S6k and Egfr displayed smaller body sizes, while Gryllus FoxO nyRNAi-ed crickets exhibited larger than normal body sizes. Furthermore, RNAi against Gryllus chico and Tor displayed slow growth and RNAi against Gryllus chico displayed longer lifespan than control crickets. Since no significant difference in ability of food uptake was observed between the Gryllus chico(nyRNAi) nymphs and controls, we conclude that the adult cricket body size can be altered by knockdown of expressions of Gryllus InR, chico, Tor, S6k, FoxO and Egfr by systemic RNAi. Our results suggest that the cricket is a promising model to study mechanisms underlying controls of body size and life span with RNAi methods.

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  • Ancestral functions of Delta/Notch signaling in the formation of body and leg segments in the cricket Gryllus bimaculatus 査読

    Taro Mito, Yohei Shinmyo, Kazuki Kurita, Taro Nakamura, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENT138 ( 17 ) 3823 - 3833   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Delta/Notch signaling controls a wide spectrum of developmental processes, including body and leg segmentation in arthropods. The various functions of Delta/Notch signaling vary among species. For instance, in Cupiennius spiders, Delta/Notch signaling is essential for body and leg segmentation, whereas in Drosophila fruit flies it is involved in leg segmentation but not body segmentation. Therefore, to gain further insight into the functional evolution of Delta/Notch signaling in arthropod body and leg segmentation, we analyzed the function of the Delta (Gb'Delta) and Notch (Gb'Notch) genes in the hemimetabolous, intermediate-germ cricket Gryllus bimaculatus. We found that Gb'Delta and Gb'Notch were expressed in developing legs, and that RNAi silencing of Gb'Notch resulted in a marked reduction in leg length with a loss of joints. Our results suggest that the role of Notch signaling in leg segmentation is conserved in hemimetabolous insects. Furthermore, we found that Gb'Delta was expressed transiently in the posterior growth zone of the germband and in segmental stripes earlier than the appearance of wingless segmental stripes, whereas Gb'Notch was uniformly expressed in early germbands. RNAi knockdown of Gb'Delta or Gb'Notch expression resulted in malformation in body segments and a loss of posterior segments, the latter probably due to a defect in posterior growth. Therefore, in the cricket, Delta/Notch signaling might be required for proper morphogenesis of body segments and posterior elongation, but not for specification of segment boundaries.

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  • Lowfat, a Mammalian Lix1 Homologue, Regulates Leg Size and Growth Under the Dachsous/Fat Signaling Pathway During Tissue Regeneration 査読

    Tetsuya Bando, Yoshimasa Hamada, Kazuki Kurita, Taro Nakamura, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENTAL DYNAMICS240 ( 6 ) 1440 - 1453   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    In the cricket Gryllus bimaculatus, missing distal parts of amputated legs are regenerated from blastemas based on positional information. The Dachsous/Fat (Ds/Ft) signaling pathway regulates blastema cell proliferation and positional information along the longitudinal axis during leg regeneration. Herein, we show that the Gryllus homologue of Lowfat (Gb'Lft), which modulates Ds/Ft signaling in Drosophila, is involved in leg regeneration. Gb'lft is expressed in regenerating legs, and RNAi against Gb'lft (Gb'lft RNAi) suppressed blastema cell hyperproliferation caused by Gb'ft(RNAi) or Gb'ds(RNAi) but enhanced that caused by Gb'kibra(RNAi) or Gb'warts(RNAi). In Gb'lft RNAi nymphs, missing parts of amputated legs were regenerated, but the length of the regenerated legs was shortened depending on the position of the amputation. Both normal and reversed intercalary regeneration occurred in Gb'lft(RNAi) nymphs, suggesting that Gb'Lft is involved in blastema cell proliferation and longitudinal leg regeneration under the Ds/Ft signaling pathway, but it is not required for intercalary regeneration. Developmental Dynamics 240:1440-1453, 2011. (C) 2011 Wiley-Liss, Inc.

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  • Regulation of Leg Size and Shape: Involvement of the Dachsous-Fat Signaling Pathway 査読

    Tetsuya Bando, Taro Mito, Taro Nakamura, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENTAL DYNAMICS240 ( 5 ) 1028 - 1041   2011年5月

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    記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    How limb size and shape is regulated is a long-standing question in developmental and regeneration biology. Recently, the protocadherin Dachsous-Fat (Ds-Ft) signaling pathway has been found to be essential for wing development of the fly and leg regeneration of the cricket. The Ds-Ft signaling pathway is linked to the Warts-Hippo (Wts-Hpo) signaling pathway, leading to cell proliferation. Several lines of evidence have suggested that the Wts-Hpo signaling pathway is involved in the control of organ size, and that this pathway is regulated by Ds-Ft and Merlin-Expanded, which are linked to morphogens such as decapentaplegic/bone morphogenic protein, Wingless/Wnt, and epidermal growth factor. Here we review recent progress in elucidating mechanisms controlling leg size and shape in insects and vertebrates, focusing on the Ds-Ft signaling pathway. We also introduce a working model, Ds-Ft steepness model, to explain how steepness of the Ds-Ft gradient controls leg size along the proximodistal axis. Developmental Dynamics 240:1028-1041, 2011. (C) 2011 Wiley-Liss, Inc.

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  • Autotaxin Is Required for the Cranial Neural Tube Closure and Establishment of the Midbrain-Hindbrain Boundary During Mouse Development 査読

    Seiichi Koike, Yoshifumi Yutoh, Kazuko Keino-Masu, Sumihare Noji, Masayuki Masu, Hideyo Ohuchi

    DEVELOPMENTAL DYNAMICS240 ( 2 ) 413 - 421   2011年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Autotaxin (ATX) is a lysophospholipid-generating exoenzyme expressed in embryonic and adult neural tissues. We previously showed that ATX is expressed in the neural organizing centers, anterior head process, and midbrain-hindbrain boundary (MHB). To elucidate the role of ATX during neural development, here we examined the neural phenotypes of ATX-deficient mice. Expression analysis of neural marker genes revealed that lateral expansion of the rostral forebrain is reduced and establishment of the MHB is compromised as early as the late headfold stage in ATX mutant embryos. Moreover, ATX mutant embryos fail to complete cranial neural tube closure. These results indicate that ATX is essential for cranial neurulation and MHB establishment. Developmental Dynamics 240:413-421, 2011. (C) 2011 Wiley-Liss, Inc.

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  • Autotaxin is required for the cranial neural tube closure and establishment of the midbrain-hindbrain boundary during mouse development 査読

    Seiichi Koike, Yoshifumi Yutoh, Kazuko Keino-Masu, Sumihare Noji, Masayuki Masu, Hideyo Ohuchi

    Developmental Dynamics240 ( 2 ) 413 - 421   2011年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Autotaxin (ATX) is a lysophospholipid-generating exoenzyme expressed in embryonic and adult neural tissues. We previously showed that ATX is expressed in the neural organizing centers, anterior head process, and midbrain-hindbrain boundary (MHB). To elucidate the role of ATX during neural development, here we examined the neural phenotypes of ATX-deficient mice. Expression analysis of neural marker genes revealed that lateral expansion of the rostral forebrain is reduced and establishment of the MHB is compromised as early as the late headfold stage in ATX mutant embryos. Moreover, ATX mutant embryos fail to complete cranial neural tube closure. These results indicate that ATX is essential for cranial neurulation and MHB establishment. © 2011 Wiley-Liss, Inc.

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  • Atelocollagen-mediated systemic administration of myostatin-targeting siRNA improves muscular atrophy in caveolin-3-deficient mice 査読

    Emi Kawakami, Nao Kinouchi, Taro Adachi, Yutaka Ohsawa, Naozumi Ishimaru, Hideyo Ohuchi, Yoshihide Sunada, Yoshio Hayashi, Eiji Tanaka, Sumihare Noji

    DEVELOPMENT GROWTH & DIFFERENTIATION53 ( 1 ) 48 - 54   2011年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL PUBLISHING, INC  

    Small interfering RNA (siRNA)-mediated silencing of gene expression is rapidly becoming a powerful tool for molecular therapy. However, the rapid degradation of siRNAs and their limited duration of activity require efficient delivery methods. Atelocollagen (ATCOL)-mediated administration of siRNAs is a promising approach to disease treatment, including muscular atrophy. Herein, we report that ATCOL-mediated systemic administration of a myostatin-targeting siRNA into a caveolin-3-deficient mouse model of limb-girdle muscular dystrophy 1C (LGMD1C) induced a marked increase in muscle mass and a significant recovery of contractile force. These results provide evidence that ATCOL-mediated systemic administration of siRNAs may be a powerful therapeutic tool for disease treatment, including muscular atrophy.

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  • Opn5 is a UV-sensitive bistable pigment that couples with Gi subtype of G protein 査読

    Takahiro Yamashita, Hideyo Ohuchi, Sayuri Tomonari, Keiko Ikeda, Kazumi Sakai, Yoshinori Shichida

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA107 ( 51 ) 22084 - 22089   2010年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATL ACAD SCIENCES  

    Opn5 (neuropsin) belongs to an independent group separated from the other six groups in the phylogenetic tree of opsins, for which little information of absorption characteristics and molecular properties of the members is available. Here we show that the chicken Opn5 (cOpn5m) is a UV-sensitive bistable pigment that couples with Gi subtype of G protein. The recombinant expression of cOpn5m in HEK 293s cells followed by the addition of 11-cis- and all-trans-retinal produced UV light-absorbing and visible light-absorbing forms, respectively. These forms were interconvertible by UV and visible light irradiations, respectively, indicating that cOpn5m is a bistable pigment. The absorption maxima of these forms were estimated to be 360 and 474 nm, respectively. The GTP gamma S binding assay clearly showed that the visible light-absorbing form having all-trans-retinal activates Gi type of G protein, whereas no Gt or Gq activation ability was observed. Immunohistochemical studies using an antibody against cOpn5m clearly showed that this pigment is localized within some types of amacrine cells and some cells in the ganglion cell layer of the retinas, the vast majority of cells in the pineal gland and serotonin-positive cells in the paraventricular organ. Because cOpn5m is the only UV-sensitive opsin among the opsins found so far in chicken, this study provides the molecular basis for UV reception in chicken.

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  • Delivery of small interfering RNA with a synthetic collagen poly(Pro-Hyp-Gly) for gene silencing in vitro and in vivo 査読

    Taro Adachi, Emi Kawakami, Naozumi Ishimaru, Takahiro Ochiya, Yoshio Hayashi, Hideyo Ohuchi, Masao Tanihara, Eiji Tanaka, Sumihare Noji

    DEVELOPMENT GROWTH & DIFFERENTIATION52 ( 8 ) 693 - 699   2010年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Silencing gene expression by small interfering RNAs (siRNAs) has become a powerful tool for the genetic analysis of many animals. However, the rapid degradation of siRNA and the limited duration of its action in vivo have called for an efficient delivery technology. Here, we describe that siRNA complexed with a synthetic collagen poly(Pro-Hyp-Gly) (SYCOL) is resistant to nucleases and is efficiently transferred into cells in vitro and in vivo, thereby allowing long-term gene silencing in vivo. We found that the SYCOL-mediated local application of siRNA targeting myostatin, coding a negative regulator of skeletal muscle growth, in mouse skeletal muscles, caused a marked increase in the muscle mass within a few weeks after application. Furthermore, in vivo administration of an anti-luciferase siRNA/SYCOL complex partially reduced luciferase expression in xenografted tumors in vivo. These results indicate a SYCOL-based non-viral delivery method could be a reliable simple approach to knockdown gene expression by RNAi in vivo as well as in vitro.

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  • Autotaxin Controls Caudal Diencephalon-Mesencephalon Development in the Chick 査読

    Hideyo Ohuchi, Hitomi Fukui, Akane Matsuyo, Sayuri Tomonari, Masayuki Tanaka, Hiroyuki Arai, Sumihare Noji, Junken Aoki

    DEVELOPMENTAL DYNAMICS239 ( 10 ) 2647 - 2658   2010年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The diencephalon is the embryonic anlagen of the higher integration centers of the brain. Recent studies have elucidated how the cells in the rostral diencephalon acquire their regional identities. However, the understanding of the mechanisms under which the caudal diencephalon is formed is still limited. Here we focus on the role of Autotaxin (ATX), a lysophospholipid-generating exoenzyme, whose mRNA is detected in the caudal diencephalon. RNA interference against ATX altered the expression pattern of Pax6-regualted genes, Tcf4, Lim1, and En1, implying that ATX is required for the maintenance of the regional identity of the caudal diencephalon and the diencephalon-mesencephalon boundary (DMB). Furthermore, ATX-RNAi inhibited neuroepithelial cell proliferation on both sides of the DMB. We propose a dual role of ATX in chick brain development, in which ATX not only contributes to the formation of caudal diencephalon as a short-range signal, but also regulates the growth of mesencephalon as a long-range signal. Developmental Dynamics 239:2647-2658, 2010. (C) 2010 Wiley-Liss, Inc.

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  • Imaging of Transgenic Cricket Embryos Reveals Cell Movements Consistent with a Syncytial Patterning Mechanism 査読

    Taro Nakamura, Masato Yoshizaki, Shotaro Ogawa, Haruko Okamoto, Yohei Shinmyo, Tetsuya Bando, Hideyo Ohuchi, Sumihare Noji, Taro Mito

    CURRENT BIOLOGY20 ( 18 ) 1641 - 1647   2010年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    The mode of insect embryogenesis varies among species, reflecting adaptations to different lite history strategies [1, 2]. In holometabolous insects, which include the model systems, such as the fruit fly and the red flour beetle, a large proportion of the blastoderm produces an embryo, whereas hemimetabolous embryos generally arise from a small region of the blastoderm [3]. Despite their importance in evolutionary studies, information of early developmental dynamics of hemimetabolous insects remains limited. Here, to clarify how maternal and gap gene products act in patterning the embryo of basal hemimetabolous insects, we analyzed the dynamic segmentation process in transgenic embryos of an intermediate-germ insect species, the cricket, Gryllus bimaculatus. Our data based on live imaging of fluorescently labeled embryonic cells and nuclei suggest that the positional specification of the cellular blastoderm may be established in the syncytiurn, where maternally derived gradients could act fundamentally in a way that is similar to that of Drosophila, namely throughout the egg. Then, the blastoderm cells move dynamically, retaining their positional information to form the posteriorly localized germ anlage. Furthermore, we find that the anterior head region of the cricket embryo is specified by orthodenticle in a cellular environment earlier than the gnathal and thoracic regions. Our findings imply that the syncytial mode of the early segmentation in long-germ insects evolved from a dynamic syncytial-to-cellular mode found in the present study, accompanied by a heterochronic shift of gap gene action.

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  • Loss-of-Function Analyses of the Fragile X-Related and Dopamine Receptor Genes by RNA Interference in the Cricket Gryllus bimaculatus 査読

    Aska Hamada, Katsuyuki Miyawaki, Eri Honda-sumi, Kenji Tomioka, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENTAL DYNAMICS238 ( 8 ) 2025 - 2033   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    In order to explore a possibility that the cricket Gryllus bimaculatus would be a useful model to unveil molecular mechanisms of human diseases, we performed loss-of-function analyses of Gryllus genes homologous to human genes that are responsible for human disorders, fragile X mental retardation 1 (fmr1) and Dopamine receptor (DopR). We cloned cDNAs of their Gryllus homologues, Gb'fmr1, Gb'DopRI, and Gb'DopRII, and analyzed their functions with use of nymphal RNA interference (RNAi). For Gb'fmr1, three major phenotypes were observed: (1) abnormal wing postures, (2) abnormal calling song, and (3) loss of the circadian locomotor rhythm, while for Gb'DopRI, defects of wing posture and morphology were found. These results indicate that the cricket has the potential to become a novel model system to explore human neuronal pathogenic mechanisms and to screen therapeutic drugs by RNAi. Developmental Dynamics 238. 2025-2033, 2009. (C) 2009 Wiley-Liss, Inc.

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  • Regulation of leg size and shape by the Dachsous/Fat signalling pathway during regeneration 査読

    Tetsuya Bando, Taro Mito, Yuko Maeda, Taro Nakamura, Fumiaki Ito, Takahito Watanabe, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENT136 ( 13 ) 2235 - 2245   2009年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    An amputated cricket leg regenerates all missing parts with normal size and shape, indicating that regenerating blastemal cells are aware of both their position and the normal size of the leg. However, the molecular mechanisms regulating this process remain elusive. Here, we use a cricket model to show that the Dachsous/Fat (Ds/Ft) signalling pathway is essential for leg regeneration. We found that knockdown of ft or ds transcripts by regeneration-dependent RNA interference (rdRNAi) suppressed proliferation of the regenerating cells along the proximodistal (PD) axis concomitantly with remodelling of the pre-existing stump, making the regenerated legs shorter than normal. By contrast, knockdown of the expanded (ex) or Merlin (Mer) transcripts induced over-proliferation of the regenerating cells, making the regenerated legs longer. These results are consistent with those obtained using rdRNAi during intercalary regeneration induced by leg transplantation. We present a model to explain our results in which the steepness of the Ds/Ft gradient controls growth along the PD axis of the regenerating leg.

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  • Subtype-specific expression of Fgf19 during horizontal cell development of the chicken retina 査読

    Mayumi Okamoto, Takaaki Bito, Sumihare Noji, Hideyo Ohuchi

    GENE EXPRESSION PATTERNS9 ( 5 ) 306 - 313   2009年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The mechanisms underlying retinal cell diversification are crucial to proper neural development. Fibroblast growth factor 19 (Fgf19) is expressed by developing horizontal cells (HCs) in the chicken retina. Although there are two major HC subtypes, axon-bearing and axon-less, the precise subtype expressing Fgf19 remains uncertain. Here we characterize Fgf19-expressing cells by co-labeling with antibodies against Lim1 (LIM homeodomain 1, or Lhx1), Islet1, and Prox1 (prospero-related homeobox 1) which are axon-bearing HC, axon-less HC, and pan-HC markers, respectively. We found that a subset of Fgr19-expressing cells was positive for Prox1 and Lim1 in the vitread neuroepithelium at embryonic day 4 (E4). By E9, the majority of Fgf19-expressing cells became positive for Prox1 and Lim1 prior to arrival at the prospective HC layer. In contrast, Fgf19-expressing cells did not overlap with the Islet1-positive population at any stage examined. These results suggest that Fgf19 is expressed by the early migratory horizontal precursors, and later by the presumptive axon-bearing HCs. (C) 2009 Elsevier B.V. All rights reserved.

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  • Expression Patterns of the Lysophospholipid Receptor Genes During Mouse Early Development 査読

    Hideyo Ohuchi, Aska Hamada, Hironao Matsuda, Akira Takagi, Masayuki Tanaka, Junken Aoki, Hiroyuki Arai, Sumihare Noji

    DEVELOPMENTAL DYNAMICS237 ( 11 ) 3280 - 3294   2008年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Lysophospholipids (LPs) such as lysophosphatidic acid (LPA) and sphingosine-1-phosphate (SIP) are known to mediate various biological responses, including cell proliferation, migration, and differentiation. To better understand the role of these lipids in mammalian early development, we applied whole-mount in situ hybridization techniques to E8.5 to E12.5 mouse embryos. We determined the expression patterns of the following LP receptor genes, which belong to the G protein-coupled receptor (GPCR) family: EDG1 to EDG8 (S1P(1) to S1P(5) and LPA(1) to LPA(3)), LPA(4) (GPR23 /P2Y9), and LPA(5) (GPR92). We found that the S1P/LPA receptor genes exhibit overlapping expression patterns in a variety of organ primordia, including the developing brain and cardiovascular system, presomitic mesoderm and somites, branchial arches, and limb buds. These results suggest that multiple receptor systems for LPA/S1P lysophospholipids may be functioning during organogenesis. Developmental Dynamics 237.3280-3294, 2008. (C) 2008 Wiley-Liss, Inc.

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  • Atelocollagen-mediated local and systemic applications of myostatin-targeting siRNA increase skeletal muscle mass.

    Kinouchi N, Ohsawa Y, Ishimaru N, Ohuchi H, Sunada Y, Hayashi Y, Tanimoto Y, Moriyama K, Noji S

    Gene Therapy15 ( 15 ) 1126 - 1130   2008年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    RNA interference (RNAi) offers a novel therapeutic strategy based on the highly specific and efficient silencing of a target gene. Since it relies on small interfering RNAs (siRNAs), a major issue is the delivery of therapeutically active siRNAs into the target tissue/target cells in vivo. For safety reasons, strategies based on vector delivery may be of only limited clinical use. The more desirable approach is to directly apply active siRNAs in vivo. Here, we report the effectiveness of in vivo siRNA delivery into skeletal muscles of normal or diseased mice through nanoparticle formation of chemically unmodified siRNAs with atelocollagen (ATCOL). ATCOL-mediated local application of siRNA targeting myostatin, a negative regulator of skeletal muscle growth, in mouse skeletal muscles or intravenously, caused a marked increase in the muscle mass within a few weeks after application. These results imply that ATCOL-mediated application of siRNAs is a powerful tool for future therapeutic use for diseases including muscular atrophy.

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  • Expression patterns of the opsin 5-related genes in the developing chicken retina 査読

    Sayuri Tomonari, Kyoichi Migita, Akira Takagi, Sumihare Noji, Hideyo Ohuchi

    DEVELOPMENTAL DYNAMICS237 ( 7 ) 1910 - 1922   2008年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The opsin gene family encodes G protein-coupled seven-transmembrane proteins that bind to a retinaldehyde chromophore for photoreception. It has been reported that opsin 5 is expressed in mammalian neural tissue, but its function has been elusive. As a first step to understand the function for opsin 5 in the developing eye, we searched for chicken opsin 5-related genes in the genome by a bioinformatic approach and isolated opsin 5 cDNA fragments from the embryonic retina by RT-PCR. We found that there are three opsin 5-related genes, designated cOpn5m (chicken opsin 5, mammalian type), cOpn5L1 (chicken opsin 5-like 1), and cOpn5L2 (chicken opsin 5-like 2), in the chicken genome. Quantitative PCR analysis has revealed that cOpn5m is the most abundant in the developing and early posthatching neural retina. In situ hybridization analysis has shown that cOpn5m is specifically expressed in subsets of differentiating ganglion cells and amacrine cells. These results suggest that the mammalian type opsin 5 may contribute to the development of these retinal cells in the chicken.

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  • EGFR signaling is required for re-establishing the proximodistal axis during distal leg regeneration in the cricket Gryllus bimaculatus nymph 査読

    Taro Nakamura, Taro Mito, Katsuyuki Miyawaki, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENTAL BIOLOGY319 ( 1 ) 46 - 55   2008年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Nymphs of hemimetabolous insects, such as cockroaches and crickets, possess functional legs with a remarkable capacity for epimorphic regeneration. In this study, we have focused on the role of epidermal growth factor receptor (EGFR) signaling in regeneration of a nymphal leg in the cricket Gryllus bimaculatus. We performed loss-of-function analyses with a Gryllus Egfr homolog (Gb'Egfr) and nymphal RNA interference (RNAi). After injection of double-stranded RNA for Gb'Egfr in the body cavity of the third instar cricket nymph, amputation of the leg at the distal tibia resulted in defects of normal distal regeneration. The regenerated leg lacked the distal tarsus and pretarsus. This result indicates that EGFR signaling is required for distal leg patterning in regeneration during the nymphal stage of the cricket. Furthermore, we demonstrated that EGFR signaling acts downstream of the canonical Wnt/Wg signaling and regulates appendage proximodistal (PD) patterning genes aristaless and dachshund during regeneration. Our results suggest that EGFR signaling influences positional information along the PD axis in distal leg patterning of insects, regardless of the leg formation mode. (C) 2008 Elsevier Inc. All rights reserved.

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  • Dynamic expression patterns of vasa during embryogenesis in the cricket Gryllus bimaculatus 査読

    Taro Mito, Taro Nakamura, Isao Sarashina, Chun-che Chang, Shotaro Ogawa, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENT GENES AND EVOLUTION218 ( 7 ) 381 - 387   2008年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    The specification of germ cells during embryogenesis is an important issue in the development of metazoans. In insects, the mode of germ cell specification appears to be highly variable among species and molecular data are not sufficient to provide an evolutionary perspective to this issue. Expression of vasa can be used as a germ line marker. Here, we report the isolation of a vasa-like gene in a hemimetabolous insect, the cricket Gryllus bimaculatus (Gb'vas), and its expression patterns during oogenesis and embryogenesis. Gb'vas is preferentially expressed in the germarium and the expression of Gb'vas is detectable throughout vitellogenesis including mature eggs subjected to oviposition, suggesting that Gb'vas is maternally contributed to the cricket eggs. The zygotic expression of Gb'vas appears to start at the mid blastoderm stage in the posterior region of the egg, expanding in a developing germ anlage. In early germbands, an intense expression of Gb'vas is restricted to the posterior end. In later embryos, Gb'vas expression extends over the whole body and then distinctly localized to the embryonic gonad at the stage immediately before hatching. These results suggest that, in the cricket, germ cells are specified early in development at the posterior end of an early germband, as proposed by Heymons (1895) based on cytological criteria.

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  • Introduction of silencing-inducing transgene against Fgf19 does not affect expression of Tbx5 and beta 3-tubulin in the developing chicken retina 査読

    Mayumi Okamoto, Sayuri Tomonari, Yuki Naito, Kaoru Saigo, Sumihare Noji, Kumiko Ui-Tei, Hideyo Ohuchi

    DEVELOPMENT GROWTH & DIFFERENTIATION50 ( 3 ) 159 - 168   2008年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Fgf19 is known to be expressed in the developing chicken eye but its functions during retinal development have remained elusive. Since Fgf19 is expressed in the dorsal portion of the optic cup, it is intriguing to know whether FGF19 is required for expression of dorso-ventral morphogenetic genes in the eye. To clarify this, expression patterns of Tbx5 and Vax were examined in the developing eye after in ovo RNA interference targeted against Fgf19. Quantitative polymerase chain reaction (PCR) analysis showed that the short-hairpin RNAs (shRNAs) targeted against Fgf19 could reduce its expression in the eye to less than 50% of a relative amount of mRNA, compared with contralateral or untreated control eyes. However, no obvious alteration in expression domains of Tbx5 or Vax was observed. Misexpression of Tbx5 or Tbx5-RNAi did not alter the Fgf19 expression either. Furthermore, although Fgf19 is expressed in the central retina before neurogenesis occurs, beta 3-tubulin, a marker for early retinal differentiation was still detected in the central retina after knockdown of Fgf19. Thus, knockdown of Fgf19 supports no obvious regulations between Fgf19 and Tbx5, or exhibits no phenotypes that perturb early retinal differentiation.

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  • Divergent and conserved roles of extradenticle in body segmentation and appendage formation, respectively, in the cricket Gryllus bimaculatus 査読

    Taro Mito, Monica Ronco, Tomohiro Uda, Taro Nakamura, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENTAL BIOLOGY313 ( 1 ) 67 - 79   2008年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The cricket Gryllus bimaculatus is a typical hemimetabolous intemediate germ insect, in which the processes of segmentation and appendage formation differ from those in Drosophila, a holometabolous long germ insect. In order to compare their developmental mechanisms, we have focused on Gryllus orthologs of the Drosophila developmental regulatory genes and studied their functions. Here, we report a functional analysis of the Gryllus ortholog of extradenticle (Gb'exd) using embryonic and parental RNA interference (RNAi) techniques. We found the following: (1) RNAi suppression of Gb'exd results in the deletion or fusion of body segments. Especially the head was often very severely affected. This gap-like phenotype may be related to reduced expression of the gap genes hunchback and Kruppel in early RNAi germbands. (2) In the appendages, several segments (podomeres) were fused. (3) Head appendages including the antenna were transformed to a leg-like structure consisting of at least one proximal podomere as well as several tarsomeres. The defects in appendages are reminiscent of the phenotype caused by large exd clones in Drosophila antennal discs. These findings led us to the conclusion that (1) Gb'exd is required for segment patterning in the gnathal to abdominal region, acting in a gap gene-like manner in the anterior region. (2) Gb'exd plays important roles in formation of the appendages and the determination of their identities, acting as a regulatory switch that chooses between the fates of head appendages versus the appendage ground state. Although functions of Gbexd in appendage patterning appear fundamentally conserved between Gryllus and Drosophila, its role in body segmentation may differ from that of Drosophila exd. (c) 2007 Elsevier Inc. All rights reserved.

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  • Film tomography as a tool for three-dimensional image construction and gene expression studies 査読

    Masayuki Mitsumori, Taro Adachi, Kazuho Takayanagi, Taro Mito, Hideyo Ohuchi, Shouichi Kimura, Mitsunori Kokubo, Toshiro Higuchi, Sumihare Noji

    DEVELOPMENT GROWTH & DIFFERENTIATION49 ( 7 ) 583 - 589   2007年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    In order to observe three-dimensional (3D) expression patterns of genes in whole animals, whole organs, or whole tissues, in situ hybridization (ISH) of many sections must be carried out and then used to construct a 3D image. For this purpose, we have developed an automatic microtome to prepare tissue sections with an adhesive film. We used commercially available film suitable for sectioning and ISH. We constructed a microtome and, after adherence of the film to a paraffin-embedded tissue block, cut the block with a blade to prepare sections on film. Then, the sections-on-film were automatically set in a plastic frame that was the same size as a conventional glass slide. With this automatic microtome, tissue sections can be made for ISH or immunohistochemistry in addition to conventional hematoxylin and eosin staining without specific training. We demonstrate that we can construct 3D images of gene expression patterns obtained by ISH on sections prepared with this automatic microtome. We have designated this method as 'Film Tomography (FITO)'.

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  • Expression pattern of the melanopsin-like (cOpn4m) and VA opsin-like genes in the developing chicken retina and neural tissues 査読

    Sayuri Tomonari, Akira Takagi, Sumittare Noji, Hideyo Ohuchi

    GENE EXPRESSION PATTERNS7 ( 7 ) 746 - 753   2007年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    We examined the expression pattern of melanopsin-like (cOpn4m) and VA opsin-like (cVAL) genes during chicken development. Two types of cOpn4m transcripts, distinct in their carboxyl terminals were found, as is the case for the chicken melanopsin (cOpn4) reported previously. The expression of cOpn4m was restricted to the developing retina, specifically to a subset of developing amacrine cells from embryonic day 10. VA opsin is one of the non-canonical opsins, reported to exist in fish so far. In this study, an aberrant type of VA opsin-like (cVAL) cDNA was isolated from chicken embryonic neural tissues. The expression of cVAL was observed in the ventral region of the developing brain and neural tube; however, specific signals for cVAL could not be detected in the developing retina. These results indicate that the additional melanopsin in avian identifies a subset of developing amacrine cells in the retina and that the aberrant transcript of the VA opsin-like gene are present during neural tube development in the chicken. (c) 2007 Elsevier B.V. All rights reserved.

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  • Diversified expression patterns of autotaxin, a gene for phospholipid-generating enzyme during mouse and chicken development (vol 236, pg 1134, 2007) 査読

    Ohuchi Hideyo, Hayashibara Yasunori, Matsuda Hironao, Onoi Motoyoshi, Mitsumori Masayuki, Tanaka Masayuki, Aoki Junken, Arai Hiroyuki, Noji Sumihare

    DEVELOPMENTAL DYNAMICS236 ( 5 ) 1376   2007年5月

  • Diversified expression patterns of autotaxin, a gene for phospholipid-generating enzyme during mouse and chicken development 査読

    Hideyo Ohuchi, Yasunori Hayashibaral, Hironao Matsuda, Motoyoshi Onoi, Masayuki Mitsumori, Masayuki Tanaka, Junken Aoki, Hiroyuki Arai, Sumihare Noji

    DEVELOPMENTAL DYNAMICS236 ( 4 ) 1134 - 1143   2007年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Autotaxin (ATX), or nucleotide pyrophosphatase-phosphodiesterase 2, is a secreted lysophospholipase D that generates bioactive phospholipids that act on G protein-coupled receptors. Here we show the expression patterns of the ATX gene in mouse and chicken embryos. ATX has a dynamic spatial and temporal expression pattern in both species and the expression domains during neural development are quite distinct from each other. Murine ATX (mATX) is expressed immediately rostral to the midbrain-hindbrain boundary, whereas chicken ATX (cATX) is expressed in the diencephalon and later in the parencephalon-synencephalon boundary. In the neural tube, cATX is expressed in the alar plate in contrast to mATX in the floor plate. ATX is also expressed in the hindbrain and various organ primordia such as face anlagen and skin appendages of the mouse and chicken. These results suggest conserved and non-conserved roles for ATX during neural development and organogenesis in these species.

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  • even-skipped has gap-like, pair-rule-like, and segmental functions in the cricket Gryllus bimaculatus, a basal, intermediate germ insect (Orthoptera) 査読

    Taro Mito, Chiharu Kobayashi, Isao Sarashina, Hongjie Zhang, Wakako Shinahara, Katsuyuki Miyawaki, Yohei Shinmyo, Hideyo Ohuchi, Sumihare Noji

    Developmental Biology303 ( 1 ) 202 - 213   2007年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Academic Press Inc.  

    Developmental mechanisms of segmentation appear to be varied among insects in spite of their conserved body plan. Although the expression patterns of the segment polarity genes in all insects examined imply well conserved function of this class of genes, expression patterns and function of the pair-rule genes tend to exhibit diversity. To gain further insights into the evolution of the segmentation process and the role of pair-rule genes, we have examined expression and function of an ortholog of the Drosophila pair-rule gene even-skipped (eve) in a phylogenetically basal insect, Gryllus bimaculatus (Orthoptera, intermediate germ cricket). We find that Gryllus eve (Gb'eve) is expressed as stripes in each of the prospective gnathal, thoracic, and abdominal segments and as a broad domain in the posterior growth zone. Dynamics of stripe formation vary among Gb'eve stripes, representing one of the three modes, the segmental, incomplete pair-rule, and complete pair-rule mode. Furthermore, we find that RNAi suppression of Gb'eve results in segmentation defects in both anterior and posterior regions of the embryo. Mild depletion of Gb'eve shows a pair-rule-like defect in anterior segments, while stronger depletion causes a gap-like defect showing deletion of anterior and posterior segments. These results suggest that Gb'eve acts as a pair-rule gene at least during anterior segmentation and also has segmental and gap-like functions. Additionally, Gb'eve may be involved in the regulation of hunchback and Krüppel expression. Comparisons with eve functions in other species suggest that the Gb'eve function may represent an intermediate state of the evolution of pair-rule patterning by eve in insects. © 2006 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.ydbio.2006.11.003

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  • Involvement of canonical Wnt/Wingless signaling in the determination of the positional values within the leg segment of the cricket Gryllus bimaculatus 査読

    Taro Nakamura, Taro Mito, Yoshihisa Tanaka, Tetsuya Bando, Hideyo Ohuchi, Sumihare Noji

    DEVELOPMENT GROWTH & DIFFERENTIATION49 ( 2 ) 79 - 88   2007年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    The cricket Gryllus bimaculatus is a hemimetabolous insect whose nymphs posses the ability to regenerate amputated legs. Previously, we showed that Gryllus orthologues of Drosophila hedgehog (Gb'hh), wingless (Gb'wg) and decapentaplegic (Gb'dpp) are expressed during leg regeneration and play essential roles in the establishment of the proximal-distal axis. Here, we examined their roles during intercalary regeneration: when a distally amputated tibia with disparate positional values is placed next to a proximally amputated host, intercalary growth occurs in order to regenerate the missing part. In this process, we examined expression patterns of Gb'hh and Gb'wg. We found that expressions of Gb'hh and Gb'wg were induced in a regenerate and the host proximal to the amputated region, but not in the grafted donor distal to the regenerate. This directional induction occurs even in the reversed intercalation. Because these results are consistent with a distal-to-proximal respecification of the regenerate, Gb'wg may be involved in the re-establishment of the positional values in the regenerate. Furthermore, we found that no regeneration occurs when Gb'armadillo (the orthologue of beta-catenin) was knocked down by RNA interference. These results indicate that the canonical Wnt/Wingless signaling pathway is involved in the process of leg regeneration and determination of positional information in the leg segment.

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  • Characterization of follistatin-related gene as a negative regulatory factor for activin family members during mouse heart development 査読

    Yuka Takehara-Kasamatsu, Kunihiro Tsuchida, Masashi Nakatani, Tatsuya Murakami, Akira Kurisaki, Osamu Hashimoto, Hideyo Ohuchi, Hitomi Kurose, Kazuhiro Mori, Shoji Kagami, Sumihare Noji, Hiromu Sugino

    Journal of Medical Investigation54 ( 3-4 ) 276 - 288   2007年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:3-4  

    Follistatin-related gene (FLRG) encodes a secretory glycoprotein that has characteristic cysteine-rich follistatin domains. FLRG protein binds to and neutralizes several transforming growth factor-β (TGF-β) superfamily members, including myostatin (MSTN), which is a potent negative regulator of skeletal muscle mass. We have previously reported that FLRG was abundantly expressed in fetal and adult mouse heart. In this study, we analyzed the expression of FLRG mRNA during mouse heart development. FLRG mRNA was continuously expressed in the embryonic heart, whereas it was very low in skeletal muscles. By contrast, MSTN mRNA was highly expressed in embryonic skeletal muscles, whereas the expression of MSTN mRNA was rather low in the heart. In situ hybridization and immunohistochemical analysis revealed that FLRG expressed in smooth muscle of the aorta and pulmonary artery, valve leaflets of mitral and tricuspid valves, and cardiac muscles in the ventricle of mouse embryonic heart. However, MSTN was expressed in very limited areas, such as valve leaflets of pulmonary and aortic valves, the top of the ventricular and atrial septa. Interestingly, the expression of MSTN was complementary to that of FLRG, especially in the valvular apparatus. Biochemical analyses with surface plasmon resonance biosensor and reporter assays demonstrated that FLRG hardly dissociates from MSTN and activin once it bound to them, and efficiently inhibits these activities. Our results suggest that FLRG could function as a negative regulator of activin family members including MSTN during heart development.

    DOI: 10.2152/jmi.54.276

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  • Brachyenteron is necessary for morphogenesis of theposterior gut but not for anteroposterior axial elongationfrom the posterior growth zone in the intermediate-germbandcricket Gryllus bimaculatus 査読

    Yohei Shinmyo, Taro Mito, Tomohiro Uda, Taro Nakamura, Katsuyuki Miyawaki, Hideyo Ohuchi, Sumihare Noji

    Development133 ( 22 ) 4539 - 4547   2006年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In the long-germband insect Drosophila, all body segments and posterior terminal structures, including the posterior gut and anal pads, are specified at the blastoderm stage. In short- and intermediate-germband insects, however, posterior segments are sequentially produced from the posterior growth zone, a process resembling somitogenesis in vertebrates, and invagination of the posterior gut starts after anteroposterior (AP) axial elongation from the growth zone. The mechanisms underlying posterior segmentation and terminal patterning in these insects are poorly understood. In order to elucidate these mechanisms, we have investigated the roles of the Brachyurylbrachyenteron (Bra/byn) homolog in the intermediate-germ band cricket Gryllus bimaculatus. Loss-of-function analysis by RNA interference (RNAi) revealed that Gryllus byn (Gb'byn) is not required for AP axial elongation or normal segment formation, but is required for specification of the posterior gut. We also analyzed Gryllus caudal (Gb'cad) RNAi embryos using in situ hybridization with a Gb'byn probe, and found that Gb'cad is required for internalization of the posterior gut primordium, in addition to AP axial elongation. These results suggest that the functions of byn and cad in posterior terminal patterning are highly conserved in Gryllus and Drosophila despite their divergent posterior patterning. Moreover, because it is thought that the progressive growth of the AP axis from the growth zone, controlled by a genetic program involving Cdx/cad and Bra/byn, might be ancestral to bilaterians, our data suggest that the function of Bra/byn in this process might have been lost in insects.

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  • Krüppel acts as a gap gene regulating expression of hunchback and even-skipped in the intermediate germ cricket Gryllus bimaculatus. 査読

    Mito T, Okamoto H, Shinahara W, Shinmyo Y, Miyawaki K, Ohuchi H, Noji S

    Developmental biology294 ( 2 ) 471 - 481   2006年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.ydbio.2005.12.057

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  • Exogenous FGF10 can rescue an eye-open at birth phenotype of Fgf10-null mice by activating activin and TGF alpha-EGFR signaling 査読

    H Tao, K Ono, H Kurose, S Noji, H Ohuchi

    DEVELOPMENT GROWTH & DIFFERENTIATION48 ( 5 ) 339 - 346   2006年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    Mutant mice deficient in the fibroblast growth factor 10 (Fgf10) gene exhibit an eye-open phenotype at birth. It has previously been shown that FGF10 has a dual role in proliferation and migration during the early and later stages of eyelid development, respectively. To verify the role of FGF10 during eyelid closure, explant culture of Fgf10-null eyelid anlagen was performed, by which it was examined whether or not exogenous FGF10 could rescue the expression of activin beta B and transforming growth factor alpha, known to be required for eyelid closure. We found that the expression of these genes was markedly induced while that of Shh or Ptch1, Ptch2 was not. We also observed the distribution of filamentous actin (F-actin) after FGF10 application in the mutant eyelid explant, finding that the FGF10 protein induced F-actin accumulation. We further examined filopodia of the eyelid leading edge cells, finding the length of the filopodia was significantly reduced in the mutant. These results verify that FGF10 promotes eyelid closure through activating activin and TGF alpha-EGFR signaling.

    DOI: 10.1111/j.1440-169x.2006.00869.x

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  • Cessation of Fgf10 signaling, resulting in a defective dental epithelial stem cell compartment, leads to the transition from crown to root formation 査読

    T Yokohama-Tamaki, H Ohshima, N Fujiwara, Y Takada, Y Ichimori, S Wakisaka, H Ohuchi, H Harada

    DEVELOPMENT133 ( 7 ) 1359 - 1366   2006年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Mouse, rat and human molars begin to form root after the completion of crown formation. In these teeth, fibroblast growth factor (Fgf) 10 disappears in the transitional stage from crown formation to root. By contrast, rodent incisors and vole molars demonstrate continuous growth, owing to the formation and maintenance of a stem cell compartment by the constant expression of Fgf10. To clarify the relationship between root formation and disappearance of Fgf10, we carried out two experiments for the loss and gain of Fgf10 function. First, we examined postnatal growth in the incisors of Fgf10-deficient mice, which have the defect of a dental epithelial stem cell compartment referred to as 'apical bud', after implantation under the kidney capsule. The growth at the labial side in the mutant mice mimics the development of limited-growth teeth. 5'-Bromo-2'-deoxyuridine (BrdU) labeling and cytokeratin (CK) 14 and Notch2 immunostaining suggested that the inhibition of inner enamel epithelium growth and the more-active proliferation of the outer enamel epithelium and/or stellate reticulum result in Hertwig's epithelial root sheath formation. Second, we examined the effects of Fgf10 overexpression in the transitional stage of molar germs, which led to the formation of apical bud involving in the inhibition of HERS formation. Taken together, these results suggest that the disappearance of Fgf10 signaling leads to the transition from crown to root formation, owing to the loss of a dental epithelial stem cell compartment.

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  • A non-canonical photopigment, melanopsin, is expressed in the differentiating ganglion, horizontal, and bipolar cells of the chicken retina 査読

    S Tomonari, A Takagi, S Akamatsu, S Noji, H Ohuchi

    DEVELOPMENTAL DYNAMICS234 ( 3 ) 783 - 790   2005年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Vertebrate melanopsin is a photopigment in the eye, required for photoentrainment. Melanopsin is more closely related to opsin proteins found in invertebrates, than to the other photo-pigments. Although the invertebrate melanopsin-like protein is localized in rhabdomeric photoreceptors in the invertebrate eye, it has been shown to be expressed in a subset of retinal ganglion cells in the mouse and in horizontal cells in the frog, indicating its diversified expression pattern in vertebrates. Here we show that two types of melanopsin transcripts are expressed in the developing chicken retina. Melanopsin is firstly expressed by a small subset of ganglion cells, and then prominently expressed by horizontal cells and later by bipolar cells in the developing chicken retina. This suggests that a subset of ganglion, horizontal, and bipolar cells in the chicken retina may have rhabdomeric properties in their origins.

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  • A dual role of FGF10 in proliferation and coordinated migration of epithelial leading edge cells during mouse eyelid development 査読

    H Tao, M Shimizu, R Kusumoto, K Ono, S Noji, H Ohuchi

    DEVELOPMENT132 ( 14 ) 3217 - 3230   2005年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    The development of the eyelid requires coordinated cellular processes of proliferation, cell shape changes, migration and cell death. Mutant mice deficient in the fibroblast growth factor 10 (Fgf10) gene exhibit open-eyelids at birth. To elucidate the roles of FGF10 during eyelid formation, we examined the expression pattern of Fgf10 during eyelid formation and the phenotype of Fgf10-null eyelids in detail. Fgf10 is expressed by mesenchymal cells just beneath the protruding epidermal cells of the nascent eyelid. However, Fgf10-null epithelial cells running though the eyelid groove do not exhibit typical cuboid shape or sufficient proliferation. Furthermore, peridermal clumps are not maintained on the eyelid leading edge, and epithelial extension does not occur. At the cellular level, the accumulation of actin fibers is not observed in the mutant epithelial leading edge. The expression of activin/inhibin beta B (Act beta B/Inhbb) and transforming growth factor a (Tgfa), previously reported to be crucial for eyelid development, is down-regulated in the mutant leading edge, while the onset of sonic hedgehog (Shh) expression is delayed on the mutant eyelid margin. Explant cultures of mouse eyelid primordia shows that the open-eyelid phenotype of the mutant is reduced by exogenous FGF10 protein, and that the expression of Act beta B and Tgfa is ectopically induced in the thickened eyelid epithelium by the FGF10 protein. These results indicate a dual role of FGF10 in mouse eyelid development, for both proliferation and coordinated migration of eyelid epithelial cells by reorganization of the cytoskeleton, through the regulation of activin, TGF alpha and SHH signaling.

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  • Identification of cis-element regulating expression of the mouse Fgf10 gene during inner ear development 査読

    H Ohuchi, A Yasue, K Ono, S Sasaoka, S Tomonari, A Takagi, M Itakura, K Moriyama, S Noji, T Nohno

    DEVELOPMENTAL DYNAMICS233 ( 1 ) 177 - 187   2005年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Fibroblast growth factor (FGF) signaling is crucial for the induction and growth of the ear, a sensory organ that involves intimate tissue interactions. Here, we report the abnormality of Fgf10 null ear and the identification of a cis-regulatory element directing otic expression of Fgf10. In Fgf10 null inner ears, we found that the initial development of semicircular, vestibular, and cochlear divisions is roughly normal, after which there are abnormalities of semicircular canal/cristae and vestibular development. The mutant semicircular disks remain without canal formation by the perinatal stage. To elucidate regulation of the Fgf10 expression during inner ear development, we isolated a 6.6-kb fragment of its 5'-upstream region and examined its transcriptional activity with transgenic mice, using a lacZ-reporter system. From comparison of the mouse sequences of the 6.6-kb fragment with corresponding sequences of the human and chicken Fgf10, we identified a 0.4-kb enhancer sequence that drives Fgf10 expression in the developing inner ear. The enhancer sequences have motifs for many homeodomain-containing proteins (e.g., Prx, Hox, Nkx), in addition to POU-domain factors (e.g., Brn3), zinc-finger transcription factors (e.g., GATA-binding factors), TCF/LEF-1, and a SALAD-interacting protein. Thus, FGF10 signaling is dispensable for specification of otic compartment identity but is required for hollowing the semicircular disk. Furthermore, the analysis of a putative inner ear enhancer of Fgf10 has disclosed a complicated regulation of Fgf10 during inner ear development by numerous transcription factors and signaling pathways. (c) 2005 Wiley-Liss, Inc.

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  • Non-canonical functions of hunchback in segment patterning of the intermediate germ cricket Gryllus bimaculatus 査読

    Taro Mito, Isao Sarashina, Hongjie Zhang, Akihiro Iwahashi, Haruko Okamoto, Katsuyuki Miyawaki, Yohei Shinmyo, Hideyo Ohuchi, Sumihare Noji

    Development132 ( 9 ) 2069 - 2079   2005年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In short and intermediate germ insects, only the anterior segments are specified during the blastoderm stage, leaving the posterior segments to be specified later, during embryogenesis, which differs from the segmentation process in Drosophila, long germ insect. To elucidate the segmentation mechanisms of short and intermediate germ insects, we have investigated the orthologs of the Drosophila segmentation genes in a phylogenetically basal, intermediate germ insect, Gryllus bimaculatus (Gb). Here, we have focused on its hunchback ortholog (Gb'hb), because Drosophila hb functions as a gap gene during anterior segmentation, referred as a canonical function. Gb'hb is expressed in a gap pattern during the early stages of embryogenesis, and later in the posterior growth zone. By means of embryonic and parental RNA interference for Gb'hb, we found the following: (1) Gb'hb regulates Hox gene expression to specify regional identity in the anterior region, as observed in Drosophila and Oncopeltus
    (2) Gb'hb controls germband morphogenesis and segmentation of the anterior region, probably through the pair-rule gene, even-skipped at least
    (3) Gb'hb may act as a gap gene in a limited region between the posterior of the prothoracic segment and the anterior of the mesothoracic segment
    and (4) Gb'hb is involved in the formation of at least seven abdominal segments, probably through its expression in the posterior growth zone, which is not conserved in Drosophila. These findings suggest that Gb'hb functions in a non-canonical manner in segment patterning. A comparison of our results with the results for other derived species revealed that the canonical hb function may have evolved from the non-canonical hb functions during evolution.

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  • FGF19-FGFR4 signaling elaborates lens induction with the FGF8-L-Maf cascade in the chick embryo 査読

    H Kurose, M Okamoto, M Shimizu, T Bito, C Marcelle, S Noji, H Ohuchi

    DEVELOPMENT GROWTH & DIFFERENTIATION47 ( 4 ) 213 - 223   2005年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING ASIA  

    The fibroblast growth factor (FGF) family is known to be involved in vertebrate eye development. However, distinct roles of individual FGF members during eye development remain largely elusive. Here, we show a detailed expression pattern of Fgf19 in chick lens development. Fgf19 expression initiated in the forebrain, and then became restricted to the distal portion of the optic vesicle abutting the future lens placode, where FGF receptor 4 (Fgfr4), a receptor for FGF19, was expressed. Fgf8, a positive regulator for L-Maf, was expressed in a portion of the optic vesicle. To examine the role of FGF19 signaling during early eye development, Fgf19 was misexpressed near the presumptive lens ectoderm; however, no alteration in the expression of lens marker genes was observed. Conversely, a secreted form of FGFR4 was misexpressed to inhibit an FGF19 signal, resulting in the induction of L-Maf expression. To further define the relationship between L-Maf and Fgf19, L-Maf misexpression was performed, resulting in ectopic induction of Fgf19 expression by Hamburger and Hamilton's stage 12/13. Furthermore, misexpression of Fgf8 induced Fgf19 expression in addition to L-Maf. These results suggest that FGF19-FGFR4 signaling plays a role in early lens development in collaboration with FGF8 signaling and L-Maf transcriptional system.

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  • Expression patterns of the homeotic genes Scr, Antp, Ubx, and abd-A during embryogenesis of the cricket Gryllus bimaculatus 査読

    Hongjie Zhang, Yohei Shinmyo, Taro Mito, Katsuyuki Miyawaki, Isao Sarashina, Hideyo Ohuchi, Sumihare Noji

    Gene Expression Patterns5 ( 4 ) 491 - 502   2005年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We have studied embryogenesis of the two-spotted cricket Gryllus bimaculatus as an example of a hemimetabolous, intermediate germ insect, which is a phylogenetically basal insect and may retain primitive features. We observed expression patterns of the orthologs of the Drosophila homeotic genes, Sex combs reduced (Scr), Antennapedia (Antp), Ultrabithorax (Ubx) and abdominal-A (abd-A) during embryogenesis and compared the expression patterns of these genes with the more basal thysanuran insect, Thermobia domestica (the firebrat), and the derived higher dipteran insect, Drosophila melanogaster. Although Scr is expressed commonly in the presumptive posterior maxillary and labial segment in all three insects, the thoracic expression domains vary. Antp is expressed similarly in the three thoracic segments, the limbs, and the anterior abdominal region among these three insects. The early Antp expression in the firebrat and cricket obeys a segmental register in all three thoracic segments, while in Drosophila its initial expression appears in parasegments 4 and 6. Ubx is expressed in the metathoracic (T3) and abdominal segments similarly in the three insects, whereas the expression pattern in the T3 leg differs among them. abd-A is expressed in the posterior compartment of the first abdominal segment and the remaining abdominal segments in all three insects, although its posterior border varies among them. © 2004 Elsevier B.V. All rights reserved.

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  • Location of micropyles and early embryonic development of the two-spotted cricket Gryllus bimaculatus (Insecta, Orthoptera) 査読

    Isao Sarashina, Taro Mito, Michiko Saito, Hiroyuki Uneme, Katsuyuki Miyawaki, Yohei Shinmyo, Hideyo Ohuchi, Sumihare Noji

    Development Growth and Differentiation47 ( 2 ) 99 - 108   2005年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Early embryogenesis of the two-spotted cricket Gryllus bimaculatus was examined by scanning electron microscopy and several fluorescence staining methods, with special reference to these four issues: (i) the location of micropyles
    (ii) the transfer of the female pronucleus following meiosis
    (iii) the timing of cellularization
    and (iv) the process of the germ primordium formation. Between two and four micropyles lie in the mid-ventral region of the egg. The egg nucleus is at the mid-dorsal periphery of the new laid egg, and meiosis resumes and is completed there. The female pronucleus moves to the mid-ventral side, and fertilization occurs there. Energid starts to proliferate and migrates to the periphery of the egg, initiating blastoderm formation. Actin caps surround each superficial nucleus. Cellularization occurs during the blastoderm stage. At a late blastoderm stage, nuclei aggregate in both the posterolateral patch-like regions of the egg to form a germ primordium. The germ primordium looks like a pair of dumbbells. Both the patches shift towards the ventral side and fuse into a germ primordium. The germ primordium contracts to produce a clearly delineated germ band. Observations on distribution patterns of F-actin indicate that, all through the process, the germ primordium retains that unity, and is not separated into two parts.

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  • caudal is required for gnathal and thoracic patterning and for posterior elongation in the intermediate-germband cricket Gryllus bimaculatus 査読

    Yohei Shinmyo, Taro Mito, Takashi Matsushita, Isao Sarashina, Katsuyuki Miyawaki, Hideyo Ohuchi, Sumihare Noji

    Mechanisms of Development122 ( 2 ) 231 - 239   2005年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Although the molecular mechanisms directing anteroposterior patterning of the Drosophila embryo (long-germband mode) are well understood, how these mechanisms were evolved from an ancestral mode of insect embryogenesis remains largely unknown. In order to gain insight into mechanisms of evolution in insect embryogenesis, we have examined the expression and function of the orthologue of Drosophila caudal (cad) in the intermediate-germband cricket Gryllus bimaculatus. We observed that a posterior (high) to anterior (low) gradient in the levels of Gryllus bimaculatus cad (Gb′ cad) transcript was formed in the early-stage embryo, and then Gb′ cad was expressed in the posterior growth zone until the posterior segmentation was completed. Reduction of Gb′ cad expression level by RNA interference resulted in deletion of the gnathum, thorax, and abdomen in embryos, remaining only anterior head. We found that the gnathal and thoracic segments are formed by Gb′ cad probably through the transcriptional regulation of gap genes including Gb′ hunchback and Gb′ Krüppel. Furthermore, Gb′ cad was found to be involved in the posterior elongation, acting as a downstream gene in the Wingless/Armadillo signalling pathways. These findings indicate that Gb′ cad does not function as it does in Drosophila, suggesting that regulatory and functional changes of cad occurred during insect evolution. Since Wnt/Cdx pathways are involved in the posterior patterning of vertebrates, such mechanisms may be conserved in animals that undergo sequential segmentation from the posterior growth zone. © 2004 Elsevier Ireland Ltd. All rights reserved.

    DOI: 10.1016/j.mod.2004.10.001

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  • Mutations in the gene encoding fibroblast growth factor 10 are associated with aplasia of lacrimal and salivary glands 査読

    M Entesarian, H Matsson, J Klar, B Bergendal, L Olson, R Arakaki, Y Hayashi, H Ohuchi, B Falahat, AI Bolstad, R Jonsson, M Wahren-Herlenius, N Dahl

    NATURE GENETICS37 ( 2 ) 125 - 127   2005年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Autosomal dominant aplasia of lacrimal and salivary glands (ALSG; OMIM 180920 and OMIM 103420) is a rare condition characterized by irritable eyes and dryness of the mouth. We mapped ALSG to 5p13. 2- 5q13. 1, which coincides with the gene fibroblast growth factor 10 (FGF10). In two extended pedigrees, we identified heterozygous mutations in FGF10 in all individuals with ALSG. Fgf10(+/-) mice have a phenotype similar to ALSG, providing a model for this disorder. We suggest that haploinsufficiency for FGF10 during a crucial stage of development results in ALSG.

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  • Expression of Fibroblast growth factor 19 (Fgf19) during chicken embryogenesis and eye development, compared with Fgf15 expression in the mouse 査読

    H Kurose, T Bito, T Adachi, M Shimizu, S Noji, H Ohuchi

    GENE EXPRESSION PATTERNS4 ( 6 ) 687 - 693   2004年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The normal development of eyes relies on proper signaling through Fibroblast growth factor (FGF) receptors, but the source and identity of cognate ligands have remained largely unknown. We have found that Fgf19 is expressed in the developing chicken retina. In situ hybridization discloses dynamic expression patterns for Fgf19 in the optic vesicle, lens primordia and retinal horizontal cells. Overall expression pattern of Fgf19 during chicken embryogenesis was also examined: Fgf19 is expressed in the regions associated with cranial placodes induction, boundary regions of rhombomeres, somites, specific groups of neural cells in midbrain, hindbrain, and those derived from epibranchial placodes, and the apical ectodermal ridge of limb buds. Expression pattern of the Fgf19-orthologous gene Fgf15 was further examined in the mouse developing eye. Fgf15 is expressed in the optic vesicle, a subset of progenitor cells of neural retina, and emerging ganglion and amacrine cells during retinogenesis. (C) 2004 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.modgep.2004.04.005

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  • Expression patterns of dachshund during head development of Gryllus bimaculatus (cricket). 査読

    Inoue Y, Miyawaki K, Terasawa T, Matsushima K, Shinmyo Y, Niwa N, Mito T, Ohuchi H, Noji S

    Gene expression patterns : GEP4 ( 6 ) 725 - 731   2004年10月

  • piggyBac-mediated somatic transformation of the two-spotted cricket, Gryllus bimaculatus 査読

    Yohei Shinmyo, Taro Mito, Takashi Matsushita, Isao Sarashina, Katsuyuki Miyawaki, Hideyo Ohuchi, Sumihare Noji

    Development Growth and Differentiation46 ( 4 ) 343 - 349   2004年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Transgenic insects have been artificially produced to study functions of interesting developmental genes, using insect transposons such as piggyBac. In the case of the cricket, however, transgenic animals have not yet been successfully artificially produced. In the present study, we examined whether the piggyBac transposon functions as a tool for gene delivery in embryos of Gryllus bimaculatus. We used either a piggyBac helper plasmid or a helper RNA synthesized in vitro as a transposase source. An excision assay revealed that the helper RNA was more effective in early Gryllus eggs to transpose a marker gene of eGFP than the helper plasmid containing the piggyBac transposase gene driven by the G. bimaculatus actin3/4 promoter. Further, only when the helper RNA was used, somatic transformation of the embryo with the eGFP gene was observed. These results suggest that the piggyBac system with the helper RNA may be effective for making transgenic crickets.

    DOI: 10.1111/j.1440-169x.2004.00751.x

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  • Involvement of Wingless/Armadillo signaling in the posterior sequential segmentation in the cricket, Gryllus bimaculatus (Orthoptera), as revealed by RNAi analysis 査読

    Katsuyuki Miyawaki, Taro Mito, Isao Sarashina, Hongjie Zhang, Yohei Shinmyo, Hideyo Ohuchi, Sumihare Noji

    Mechanisms of Development121 ( 2 ) 119 - 130   2004年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In insects, there are two different modes of segmentation. In the higher dipteran insects (like Drosophila), their segmentation takes place almost simultaneously in the syncytial blastoderm. By contrast, in the orthopteran insects (like Schistocerca (grasshopper)), the anterior segments form almost simultaneously in the cellular blastoderm and then the remaining posterior part elongates to form segments sequentially from the posterior proliferative zone. Although most of their orthologues of the Drosophila segmentation genes may be involved in their segmentation, little is known about their roles. We have investigated segmentation processes of Gryllus bimaculatus, focusing on its orthologues of the Drosophila segment-polarity genes, G. bimaculatus wingless (Gbwg), armadillo (Gbarm) and hedgehog (Gbhh). Gbhh and Gbwg were observed to be expressed in the each anterior segment and the posterior proliferative zone. In order to know their roles, we used RNA interference (RNAi). We could not observed any significant effects of RNAi for Gbwg and Gbhh on segmentation, probably due to functional replacement by another member of the corresponding gene families. Embryos obtained by RNAi for Gbarm exhibited abnormal anterior segments and lack of the abdomen. Our results suggest that GbWg/GbArm signaling is involved in the posterior sequential segmentation in the G. bimaculatus embryos, while Gbwg, Gbarm and Gbhh are likely to act as the segment-polarity genes in the anterior segmentation similarly as in Drosophila. © 2004 Elsevier Ireland Ltd. All rights reserved.

    DOI: 10.1016/j.mod.2004.01.002

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  • Hypotonic buffer induces meiosis and formation of anucleate cytoplasmic islands in the egg of the two-spotted cricket Gryllus bimaculatus 査読

    Isao Sarashina, Yohei Shinmyo, Ayumi Hirose, Katsuyuki Miyawaki, Taro Mito, Hideyo Ohuchi, Tetsuya Horio, Sumihare Noji

    Development Growth and Differentiation45 ( 2 ) 103 - 112   2003年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In insects, egg activation is known to occur in vivo and independently of fertilization, but its mechanisms are poorly understood. To gain understanding of these mechanisms, an attempt was made to activate the egg of Gryllus bimaculatus in vitro. It was found that meiosis resumed and was completed in unfertilized eggs treated with hypotonic buffer. Early developmental processes in activated, unfertilized eggs were investigated and compared with those in fertilized eggs. Mitosis did not progress, resulting in formation of anucleate cytoplasmic islands (pseudoenergids). Development in the activated, unfertilized eggs stopped at this stage and both yolk subdivision and cellularization did not occur. To elucidate the role of the nucleus in the developmental process to the syncytial stage in fertilized eggs, eggs were treated with aphidicolin to inhibit DNA polymerization. It was found that pseudoenergids also formed in these aphidicolin-treated fertilized eggs. These results demonstrate that pseudoenergids can increase in number independently of nuclei, suggesting that the cytoplasm rather than the nucleus plays the primary role in development to the syncytial stage in G. bimaculatus.

    DOI: 10.1034/j.1600-0854.2004.00679.x

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  • Fibroblast growth factor 10 is required for proper development of the mouse whiskers 査読

    H Ohuchi, H Tao, K Ohata, N Itoh, S Kato, S Noji, K Ono

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS302 ( 3 ) 562 - 567   2003年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Fibroblast Growth Factor (FGF) signaling is known to play an important role during cutaneous development. To elucidate the role of FGF10 during whisker formation, we examined the expression of Fgf10 in normal developing whiskers and phenotypes of Fgf10-deficient whiskers. Fgf10 is first expressed in the maxillary process, lateral and medial nasal processes, then in the mesenchymal cells underneath the future whisker placodes, and in the surrounding mesenchyme of developing whiskers. Fgf10-null whiskers exhibit a significant decrease in number and their structure is disorganized as revealed by scanning electron microscopy. Hair follicle marker genes such as Sonic hedgehog, Patched, and Patched 2 are aberrantly expressed in the mutant whiskers. Thus, FGF10 is required for proper whisker development mediated by SHH signaling in the mouse. (C) 2003 Elsevier Science (USA). All rights reserved.

    DOI: 10.1016/S0006-291X(03)00183-9

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  • Extrachromosomal transposition of the transposable element Minos in embryos of the cricket Gryllus bimaculatus. 査読

    Zhang H, Shinmyo Y, Hirose A, Mito T, Inoue Y, Ohuchi H, Loukeris TG, Eggleston P, Noji S

    Development, growth & differentiation44 ( 5 ) 409 - 417   2002年10月

  • Involvement of hedgehog, wingless, and dpp in the initiation of proximodistal axis formation during the regeneration of insect legs, a verification of the modified boundary model. 査読

    Mito T, Inoue Y, Kimura S, Miyawaki K, Niwa N, Shinmyo Y, Ohuchi H, Noji S

    Mechanisms of development114 ( 1-2 ) 27 - 35   2002年6月

  • Correlation of expression patterns of homothorax, dachshund, and Distal-less with the proximodistal segmentation of the cricket leg bud. 査読

    Inoue Y, Mito T, Miyawaki K, Matsushima K, Shinmyo Y, Heanue TA, Mardon G, Ohuchi H, Noji S

    Mechanisms of development113 ( 2 ) 141 - 148   2002年5月

  • Expression patterns of aristaless in developing appendages of Gryllus bimaculatus (cricket). 査読

    Miyawaki K, Inoue Y, Mito T, Fujimoto T, Matsushima K, Shinmyo Y, Ohuchi H, Noji S

    Mechanisms of development113 ( 2 ) 181 - 184   2002年5月

  • FGF10 maintains stem cell compartment in developing mouse incisors. 査読

    Harada H, Toyono T, Toyoshima K, Yamasaki M, Itoh N, Kato S, Sekine K, Ohuchi H

    Development (Cambridge, England)129   1533 - 1541   2002年3月

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  • FGF10 maintains stem cell population during mouse incisor development 査読

    Hidemitsu Harada, Takashi Toyono, Kuniaki Toyoshima, Hideyo Ohuchi

    Connective Tissue Research43 ( 2-3 ) 201 - 204   2002年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Mouse incisors have a cervical loop that gives rise to dental epithelium in the apical region of the tooth germ, in contrast to molars. In a study of formation of the stem-cell compartment, we focused on expression patterns of fibroblast growth factor (Fgf) 10 and Fgf3 in developing mice incisors. At E14, Fgf10 and Fgf3 were coexpressed in the dental papilla. After E16 mesenchymal cells underlying cervical loop expressed Fgf10 but not Fgf3. To illustrate the role of FGF10, we analyzed incisor development of Fgf10-deficient mice. The germs of FGF10-null mice proceeded to cap stage normally. However, at a later stage, the cervical loop was not formed. Functional disorder of FGF10 by a neutralizing anti-FGF10 antibody induced apoptosis in the cervical loop of incisor explants. Recombinant FGF10 rescued the cervical loop from apoptosis. These results show that FGF10 maintains the stem-cell compartment in the developing incisor tooth germ.

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  • Identification of cis-elements regulating expression of Fgf10 during limb development 査読

    H Sasaki, T Yamaoka, H Ohuchi, A Yasue, T Nohno, H Kawano, S Kato, M Itakura, M Nagayama, S Noji

    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY46 ( 7 ) 963 - 967   2002年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:U B C PRESS  

    Fibroblast growth factor 10 (FGF10) is known to be expressed in limb mesenchymal cells and to function as a mesenchymal signaling factor involved in epithelial-mesenchymal interactions during limb development. To elucidate regulation of Fgf10 expression, we isolated the promoter region of Fgf10 containing its 2.0 kb upstream 5'-fragment from the initiation codon and its 0.9 kb downstream fragment. Transcriptional activity of the fragment was examined with transgenic mice, using a lacZ-reporter system. Although no significant expression of the reporter gene was observed for the 0.2 kb 5'-fragment, expression was detected in the apical ectodermal ridge of the limb bud and developing cartilage of the limb for the 2.0 kb and 0.7 kb 5'-fragments, respectively. From comparison of the mouse sequences of the 2.0 kb fragment with corresponding sequences of human and chicken Fgf10, we identified 17 conserved putative enhancer motifs for AER expression and other unidentified expressions. For limb cartilage expression, we found putative enhancer sequences conserved among the three species in the 0.7 kb 5'-fragment. In the fragment, three DNA binding motifs were identified in the mouse and human sequence, although they are not conserved in the corresponding chicken sequence.

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  • A missense mutant myostatin causes hyperplasia without hypertrophy in the mouse muscle

    Masumi Nishi, Akihiro Yasue, Shinichirou Nishimatu, Tsutomu Nohno, Takashi Yamaoka, Mitsuo Itakura, Keiji Moriyama, Hideyo Ohuchi, Sumihare Noji

    Biochemical and Biophysical Research Communications293 ( 1 ) 247 - 251   2002年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Myostatin, which is a member of the TGF-β superfamily, is a negative regulator of skeletal muscle formation. Double-muscled Piedmontese cattle have a C313Y mutation in myostatin and show increased skeletal muscle mass which resulted from an increase of myofiber number (hyperplasia) without that of myofiber size (hypertrophy). To examine whether this mutation in myostatin gene affects muscle development in a dominant negative manner, we generated transgenic mice overexpressing the mutated gene. The transgenic mice exhibited dramatic increases in the skeletal muscle mass resulting from hyperplasia without hypertrophy. In contrast, it has been reported that a myostatin mutated at its cleavage site produces hypertrophy without hyperplasia in the muscle. Thus, these results suggest that (1) the myostatin containing the missense mutation exhibits a dominant negative activity and that (2) there are two types in the dominant negative form of myostatin, causing either hypertrophy or hyperplasia. © 2002 Elsevier Science (USA). All rights reserved.

    DOI: 10.1016/S0006-291X(02)00209-7

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  • Expression patterns of hedgehog, wingless, and decapentaplegic during gut formation of Gryllus bimaculatus (cricket)

    Yoshiko Inoue, Nao Niwa, Taro Mito, Hideyo Ohuchi, Hidefumi Yoshioka, Sumihare Noji

    Mechanisms of Development110   245 - 248   2001年12月

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    We observed expression patterns of hedgehog (hh), wingless (wg), and decapentaplegic (dpp) during gut development of Gryllus bimaculatus (the cricket), a typical hemimetabolous insect, and compared with those observed in Drosophila, a typical holometabolous insect. Gryllus hh(Gbhh) and Gbwg are expressed in both foregut and hindgut, while Gbdpp is expressed only in the hindgut: at the boundaries between the small and large intestine, and between the large intestine and rectum. Although the expression patterns of Gbhh and Gbwg are essentially comparable to those observed in Drosophila, the expression pattern of Gbdpp differs from those of the Drosophila dpp. © 2002 Elsevier Science Ireland Ltd. All rights reserved.

    DOI: 10.1016/S0925-4773(01)00584-6

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  • Differential expression of two BMP antagonists, gremlin and Follistatin, during development of the chick feather bud

    Akihiro Ohyama, Akihiro Ohyama, Fumi Saito, Hideyo Ohuchi, Sumihare Noji

    Mechanisms of Development100   331 - 333   2001年2月

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    Expression of four BMP antagonist genes, noggin, chordin, gremlin and Follistatin, was examined during chick feather development. Although expression of noggin and chordin was not detected, gremlin and Follistatin were expressed differentially in feather buds. The differential expression patterns of gremlin and Follistatin change dynamically from the nascent inter-feather bud region to the posterior domain of the feather bud. Copyright © 2001 Elsevier Science Ireland Ltd.

    DOI: 10.1016/S0925-4773(00)00525-6

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  • Cloning and expression of the chick p63 gene

    Akihiro Yasue, Hirotaka Tao, Tsutomu Nohno, Keiji Moriyama, Sumihare Noji, Hideyo Ohuchi

    Mechanisms of Development100 ( 1 ) 105 - 108   2001年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We have isolated a chick cDNA for p63, a member of the p53 transcription factor family. This cDNA encodes a protein of 582 amino acids for an α isoform in the C-terminal region, while lacking the N-terminal transactivation domain. The chick p63 gene is first expressed in the prospective cutaneous ectoderm at stage 6 and later in the developing epithelia. The p63 expression is intense in specialized epithelial structures, such as apical ectodermal ridge of the limb bud, epithelia of branchial arches and feather buds. Furthermore, we have found that the transcripts are detected in the interdigital epithelium, intersomite epithelium, and epaxial dermamyotome. (C) 2001 Elsevier Science Ireland Ltd.

    DOI: 10.1016/S0925-4773(00)00504-9

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  • FGF10 acts as a major ligand for FGF receptor 2 IIIb in mouse multi-organ development 査読

    H Ohuchi, Y Hori, M Yamasaki, H Harada, K Sekine, S Kato, N Itoh

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS277 ( 3 ) 643 - 649   2000年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC  

    FGF receptor 2 isoform IIIb (FGFR2b), originally discovered as a receptor for FGF7, is known to be an important receptor in vertebrate morphogenesis, because FGFR2b null mice exhibit agenesis or dysgenesis of various organs, which undergo budding and branching morphogenesis. Since FGF7 null mice do not exhibit marked defects in organogenesis, it has been considered that other FGF(s) than FGF7 might function as a major ligand for FGFR2b during organogenesis. One of the candidate ligands is FGF10, because FGF10 binds to FGFR2b with high affinity and the formation of the limb and lung is arrested in FGF10 null mice as found in FGFR2b-deficient mice. Previous analyses of FGF10 null mice revealed that FGF10 is required for limb and lung development. To elucidate the role of FGF10 in wide-range organogenesis, we further analyzed the phenotypes of the FGF10 knockout mice. We found diverse phenotypes closely related to those for FGFR2-deficient mice, which includes the absence of thyroid, pituitary, and salivary glands, while minor defects were observed in the formation of teeth, kidneys, hair follicles, and digestive organs. These results suggest that FGF10 acts as a major ligand for FGFR2b in mouse multi-organ development. (C) 2000 Academic Press.

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  • Correlation of diversity of leg morphology in Gryllus bimaculatus (cricket) with divergence in dpp expression pattern during leg development 査読

    N Niwa, Y Inoue, A Nozawa, M Saito, Y Misumi, H Ohuchi, H Yoshioka, S Noji

    DEVELOPMENT127 ( 20 ) 4373 - 4381   2000年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Insects can be grouped into mainly two categories, holometabolous and hemimetabolous, according to the extent of their morphological change during metamorphosis. The three thoracic legs, for example, are known to develop through two overtly different pathways: holometabolous insects make legs through their imaginal discs, while hemimetabolous legs develop from their leg buds. Thus, how the molecular mechanisms of leg development differ from each other is an intriguing question. In the holometabolous long-germ insect, these mechanisms have been extensively studied using Drosophila melanogaster, However, little is known about the mechanism in the hemimetabolous insect. Thus, we studied leg development of the hemimetabolous short-germ insect, Gryllus bimaculatus (cricket), focusing on expression patterns of the three key signaling molecules, hedgehog (hh), wingless (wg) and decapentaplegic (dpp), which are essential during leg development in Drosophila, In Gryllus embryos, expression of hh is restricted in the posterior half of each leg bud, while dpp and wg are expressed in the dorsal and ventral sides of its anteroposterior (A/P) boundary, respectively. Their expression patterns are essentially comparable with those of the three genes in Drosophila leg imaginal discs, suggesting the existence of the common mechanism for leg pattern formation. However, we found that expression pattern of dpp was significantly divergent among Glyllus, Schistocerca (grasshopper) and Drosophila embryos, while expression patterns of hh and wg are conserved. Furthermore, the divergence was found between the pro/mesothoracic and metathoracic Gryllus leg buds, These observations imply that the divergence in the dpp expression pattern may correlate with diversity of leg morphology.

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  • Involvement of fibroblast growth factor (FGF)18-FGF8 signaling in specification of left-right asymmetry and brain and limb development of the chick embryo 査読

    Hideyo Ohuchi, Sachie Kimura, Miki Watamoto, Nobuyuki Itoh

    Mechanisms of Development95 ( 1-2 ) 55 - 66   2000年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    To elucidate roles of fibroblast growth factors (FGF)18 during vertebrate development, we examined expression patterns of Fgf18 in chick embryos and observed effects of FGF18 protein on the Hensen's node, isthmus, and limb buds. Fgf18 is expressed on the right side of the node before the expression of Fgf8 starts. FGF18 protein can induce expression of Fgf8 on the left side of the node, indicating involvement of both FGFs in specification of left-right asymmetry. In the developing brain, Fgf18 is expressed in the isthmus, following the Fgf8 expression. Since Fgf18 is induced ectopically during formation of the second midbrain by FGF8 protein, both FGFs also elaborate midbrain development. In the limb bud, Fgf18 is expressed in the mesenchyme and ectopic application of FGF18 protein inhibits bone growth in the limb. FGF18 is thus likely an endogenous ligand of FGF receptor 3, whose mutation causes bone dysplasia in humans. These results demonstrate that the FGF18-FGF8 signaling is involved in various organizing activities and the signaling hierarchies between FGF18 and FGF8 seem to change during patterning of different structures. Copyright (C) 2000 Elsevier Science Ireland Ltd.

    DOI: 10.1016/S0925-4773(00)00331-2

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  • Asymmetric expression of antivin/lefty1 in the early chick embryo. 査読

    Ishimaru Y, Yoshioka H, Tao H, Thisse B, Thisse C, Christopher WVE, Hamada H, Ohuchi H, Noji S

    Mechanisms of Development90 ( 1 ) 115 - 118   2000年

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  • Mouse Pitx2 deficiency leads to anomalies of the ventral body wall, heart, extra- and periocular mesoderm and right pulmonary isomerism 査読

    Kunio Kitamura, Hirohito Miura, Sachiko Miyagawa-Tomita, Masako Yanazawa, Yuko Katoh-Fukui, Rika Suzuki, Hideyo Ohuchi, Atuko Suehiro, Yoshiko Motegi, Yoko Nakahara, Shunzo Kondo, Minesuke Yokoyama

    Development126 ( 24 ) 5749 - 5758   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Pitx2, a bicoid-related homeobox gene, is involved in Rieger's syndrome and the left-right (L-R) asymmetrical pattern formation in body plan. In order to define the genomic structure and roles of Pitx2, we analyzed the genomic structure and generated Pitx2-deficient mice with the lacZ gene in the homeobox-containing exon of Pitx2. We were able to show that among three isoforms of Pitx2, Pitx2c shows asymmetrical expression whereas Pitx2a, Pitx2b and Pitx2c show symmetrical expression. In Pitx2(-/-) embryos there was an increase in mesodermal cells in the distal end of the left lateral body wall and an amnion continuous with the lateral body wall thickened in its mesodermal layer. These changes resulted in a failure of ventral body wall closure. In lung and heart in which Pitx2 is expressed asymmetrically, right pulmonary isomerism, atrioventricular canals with prominent swelling, and juxtaposition of the atrium were detected. The hearts failed to develop tricuspid and mitral valves and a common atrioventricular valve forms. Further, dysgenesis of the Pitx2(-/-) extraocular muscle and thickening of the mesothelial layer of cornea were observed in the ocular system where Pitx2 is expressed symmetrically, and these resulted in enophthalmos. The present study shows that Pitx2 expressed in various sites participates in morphogenesis through three types of actions: the involvement of asymmetric Pitx2 expression in the entire morphogenetic process of L-R asymmetric organs
    the involvement of asymmetric Pitx2 expression in the regional morphogenesis of asymmetric organs
    and finally the involvement of symmetric Pitx2 expression in the regional morphogenesis of symmetric organs.

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  • FGF10 can induce Fgf8 expression concomitantly with En1 and R-fng expression in chick limb ectoderm, independent of its dorsoventral specification 査読

    H Ohuchi, T Nakagawa, N Itoh, S Noji

    DEVELOPMENT GROWTH & DIFFERENTIATION41 ( 6 ) 665 - 673   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL SCIENCE ASIA  

    The limb bud has a thickened epithelium at the dorsal-ventral boundary, the apical ectodermal ridge (AER), which sustains limb outgrowth and patterning. A secreted molecule fibroblast growth factor (FGF)10 is involved in inducing Fgf8 expression in the prospective AER and mutual interaction between mesenchymal FGF10 and FGF8 in the AER is essential for limb outgrowth. A secreted factor Wnt7a and a homeobox protein Lmx1 are involved in the dorsal patterning of the limb, whereas a homeobox protein Engrailed 1 (En1) is involved in the dorsal-ventral patterning as well as AER formation. Radical fringe (R-fng), a vertebrate homolog of Drosophila fringe was also found to elaborate AER formation in chicks. However, little is known about the molecular interactions between these factors during AER formation. The present study clarified the relationship between FGF10, Wnt7a, Lmx1, R-fng and En1 during limb development using a foil-barrier insertion experiment. It was found that a foil-barrier inserted into the chick prospective wing mesenchyme lateral to the mesonephric duct blocks AER induction. This experiment was expanded by implanting Fgf10-expressing cells lateral to the barrier and examined whether FGF10 could rescue the expression of the limb-patterning genes reported in AER formation. It was found that FGF10 is sufficient to induce Fgf8 expression in the ectoderm of the foil-inserted limb bud, concomitantly with R-fng and En1 expression. However, FGF10 could not rescue the expression of the dorsal marker genes, Wnt7a or Lmx1. Thus, it is suggested that epithelial factors of En1 and R-fng can induce Fgf8 expression in the limb ectoderm in cooperation with a mesenchymal factor FGF10. Some factor(s) other than FGF10, possibly from the paraxial structures medial to the limb mesoderm, is responsible for the initial dorsal-ventral specification of the limb bud.

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  • FGF7 and FGF10 directly induce the apical ectodermal ridge in chick embryos 査読

    Sayuri Yonei-Tamura, Tetsuya Endo, Hiroshi Yajima, Hideyo Ohuchi, Hiroyuki Ide, Koji Tamura

    Developmental Biology211 ( 1 ) 133 - 143   1999年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Academic Press Inc.  

    During vertebrate limb development, the apical ectodermal ridge (AER) plays a vital role in both limb initiation and distal outgrowth of the limb bud. In the early chick embryo the prelimb bud mesoderm induces the AER in the overlying ectoderm. However, the direct inducer of the AER remains unknown. Here we report that FGF7 and FGF10, members of the fibroblast growth factor family, are the best candidates for the direct inducer of the AER. FGF7 induces an ectopic AER in the flank ectoderm of the chick embryo in a different manner from FGF1, -2, and -4 and activates the expression of Fgf8, an AER marker gene, in a cultured flank ectoderm without the mesoderm. Remarkably, FGF7 and FGF10 applied in the back induced an ectopic AER in the dorsal median ectoderm. Our results suggest that FGF7 and FGF10 directly induce the AER in the ectoderm both of the flank and of the dorsal midline and that these two regions have the competence for AER induction. Formation of the AER of the dorsal median ectoderm in the chick embryo is likely to appear as a vestige of the dorsal fin of the ancestors.

    DOI: 10.1006/dbio.1999.9290

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  • Identification of chick rax/rx genes with overlapping patterns of expression during early eye and brain development 査読

    H Ohuchi, S Tomonari, H Itoh, T Mikawa, S Noji

    MECHANISMS OF DEVELOPMENT85 ( 1-2 ) 193 - 195   1999年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    We have isolated chick rax/rx cDNAs, cRaxL (chick Rax/Rx-like) and cRax, (chick Rax) and examined their expression patterns during early eye and brain development. The cRaxL cDNA encodes a 225 amino acid protein that is most closely related to the zebrafish Rx1 and Rx2. The cRax cDNA encodes a 317 amino acid protein, which shares higher homology with the Xenopus Rx. In addition to the home- odomain, the octapeptide and paired tail domains are conserved between the cRax and other vertebrate Rax/Rx, while cRaxL lacks the octapeptide containing N-terminal region which is conserved among all other members of the rax/rx gene family identified so far. The chick rax/rx genes are expressed in overlapping domains in the anterior neural ectoderm which corresponds to the forebrain and retina field, and later in the optic vesicle. cRax mRNA can be detected earlier than cRaxL prior to the formation of the notochord and its expression domain appears broader than that of cRaxL. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.

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  • FGF-10 is a growth factor for preadipocytes in white adipose tissue 査読

    Masahiro Yamasaki, Hisayo Emoto, Morichika Konishi, Tadahisa Mikami, Hideyo Ohuchi, Kazuwa Nakao, Nobuyuki Itoh

    Biochemical and Biophysical Research Communications258 ( 1 ) 109 - 112   1999年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Academic Press Inc.  

    FGF-10 is a mesenchymal factor affecting epithelial cells during pattern formation. However, the expression and physiological role of FGF-10 in adults remains to be elucidated. We examined the expression of FGF-10 mRNA in a variety of adult rat tissues, and found to be most abundant in white adipose tissue. In white adipose tissue, FGF-10 mRNA was expressed in preadipocytes but not in mature adipocytes. The expression in white adipose tissue during postnatal development was also examined. The expression level was low at postnatal day 10 (P10). However, FGF-10 mRNA was abundantly detected later on (P28 and P48) when white adipose tissue growth was stimulated. We also examined the activity of recombinant FGF-10 for primary rat preadipocytes. FGF-10 showed significant mitogenic activity for primary preadipocytes, but did not affect the differentiation of preadipocytes. The expression profile of FGF-10 mRNA and the activity of FGF-10 reported here indicate that FGF-10, a unique secreted factor produced in white adipose tissue, acts as a growth factor for preadipocytes in white adipose tissues.

    DOI: 10.1006/bbrc.1999.0594

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  • Role of Pax-5 in the regulation of a mid-hindbrain organizer's activity

    Jun ichi Funahashi, Jun ichi Funahashi, Tatsuya Okafuji, Hideyo Ohuchi, Sumihare Noji, Hideaki Tanaka, Harukazu Nakamura, Harukazu Nakamura

    Development Growth and Differentiation41   59 - 72   1999年2月

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    The mes-metencephalic boundary (isthmus) has been suggested to act as an organizer in the development of the optic tectum. Pax-5 was cloned as a candidate for regulator of the organizing center. Isthmus-specific expression of Pax-5 and analogy with the genetic cascade in Drosophila suggest that Pax- 5 may be at a higher hierarchical position in the gene regulation cascade of tectum development. To examine this possibility, a gain-of-function experiment on Pax-5 was carried out. In ovo electroporation on E2 chick brain with the eucaryotic expression vector that encodes chick Pax-5 cDNA was used. Not only was a considerable amount of Pax-5 expressed ectopically in the transfected brain, but irregular bulging of the neuroepithelium was induced in the diencephalon and mesencephalon. At Pax-5 misexpressing sites, uptake of BrdU was increased. Histological examination of E7 transfected brain revealed that Pax-5 caused transdifferentiation of diencephalon into the tectum-like structure. In the bulges of the E7 mesencephalon, differentiation of laminar structure was repressed when compared to the normal side. In transfected embryos, En-2, Wnt-1 and Fgf8 were up-regulated ectopically, and Otx2 was down-regulated in the diencephalon to mesencephalon. Moreover, Ephrin-A2, which is expressed specifically in the tectum with a gradient highest at the caudal end, is suggested to be involved in pathfinding of the retinal fibers, and was induced in the bulges. When the mouse Fgf8 expression vector was electroporated, Pax-5 and chick Fgf8 were also induced ectopically. These results suggest that Pax-5, together with Fgf8, hold a higher position in the genetic hierarchy of the isthmus organizing center and regulate its activity.

    DOI: 10.1046/j.1440-169x.1999.00401.x

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  • Structure and expression of a novel human FGF, FGF-19, expressed in the fetal brain 査読

    Tetsuya Nishimura, Yukari Utsunomiya, Masamitsu Hoshikawa, Hideyo Ohuchi, Nobuyuki Itoh

    Biochimica et Biophysica Acta - Gene Structure and Expression1444 ( 1 ) 148 - 151   1999年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/S0167-4781(98)00255-3

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  • Fgf10 is essential for limb and lung formation 査読

    Keisuke Sekine, Hideyo Ohuchi, Masanori Fujiwara, Masahiro Yamasaki, Tatsuya Yoshizawa, Takashi Sato, Naoko Yagishita, Daisuke Matsui, Yoshihiko Koga, Nobuyuki Itoh, Shigeaki Kato

    Nature Genetics21 ( 1 ) 138 - 141   1999年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The interactions between fibroblast growth factors (FGF) and their receptors have important roles in mediating mesenchymal-epithelial cell interactions during embryogenesis. In particular, Fgf10 is predicted to function as a regulator of brain, lung and limb development on the basis of its spatiotemporal expression pattern in the developing embryo. To define the role of Fgf10, we generated Fgf10-deficient mice. Fgf10(-/-) mice died at birth due to the lack of lung development. Trachea was formed, but subsequent pulmonary branching morphogenesis was disrupted. In addition, mutant mice had complete truncation of the fore- and hindlimbs. In Fgf10(-/-) embryos, limb bud formation was initiated but outgrowth of the limb buds did not occur
    however, formation of the clavicles was not affected. Analysis of the expression of marker genes in the mutant limb buds indicated that the apical ectodermal ridge (AER) and the zone of polarizing activity (ZPA) did not form. Thus, we show here that Fgf10 serves as an essential regulator of lung and limb formation.

    DOI: 10.1038/5096

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  • FGFR2 signaling in normal and limbless chick limb buds 査読

    Gail Lizarraga, Deborah Ferrari, Michael Kalinowski, Hideyo Ohuchi, Sumihare Noji, Robert A. Kosher, Caroline N. Dealy

    Developmental Genetics25 ( 4 ) 331 - 338   1999年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Wiley-Liss Inc.  

    FGF 10 and FGF8, which are reciprocally expressed by the mesoderm and AER of the developing limb bud, have been implicated in limb initiation, outgrowth, and patterning. FGF 10 and FGF8 signal through the FGFR2b and FGFR2c alternative splice isoforms, respectively [Ornitz DM, et al. 1996. J Biol Chem 271:15292-15297
    Igarashi M, et al. 1998. J Biol Chem 273:13230- 13235]. A paracrine signaling loop model has been proposed whereby FGF 10 expressed by limb mesoderm signals via ectodermally restricted FGFR2b to regulate FGF8 expression by the apical ectoderm
    in turn, FGF8 signals via mesodermally restricted FGFR2c to maintain FGF 10 expression [Ohuchi H, et al. 1997. Development 124:2235-2244
    Xu X, et al. 1998. Development 125:753- 765]. To explore this model, we have examined FGFR2b and FGFR2c mRNA expression, using isoform-specific probes during the early stages of development of the chick limb when limb initiation, AER induction, and outgrowth are occurring. We have found that FGFR2b is expressed by limb ectoderm, including the AER, consistent with paracrine signaling of FGF 10. By contrast, FGFR2c is expressed by both mesoderm and ectoderm, indicating that FGF8 has the potential to function in an autocrine as well as paracrine fashion. Indeed, as the limb grows out in response to the AER, FGFR2c expression attenuates in the mesoderm of the progress zone, but is maintained in the AER itself, arguing against exclusive paracrine signaling of FGF8 during limb outgrowth. We also report that transcripts for FGF 10, FGFR2b, and FGFR2c are expressed normally in the limb buds of limbless mutant embryos, which fail to form an AER and do not express FGF8. Furthermore, we detect no mutations in exons specific for the FGFR2c or FGFR2b isoforms in limbless embryos. Since gene targeting has shown that expression of FGF8 in limb ectoderm depends on FGF 10 [Min H, et al. 1998. Genes Dev 12:3156-3161
    Sekine K, et al. 1999. Nature Genet 21:138-141], these results indicate that the product of the limbless gene is required for FGF 10 to induce expression of FGF8.

    DOI: 10.1002/(SICI)1520-6408(1999)25:4<331::AID-DVG7>3.0.CO;2-U

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  • Pitx2, a bicoid-type homeobox gene, is involved in a lefty-signaling pathway in determination of left-right asymmetry

    Hidefumi Yoshioka, Chikara Meno, Kazuko Koshiba, Minoru Sugihara, Hiroyuki Itoh, Yoshiyasu Ishimaru, Takashi Inoue, Hideyo Ohuchi, Elena V. Semina, Jeffrey C. Murray, Hiroshi Hamada, Sumihare Noji

    Cell94   299 - 305   1998年8月

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    Signaling molecules such as Activin, Sonic hedgehog, Nodal, Lefty, and Vg1 have been found to be involved in determination of left-right (L-R) asymmetry in the chick, mouse, or frog. However, a common signaling pathway has not yet been identified in vertebrates. We report that Pitx2, a bicoid- type homeobox gene expressed asymmetrically in the left lateral plate mesoderm, may be involved in determination of L-R asymmetry in both mouse and chick. Since Pitx2 appears to be downstream of lefty-1 in the mouse pathway, we examined whether mouse Lefty proteins could affect the expression of Pitx2 in the chick. Our results indicate that a common pathway from lefty-1 to Pitx2 likely exists for determination of L-R asymmetry in vertebrates.

    DOI: 10.1016/S0092-8674(00)81473-7

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  • Correlation of wing-leg identity in ectopic FGF-induced chimeric limbs with the differential expression of chick Tbx5 and Tbx4. 査読

    Ohuchi H, Takeuchi J, Yoshioka H, Ishimaru Y, Ogura K, Takahashi N, Ogura T, Noji S

    Development125 ( 1 ) 51 - 60   1998年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Essential roles of the winged helix transcription factor MFH-1 in aortic arch patterning and skeletogenesis 査読

    K Iida, H Koseki, H Kakinuma, N Kato, Y Mizutani-Koseki, H Ohuchi, H Yoshioka, S Noji, K Kawamura, Y Kataoka, F Ueno, M Taniguchi, N Yoshida, T Sugiyama, N Miura

    DEVELOPMENT124 ( 22 ) 4627 - 4638   1997年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Mesenchyme Fork Head-1 (MFH-1) is a forkhead (also called winged helix) transcription factor defined by a common 100-amino acid DNA-binding domain, MFH-1 is expressed in non-notochordal mesoderm in the prospective trunk region and in cephalic neural-crest and cephalic mesoderm-derived mesenchymal cells in the prechordal region of early embryos, Subsequently, strong expression is localized in developing cartilaginous tissues, kidney and dorsal aortas, To investigate the developmental roles of MFH-1 during embryogenesis, mice lacking the MFH-1 locus were generated by targeted mutagenesis, MFH-1-deficient mice died embryonically and perinatally, and exhibited interrupted aortic arch and skeletal defects in the neurocranium and the vertebral column. Interruption of the aortic arch seen in the mutant mice was the same as in human congenital anomalies. These results suggest that MFH-1 has indispensable roles during the extensive remodeling of the aortic arch in neural-crest-derived cells and in skeletogenesis in cells derived from the neural crest and the mesoderm.

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  • Pax-6 is involved in the specification of hindbrain motor neuron subtype

    Noriko Osumi, Noriko Osumi, Arisa Hirota, Hideyo Ohuchi, Masato Nakafuku, Tadahiro Iimura, Shigeru Kuratani, Michio Fujiwara, Sumihare Noji, Kazuhiro Eto

    Development124   2961 - 2972   1997年9月

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    Pax-6 is a member of the vertebrate Pax gene family, which is structurally related to the Drosophila pair-rule gene, paired. In mammals, Pax-6 is expressed in several discrete domains of the developing CNS and has been implicated in neural development, although its precise role remains elusive. We found a novel Small eye rat strain (rSey2) with phenotypes similar to mouse and rat Small eye. Analyses of the Pax-6 gene revealed one base (C) insertion in an exon encoding the region downstream of the paired box of the Pax-6 gene, resulting in generation of truncated protein due to the frame shift. To explore the roles of Pax-6 in neural development, we searched for abnormalities in the nervous system in rSey2 homozygous embryos. rSey2/rSey2, exhibited abnormal development of motor neurons in the hindbrain. The Islet-1-positive motor neurons were generated just ventral to the Pax-6-expressing domain both in the wild-type and mutant embryos. However, two somatic motor (SM) nerves, the abducent and hypoglossal nerves, were missing in homozygous embryos, By retrograde and anterograde labeling, we found no SM-type axonogenesis (ventrally growing) in the mutant postotic hindbrain, though branchiomotor and visceral motor (BM/VM)-type axons (dorsally growing) were observed within the neural tube. To discover whether the identity of these motor neuron subtypes was changed in the mutant, we examined expression of LIM homeobox genes, Islet-1, Islet-2 and Lim-3. At the postotic levels of the hindbrain, SM neurons expressed all the three LIM genes, whereas BM/VM-type neurons were marked by Islet-1 only. In the Pax-6 mutant hindbrain, Islet-2 expression was specifically missing, which resulted in the loss of the cells harboring the postotic hindbrain SM-type LIM code (Islet-1 + Islet-2 + Lim-3). Furthermore, we found that expression of Wnt-7b, which overlapped with Pax-6 in the ventrolateral domain of the neural tube, was also specifically missing in the mutant hindbrain, while it remained intact in the dorsal non-overlapping domain. These results strongly suggest that Pax-6 is involved in the specification of subtypes of hindbrain motor neurons, presumably through the regulation of Islet-2 and Wnt-7b expression.

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  • Fibroblasts expressing Sonic hedgehog induce osteoblast differentiation and ectopic bone formation

    Naoki Kinto, Naoki Kinto, Masahiro Iwamoto, Motomi Enomoto-Iwamoto, Sumihare Noji, Hideyo Ohuchi, Hidefumi Yoshioka, Hiroko Kataoka, Yasuhiro Wada, Yasuhiro Wada, Gao Yuhao, Hideaki E. Takahashi, Shusaku Yoshiki, Akira Yamaguchi

    FEBS Letters404   319 - 323   1997年3月

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    We investigated the role of Sonic hedgehog (SHH) in osteoblast differentiation and bone formation. The numbers of ALP-positive cells in the mouse fibroblastic cell line C3H10T1/2 and the mouse osteoblastic cell line MC3T3-E1 were increased by co-culture with chicken fibroblasts transfected with chicken Shh cDNA encoding amino-terminal peptide (Shh-N). The conditioned medium of Shh-N-RCAS-transfected chicken fibroblast cultures also significantly increased ALP activity in both C3H10T1/2 and MC3T3-E1 cells. Intramuscular transplantation of Shh-N-RCAS-transfected chicken fibroblasts into athymic mice induced ectopic bone formation. These results indicate that SHH induces osteoblast differentiation and ectopic bone formation.

    DOI: 10.1016/S0014-5793(97)00014-8

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  • Corelation between Distal-less expression patterns and structures of appendages in development of the two-soptted cricket, Gryllus bimaculatus

    Nao Niwa, Mariko Saitoh, Hideyo Ohuchi, Hidefumi Yoshioka, Sumihare Noji

    Zool. Sci.14 ( 1 ) 115 - 125   1997年

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  • The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor 査読

    H. Ohuchi, T. Nakagawa, A. Yamamoto, A. Araga, T. Ohata, Y. Ishimaru, H. Yoshioka, T. Kuwana, T. Nohno, M. Yamasaki, N. Itoh, S. Noji

    Development124 ( 11 ) 2235 - 2244   1997年

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    記述言語:英語  

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  • INDUCTION OF AN ADDITIONAL LIMB BY FGF AND DUPLICATION OF DIGITS BY SONIC HEDGEHOG 査読

    H OHUCHI, H YOSHIOKA, S NOJI

    SEIKAGAKU67 ( 10 ) 1232 - 1236   1995年10月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN BIOCHEMICAL SOC  

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  • NEW GENE, NEL, ENCODING A M(R) 93-K PROTEIN WITH EGF-LIKE REPEATS IS STRONGLY EXPRESSED IN NEURAL TISSUES OF EARLY-STAGE CHICK-EMBRYOS

    S MATSUHASHI, S NOJI, E KOYAMA, F MYOKAI, H OHUCHI, S TANIGUCHI, K HORI

    DEVELOPMENTAL DYNAMICS203 ( 2 ) 212 - 222   1995年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    A new gene, nel, was isolated from a 9-day-old chick embryonic cDNA library. The gene encodes a protein of 835 amino acids (93,407 M(r)) consisting of two hydrophobic domains presumed to be the signal and transmembrane sequences, a histidine rich domain, two repeats of a cysteine rich structure similar to the C-terminal domain of von Willebrand factor, five EGF-like repeats, and again two repeats of the cysteine rich sequence similar to the C-terminal domain of von Willebrand factor in the presumed cytoplasmic domain. The expression of the nel gene was studied by Northern blot and in situ hybridization analyses of chick embryos. The mRNA of the gene was found in all tissues of 10- and 17-day-old embryos by Northern blot hybridization. Among the tissues examined, the level in the brain was highest and increased with age. After hatching, gene expression was retained in the brain at about the same level found in old embryos, increased in the retina, and disappeared from the other tissues. In situ hybridization with a nel gene probe revealed that the gene was strongly expressed in neural tissues such as brain, spinal cord, and dorsal root ganglia of early embryos. Gene expression was observed in the mantle layer of the neurepithelium of the brain and of the spinal cord. Gene expression in early embryos was not restricted to the neural tissues, but was also detected in the cells around cartilage, myocardium, lung mesenchymal cells, and in the liver, etc. One band of about 4.5 Kb mRNA was detected in all tissues and stages by Northern blot hybridization analysis. The possible function of the gene is discussed. (C) 1995 Wiley-Liss, Inc.

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  • FGF Can Induce Outgrowth of Somatic Mesoderm both Inside and Outside of Limb-Forming Regions

    Tatsuo Mima, Hideyo Ohuchi, Sumihare Noji, Takashi Mikawa

    Developmental Biology167   617 - 620   1995年2月

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    In the vertebrate embryo, only somatopleural cells in the limb-forming region are released from the mesodermal layer and undergo outgrowth from the embryonic body to form the limb bud. Molecular signals which regulate limb bud induction are unknown to date. In the present study we examined the ability of fibroblast growth factor (FGF) to induce limb bud formation in chicken embryos. A replication-defective retrovirus encoding FGF type 4 with a reporter, bacterial β-galactosidase, was microinjected into lateral plate mesoderm inside and outside limb-forming regions. Effects of the ectopic and precocious expression of FGF were assessed at various stages after infection. Here we report that somatic mesodermal cells in both flank and limb-forming regions can respond to FGF and induce limb bud-like outgrowth. The supernumerary limb bud induced within a limb-forming region differentiated into extralimb structures. These results strongly suggest potential roles of FGF signaling for induction of limb bud formation. © 1995 Academic Press. All rights reserved.

    DOI: 10.1006/dbio.1995.1053

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  • Involvement of growth factors and their receptors in morphogenesis of the chick limb

    T. Nohno, H. Ohuchi, H. Yoshioka, S. Noji

    Gann Monographs on Cancer Research42   79 - 90   1994年1月

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    Vertebrate limb morphogenesis has been investigated as a model system of pattern formation. Recent results suggest that peptide growth factors are involved in the limb morphogenesis. Members of the fibroblast growth factor family stimulate proliferation of the limb mesenchymal cells, while members of the transforming growth factor (TGF)-β family, in contrast, inhibit the proliferation. The antagonistic effect between the two families is probably essential for establishment of both anteroposterior and proximodistal axes. The Wnt and TGF-β families may be related to morphogenesis of the dorsoventral axis. The three families are also involved in chondrogenesis, osteogenesis, and feather formation. In this review, possible roles of such growth factors and their receptors in morphogenesis of the limb are discussed.

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  • Differential expression of three chick FGF receptor genes, FGFR1, FGFR2 and FGFR3, in limb and feather development.

    S. Noji, E. Koyama, F. Myokai, T. Nohno, H. Ohuchi, K. Nishikawa, S. Taniguchi

    Progress in clinical and biological research383 B   645 - 654   1993年12月

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    Expression patterns of three fibroblast growth factor receptor genes, FGFR1 (cek1), FGFR2 (cek3) and FGFR3 (cek2), were observed in limb and feather morphogenesis. Expression of FGFR1 was observed in the mesoderm of limb bud, in the mesenchyme just underneath the feather placode, and then in the anterior mesenchyme of the feather bud. While expression of FGFR2 was observed in both surface ectoderm and mesenchymal aggregates corresponding to the future bones of the limb, the mesenchyme between the feather placode, and surface ectoderm of feather buds. Expression of FGFR3 was observed rather ubiquitously over mesoderm of limb and feather buds. Differential expression of these FGF receptor genes suggested that differential roles of these receptors in epithelia-mesenchymal interactions of limb and feather morphogenesis.

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  • A MUTATION IN THE PAX-6 GENE IN RAT SMALL-EYE IS ASSOCIATED WITH IMPAIRED MIGRATION OF MIDBRAIN CREST CELLS 査読

    T MATSUO, N OSUMIYAMASHITA, S NOJI, H OHUCHI, E KOYAMA, F MYOKAI, N MATSUO, S TANIGUCHI, H DOI, S ISEKI, Y NINOMIYA, M FUJIWARA, T WATANABE, K ETO

    NATURE GENETICS3 ( 4 ) 299 - 304   1993年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING CO  

    The rat small eye strain (rSey) lacks eyes and nose in the homozygote, an is similar the mouse Sey strain with mutations in the Pax-6 gene. We isolated Pax-6 cDNA clones from an rSey homozygote library, and found an internal deletion of about 600 basepairs in the serine/threonine-rich domain. At the genomic level, a single base (G) insertion in an exon generates an abnormal 5' donor splice site, thereby producing the truncated mRNA. Anterior midbrain crest cells in the homozygous rSey embryos reached the eye rudiments but did not migrate any further to the nasal rudiments, suggesting that the Pax-6 gene is involved in conducting migration of neural crest cells from the anterior midbrain.

    DOI: 10.1038/ng0493-299

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    その他リンク: http://orcid.org/0000-0003-0548-6452

  • Expression pattern of fibroblast growth factor receptor 1 gene during chickeye development 査読

    H. Ohuchi

    Journal of Japanese Ophthalmological Society97 ( 3 ) 304 - 309   1993年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • Expression pattern of fibroblast growth factor receptor 2 gene during chick eye development 査読

    H. Ohuchi

    Journal of Japanese Ophthalmological Society97 ( 5 ) 563 - 568   1993年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • EXPRESSION PATTERNS OF THE ACTIVIN RECEPTOR IIA AND IIB GENES DURING CHICK LIMB DEVELOPMENT

    T NOHNO, S NOJI, E KOYAMA, F MYOKAI, H OHUCHI, K NISHIKAWA, S SUMITOMO, S TANIGUCHI, T SAITO

    LIMB DEVELOPMENT AND REGENERATION, PT B383   705 - 714   1993年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:WILEY-LISS, INC  

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▼全件表示

書籍等出版物

  • The Cricket as a Model Organism

    Hideyo Ohuchi, Tetsuya Bando, Taro Mito, Sumihare Noji( 担当: 分担執筆 ,  範囲: Chapter 4: Eye Development and Photoreception of a Hemimetabolous Insect, Gryllus bimaculatus)

    Springer Japan KK  2017年 

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  • 光と生命の事典(編集:真嶋哲朗、飯野盛利、七田芳則、藤堂剛)

    大内淑代( 担当: 分担執筆 ,  範囲: 脳内光受容体)

    朝倉書店  2016年2月  ( ISBN:9784254171617

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    総ページ数:422頁   担当ページ:180頁-181頁   記述言語:日本語 著書種別:事典・辞書

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  • 発生生物学 生物はどのように形づくられるか

    Lewis Wolpert 著, 大内淑代, 野地澄晴( 担当: 共訳)

    丸善出版  2013年7月  ( ISBN:9784621086896

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  • マウス表現型解析 失敗しないステップ・バイ・ステップアプローチ Mouse Phenotypes, A Handbook of Mutation Analysis, by V. E. Papaioannou & R. R. Behringer, CSHL Press

    大内淑代, 井関祥子( 担当: 共訳)

    メデイカルサイエンスインターナショナル  2006年 

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MISC

  • Congenital eye anomalies: More mosaic than thought? 査読

    Ohuchi H, Sato K, Habuta M, Fujita H, Bando T

    Congenital anomalies59 ( 3 ) 56 - 73   2019年5月

  • Molecular mechanisms of limb regeneration: insights from regenerating legs of the cricket Gryllus bimaculatus. 査読 国際誌

    Tetsuya Bando, Taro Mito, Yoshimasa Hamada, Yoshiyasu Ishimaru, Sumihare Noji, Hideyo Ohuchi

    The International journal of developmental biology62 ( 6-7-8 ) 559 - 569   2018年

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    記述言語:英語  

    This review summarizes recent advances in leg regeneration research, focusing on the cricket Gryllus bimaculatus. Recent studies have revealed molecular mechanisms on blastema formation, establishment of positional information, and epigenetic regulation during leg regeneration. Especially, these studies have provided molecular bases in classical conceptual models such as the polar coordinate model, the intercalation model, the boundary model, the steepness model, etc., which were proposed to interpret regeneration processes of the cockroach legs. When a leg is amputated, a blastema is formed through the activation of the Janus-kinase (Jak)/Signal-Transduction-and-Activator-of-Transcription (STAT) pathway. Subsequently, the Hedgehog/Wingless/Decapentaplegic/Epidermal-growth-factor pathways instruct distalization in the blastema, designated as the molecular boundary model. Downstream targets of this pathway are transcription factors Distal-less (Dll) and dachshund (dac), functioning as key regulators of proximodistal pattern formation. Dll and dac specify the distal and proximal regions in the blastema, respectively, through the regulation of tarsal patterning genes. The expression of leg patterning genes during regeneration may be epigenetically controlled by histone H3K27 methylation via Enhancer-of-zeste and Ubiquitously-transcribed-tetratricopeptide-repeat-gene-X-chromosome. For the molecular mechanism of intercalation of the missing structures between the amputated position and the most distal one, Dachsous/Fat (Ds/Ft) steepness model has been proposed, in which the Ds/Ft pathway maintains positional information and determines leg size through dac expression. This model was theoretically verified to interpret the experimental results obtained with cricket legs. Availability of whole-genome sequence information, regeneration-dependent RNA interference, and genome editing technique will have the cricket be an ideal model system to reveal gene functions in leg regeneration.

    DOI: 10.1387/ijdb.180048ho

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  • Preface

    Hadley Wilson Horch, Taro Mito, Aleksandar Popadić, Hideyo Ohuchi, Sumihare Noji

    The Cricket as a Model Organism: Development, Regeneration, and Behavior   v - vi   2017年1月

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    記述言語:英語   出版者・発行元:Springer Japan  

    DOI: 10.1007/978-4-431-56478-2

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  • The cricket as a model organism: Development, regeneration, and behavior

    Hadley Wilson Horch, Taro Mito, Aleksandar Popadić, Hideyo Ohuchi, Sumihare Noji

    The Cricket as a Model Organism: Development, Regeneration, and Behavior   1 - 376   2017年1月

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    記述言語:英語   出版者・発行元:Springer Japan  

    This book covers a broad range of topics about the cricket from its development, regeneration, physiology, nervous system, and behavior with remarkable recent updates by adapting the new, sophisticated molecular techniques including RNAi and other genome editing methods. It also provides detailed protocols on an array of topics and for basic experiments on the cricket. While the cricket has been one of the best models for neuroethological studies over the past 60 years, it has now become the most important system for studying basal hemimetabolous insects. The studies of Gryllus and related species of cricket will yield insight into evolutionary features that are not evident in other insect model systems, which mainly focus on holometabolous insects such as Drosophila, Tribolium, and Bombyx. Research on crickets and grasshoppers will be important for the development of pest-control strategies, given that some of the most notorious pests also belong to the order Orthoptera. At the same time, crickets possess an enormously high “food conversion efficiency”, making them a potentially important food source for an ever-expanding human population. This volume provides a comprehensive source of information as well as potential new applications in pest management and food production of the cricket. It will inspire scientists in various disciplines to use the cricket model system to investigate interesting and innovative questions.

    DOI: 10.1007/978-4-431-56478-2

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  • 組織形成因子 Fgf10 招待

    大内淑代

    岡山医学会雑誌125 ( 3 ) 229 - 234   2013年12月

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(大学・研究所紀要)  

    DOI: 10.4044/joma.125.229

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  • Molecular Aspects of Eye Evolution and Development: From the Origin of Retinal Cells to the Future of Regenerative Medicine 査読

    Hideyo Ohuchi

    ACTA MEDICA OKAYAMA67 ( 4 ) 203 - 212   2013年8月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:OKAYAMA UNIV MED SCHOOL  

    A central issue of evolutionary developmental biology is how the eye is diverged morphologically and functionally. However, the unifying mechanisms or schemes that govern eye diversification remain unsolved. In this review, I first introduce the concept of evolutionary developmental biology of the eye with a focus on photoreception, the fundamental property of retinal cells. Second, I summarize the early development of vertebrate eyes and the role of a homeobox gene, Lhx1, in subdivision of the retina into 2 domains, the neural retina and retinal pigmented epithelium of the optic primordium. The 2 retinal domains are essential components of the eye as they are found in such prototypic eyes as the extant planarian eye. Finally, I propose the presence of novel retinal cell subtypes with photosensory functions based on our recent work on atypical photopigments (opsins) in vertebrates. Since human diseases are attributable to the aberration of various types of cells due to alterations in gene expression, understanding the precise mechanisms of cellular diversification and unraveling the molecular profiles of cellular subtypes are essential to future regenerative medicine.

    DOI: 10.18926/AMO/51064

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  • Wakayama Symposium: Epithelial-Mesenchymal Interactions in Eyelid Development 査読

    Hideyo Ohuchi

    OCULAR SURFACE10 ( 4 ) 212 - 216   2012年10月

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    記述言語:英語   出版者・発行元:ETHIS COMMUNICATINS, INC  

    Various congenital anomalies of the eyelids can result from abnormal tissue proliferation, fusion, and reopening of the eyelids. Therefore, it is important to study the molecular mechanisms underlying eyelid development, focusing on cell behaviors. Mammalian eyelid development occurs in four steps: specification, growth, epithelial fusion, and reopening. Epithelial-mesenchymal interactions are fundamental processes during eyelid formation, and epithelial factors functioning within the eyelid epithelium are also critical. Studies with mutant and genetically modified mice have revealed that various signaling pathways and transcriptional factors are involved in mouse eyelid development. In this review, eyelid morphogenetic factors or pathways are described, as revealed by their mutant phenotype, eye-open at birth (EOB). These include FGFR2b-FGF10, EGFR-ERK, MEKK-JNK, BMP, Shh, Wnt, GPR48, Jun, Forkhead, and Grainyhead.

    DOI: 10.1016/j.jtos.2012.07.005

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  • The advent of RNA interference in Entomology

    Taro Mito, Taro Nakamura, Tetsuya Bando, Hideyo Ohuchi, Sumihare Noji

    ENTOMOLOGICAL SCIENCE14 ( 1 ) 1 - 8   2011年1月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:WILEY-BLACKWELL  

    RNA interference (RNAi) is a cellular process by which an mRNA is targeted for degradation by a small interfering RNA that contains a strand complementary to a fragment of the target mRNA, resulting in sequence specific inhibition of gene expression. The discovery of RNAi enabled the use of loss-of-function analyses in many non-model insects other than Drosophila to elucidate the roles of specific genes. The RNAi approach has been widely used on insects in several fields, including embryogenesis, pattern formation, reproduction, biosynthesis and behavior. The increasing availability of insect genomes has made the RNAi technique an indispensable technique for characterizing gene functions in insects. Here we review the current status of RNAi-based experiments in insects and the applications of RNAi for species-specific insecticides, focusing on non-drosophilid insects. We also identify future applications for RNAi-based studies in Entomology.

    DOI: 10.1111/j.1479-8298.2010.00408.x

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  • Dynamics of cellular aggregation and differentiation during early development of transgenic crickets

    Taro Nakamura, Taro Mito, Masato Yoshizaki, Tetsuya Bando, Kimio Tanaka, Hideyo Ohuchi, Sumihare Noji

    MECHANISMS OF DEVELOPMENT126   S254 - S254   2009年8月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.mod.2009.06.666

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  • Dual splice variants of autotaxin control chicken diencephalon-mesencephalon development downstream of Pax6

    Hideyo Ohuchi, Hitomi Fukui, Akane Matsuyo, Sayuri Tomonari, Sumihare Noji

    MECHANISMS OF DEVELOPMENT126   S74 - S74   2009年8月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.mod.2009.06.077

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  • Regulation of leg size and shape by the Dachsous/Fat signalling pathway during regeneration in Gryllus bimaculatus

    Tetsuya Bando, Taro Mito, Yuko Maeda, Taro Nakamura, Fumiaki Ito, Takahito Watanabe, Hideyo Ohuchi, Sumihare Noji

    MECHANISMS OF DEVELOPMENT126   S293 - S294   2009年8月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.mod.2009.06.797

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  • Involvement of the Hedgehog/Engrailed signaling in regeneration of the posterior structures in the amputated leg of the cricket Gryllus bimaculatus

    Katsuyuki Miyawaki, Taro Mito, Taro Nakamura, Tetsuya Bando, Kimio Tanaka, Hideyo Ohuchi, Sumihare Noji

    MECHANISMS OF DEVELOPMENT126   S294 - S294   2009年8月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.mod.2009.06.799

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  • Molecular and cellular basis of regeneration and tissue repair: Dissecting insect leg regeneration through RNA interference

    T. Nakamura, T. Mito, T. Bando, H. Ohuchi, S. Noji

    Cellular and Molecular Life Sciences65 ( 1 ) 64 - 72   2008年1月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等  

    Nymphs of hemimetabolous insects such as cockroaches and crickets exhibit a remarkable capacity for regenerating complex structures from damaged legs. Until recent years, however, approaches to elucidate the molecular mechanisms underlying the leg regeneration process have been lacking. Taking the cricket Gryllus bimaculatus as a model, we found that a phenotype related to regeneration frequently appears during leg regeneration, even though no phenotype is induced by RNA interference (RNAi) in the cricket nymph, designated as regeneration-dependent RNAi. Since then, we have investigated the functions of various genes encoding signaling factors and cellular adhesion proteins like Fat and Dachsous during leg regeneration. In this review, we summarize the classical knowledge about insect leg regeneration and introduce recent advances concerning the signaling cascades required for regenerating a leg. Our results provide clues to the mechanisms of regeneration which are relevant to vertebrate systems. © 2007 Birkhaueser.

    DOI: 10.1007/s00018-007-7432-0

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  • Warts, Expanded and Dachs are required for proper regeneration of amputated legs in the cricket, Gryllus bimaculatus

    Yuko Maeda, Tetsuya Bando, Taro Nakamura, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    GENES & GENETIC SYSTEMS82 ( 6 ) 532 - 532   2007年12月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:GENETICS SOC JAPAN  

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  • Fat and dachsous protocadherin are required for proper proximodistal and circumferential regeneration of amputated legs in the cricket, Gryllus bimaculatus

    Tetsuya Bando, Yuko Maeda, Taro Nakamura, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    GENES & GENETIC SYSTEMS82 ( 6 ) 532 - 532   2007年12月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:GENETICS SOC JAPAN  

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  • ニワトリ神経発生時のautotaxinの機能的研究(Functional studies of Autotaxin during chicken neural development)

    福井 ひと美, 岡本 麻友美, 山中 瑞恵, 佐伯 智佳子, 尾ノ井 基嘉, 松田 洋尚, 湯藤 嘉文, 田中 将之, 青木 淳賢, 新井 洋由, 野地 澄晴, 大内 淑代

    日本発生生物学会・日本細胞生物学会合同大会要旨集40回・59回   104 - 104   2007年5月

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    記述言語:英語   出版者・発行元:日本発生生物学会・日本細胞生物学会  

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  • Involvement of fragile X mental retardation (fmr) in stridulation of the two-spotted cricket (gryllus bimaculatus)

    Aska Hamada, Eri Honda-Sumi, Taro Mito, Hideyo Ohuchi, Sumihare Noji

    ZOOLOGICAL SCIENCE23 ( 12 ) 1222 - 1222   2006年12月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ZOOLOGICAL SOC JAPAN  

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  • Involvement of EGF and Wnt signalling in regeneration of cricket legs, as revealed by RNAi

    S. Noji, T. Nakamura, Y. Tanaka, T. Mito, H. Ohuchi

    MECHANISMS OF DEVELOPMENT122   S130 - S130   2005年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

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  • Roles of FGF10 in mouse eyelid development

    H. Ohuchi, H. Tao, M. Shimizu, R. Kusumoto, K. Ono, S. Noji

    MECHANISMS OF DEVELOPMENT122   S53 - S53   2005年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

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  • Non-canonical regulatory networks of segmentation genes in the intermediate germ cricket Gryllus bimaculatus

    T. Mito, H. Ohuchi, S. Noji

    MECHANISMS OF DEVELOPMENT122   S157 - S157   2005年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE BV  

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  • A missense mutant myostatin causes hyperplasia without hypertrophy in the mouse muscle (vol 293, pg 247, 2002)

    M Nishi, A Yasue, S Nishimatu, T Nohno, T Yamaoka, M Itakura, K Moriyama, H Ohuchi, S Noji

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS318 ( 3 ) 801 - 801   2004年6月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    DOI: 10.1016/j.bbrc.2004.04.091

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  • A missense mutation of myostatin causes hyperplasia in skeletal muscles.

    M Nishi, A Yasue, H Ohuchi, S Noji, K Moriyama

    JOURNAL OF DENTAL RESEARCH82   403 - 403   2003年12月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:INT AMER ASSOC DENTAL RESEARCHI A D R/A A D R  

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  • The role of myostatin in skeletal muscle formation.

    M. Nishi, A. Yasue, H. Ohuchi, S. Noji, K. Moriyama

    JOURNAL OF DENTAL RESEARCH82   B326 - B326   2003年6月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:INT AMER ASSOC DENTAL RESEARCHI A D R/A A D R  

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  • FGF10 is a mesenchymally derived stimulator for epidermal development in the chick embryonic skin.

    Tao Hirotaka, Yoshimoto Yasuko, Yoshioka Hidefumi, Nohno Tsutomu, Noji Sumihare, Ohuchi Hideyo

    Mechanisms of development116 ( 1-2 ) 39 - 49   2002年8月

  • Roles of FGF18 in chick limb development

    H Ohuchi, F Saitoh, N Itoh, S Noji

    DEVELOPMENTAL DYNAMICS219 ( 3 ) 454 - 454   2000年11月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:WILEY-LISS  

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  • Limb development and regeneration of the cricket leg: Expression patterns of hedgehog, wingless and decapentaplegic

    S Noji, Y Inoue, T Mito, H Ohuchi

    DEVELOPMENTAL DYNAMICS219 ( 3 ) 440 - 441   2000年11月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:WILEY-LISS  

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  • Fibroblast-growth-factor-induced additional limbs in the study of initiation of limb formation, limb identity, myogenesis, and innervation

    Hideyo Ohuchi, Sumihare Noji

    Cell and Tissue Research296 ( 1 ) 45 - 56   1999年

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等  

    In this review, we focus on the additional limb induced by members of the fibroblast growth factor (FGF) family in the flank of chick embryos. The 'additional limb' was first reported 73 years ago by Balinsky in 1925. He grafted otic vesicle to the flank of newt embryos and observed the formation of the 'additional limb.' In 1995, formation of an additional limb was found to be induced by FGF in the chick embryo. This finding subsequently led to the recent understanding of how the limb bud is initially formed, how the limb position is determined, and how the limb identity is determined. Thus, the additional limb has been recognized as a useful experimental system for the study of limb development and its relation to the regionalization of the body. Furthermore, since limb muscles are formed from cells which have migrated from somites and innervation to them rakes place from the spinal cord, the additional limb would also be a powerful tool with which to study the relation of limb morphogenesis to developmental processes of the spinal cord and somites. This review consists of five sections, (1) 'Introduction,' (2) 'How to make additional limbs,' (3) 'Characteristics of the additional limb,' (4) 'Studies with the additional limb,' and (5) 'Concluding remarks.' In the second section, techniques to make additional limbs are reviewed, showing that additional limbs can be made by fairly easy manipulation of the chick embryo. In the third section, the characteristics analyzed so far of the additional limb are summarized, focusing on its morphology. In the fourth section, recent studies on the use of the additional limb are reviewed: experiments on the additional limb have been performed to elucidate the mechanisms governing determination of limb identity by Hox codes and the Tbx family and initiation of limb formation by FGF10. In addition, the roles of SF/HGF in the formation of limb muscles have also been investigated using the additional limb. In the near future, the additional limb will be also used in the study of innervation from the spinal cord, and probably migration of neural crest cells.

    DOI: 10.1007/s004410051265

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  • Bone morphogenetic protein signaling is required for maintenance of differentiated phenotype, control of proliferation, and hypertrophy in chondrocytes.

    Enomoto-Iwamoto Motomi, Iwamoto Masahiro, Mukudai Yoshiki, Kawakami Yasuhiko, Nohno Tsutomu, Higuchi Yoshinobu, Takemoto Seiji, Ohuchi Hideyo, Noji Sumihare, Kurisu Kojiro

    The Journal of cell biology140 ( 2 ) 409 - 418   1998年1月

  • Differential expression of the two closely related LIM-class homeobox genes LH-2A and LH-2B during limb development.

    Nohno Tsutomu, Kawakami Yasuhiko, Wada Naoyuki, Ishikawa Tetsuya, Ohuchi Hideyo, Noji Sumihare

    Biochemical and biophysical research communications238 ( 2 ) 506 - 511   1997年9月

  • A chick wingless mutation causes abnormality in maintenance of Fgf8 expression in the wing apical ridge, resulting in loss of the dorsoventral boundary

    Ohuchi Hideyo, Shibusawa Mami, Nakagawa Takashi, Ohata Takeshi, Yoshioka Hidefumi, Hirai Yasokazu, Nohno Tsutomu, Noji Sumihare, Kondo Norio

    Mechanisms of development62 ( 1 ) 3 - 13   1997年2月

  • New gene, nel, encoding a Mr 91 K protein with EGF-like repeats is strongly expressed in neural tissues of early stage chick embryos (vol 203, pg 202, 1995)

    S Matsuhashi, S Noji, E Koyama, F Myokai, H Ohuchi, S Taniguchi, K Hori

    DEVELOPMENTAL DYNAMICS207 ( 2 ) 233 - 234   1996年10月

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    記述言語:英語   出版者・発行元:WILEY-LISS  

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  • Polarizing activity, Sonic hedgehog, and tooth development in embryonic and postnatal mouse.

    Koyama Eiki, Yamaai Tomoichiro, Iseki Sachiko, Ohuchi Hideyo, Nohno Tsutomu, Yoshioka Hidefumi, Hayashi Yoshio, Leatherman Judith L, Golden Eleanor B, Noji Sumihare, Pacifici Maurizio

    Developmental dynamics : an official publication of the American Association of Anatomists206 ( 1 ) 59 - 72   1996年5月

  • Sonic hedgehog is expressed in epithelial cells during development of whisker, hair, and tooth.

    Iseki Sachiko, Araga Akihiro, Ohuchi Hideyo, Nohno Tsutomu, Yoshioka Hidefumi, Hayashi Fumio, Noji Sumihare

    Biochemical and biophysical research communications218 ( 3 ) 688 - 693   1996年1月

  • A Drosophila receptor-type tyrosine kinase (DFR1) acts as a fibroblast growth factor receptor in Xenopus embryos

    Hidefumi Yoshioka, Hideyo Ohuchi, Yoshiyasu Ishimaru, Tetsuya Ishikawa, Tsutomu Nohno, Kaoru Saigo, Sumihare Noji

    Development Growth and Differentiation38 ( 6 ) 617 - 624   1996年

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    記述言語:英語   出版者・発行元:Blackwell Publishing  

    Members of the fibroblast growth factor (FGF) family play important roles in various developmental processes in vertebrates. Since two genes closely related to the vertebrate FGF receptor (FGFR) genes DFR1 and DFR2/breathless have already been reported in Drosophila, the existence of a Drosophila FGF has been predicted. In the present study, we examined whether DFR1 is functionally interchangeable with a vertebrate FGFR in the Xenopus system. First, we found that the expression of DFR1 promoted Ca2+ efflux in response to human basic (b)FGF in Xenopus oocytes, whereas the coexpression of a dominant negative form of DFR1 (ΔDFR1) with a chick FGFR1/cek1 inhibited promotion of Ca2+ efflux induced by the expression of cek1 in the oocyte. Second, the expression of ΔDFR1 was observed to induce a defect in the posterior structure of the Xenopus embryo at stage 30, as observed with a dominant negative form of cek1 (Δcek1). Third, we found that the expression of ΔDFR1 inhibited the expression of FGF-regulated genes such as Xbra, Xnot, and Xshh in Xenopus embryos at stage 11, while the coexpression of DFR1 with ΔDFR1 could rescue the inhibited expression of FGF-regulated genes. These results indicate that DFR1 acts as an FGFR in Xenopus embryos and that an FGF is likely to exist in Drosophila.

    DOI: 10.1046/j.1440-169X.1996.t01-5-00005.x

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  • An additional limb can be induced from the flank of the chick embryo by FGF4

    Hideyo Ohuchi, Takashi Nakagawa, Masaki Yamauchi, Takeshi Ohata, Hidefumi Yoshioka, Takashi Kuwana, Tatsuo Mima, Takashi Mikawa, Tsutumu Nohno, Sumihare Noji

    Biochemical and Biophysical Research Communications209 ( 3 ) 809 - 816   1995年4月

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    記述言語:英語  

    To elucidate what initiates formation of the limb, we have attempted to induce an additional limb from the flank of the chick embryo by infecting retrovirus or implanting cells. We report here that an additional limb can be formed from the flank when we implant fibroblast growth factor 4 (Fgf4)-expressing cells into the lateral plate mesoderm at the pre-limb bud stage. In a newly formed limb bud, expressions of both Sonic hedgehog and chick Fgf4, which are authentic morphogenetic signals from the zone of polarizing activity and the apical ectodermal ridge, respectively, are induced by the implanted cells. Thus, it is concluded that the competence for limb development is present along the flank of the chick embryo and that FGF4 applied ectopically at the pre-limb bud stage can alter the developmental fate of flank cells to become limb cells. The present experimental system will contribute to a further elucidation on how the limb is formed. © 1995 by Academic Press, Inc.

    DOI: 10.1006/bbrc.1995.1572

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  • Involvement of the Sonic Hedgehog gene in chick feather formation

    Tsutomu Nohno, Yasuhiko Kawakami, Hideyo Ohuchi, Akira Fujiwara, Hidefumi Yoshioka, Sumihare Noji

    Biochemical and Biophysical Research Communications206 ( 1 ) 33 - 39   1995年1月

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    記述言語:英語  

    To elucidate the molecular mechanisms of chick feather formation, we observed expression patterns of the Sonic hedgehog (Shh) gene, which is one of the vertebrate homologs of the Drosophila segment polarity gene, hedgehog, and encodes a signaling molecule functioning in limb pattern formation and motor neuron induction. We found that the Shh gene is also expressed in the apical region of the feather placodes and then in nine to eleven longitudinal stripes along feather filaments. The stripe was found to correspond to one of the outer marginal zones of each barb ridge, termed the zone of Shh expression. No significant expression signal was detected in the scale bud of developing legs. Thus, Shh is likely to function as an epithelial signaling molecule in epitheliomesenchymal interaction during feather formation. Furthermore, since genes of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-4 (FGF-4) are coexpressed with Shh during feather formation as observed in limb morphogenesis, interactions among FGF-4, Shh and BMP-2 may be involved in formation of feather filaments and barbs in a similar fashion as elucidated in limb pattern formation. © 1995 Academic Press. Inc.

    DOI: 10.1006/bbrc.1995.1005

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  • A Chicken WNT Gene, WNT-11, Is Involved in Dermal Development

    Nohuya Tanda, Hidcyo Ohuchi, Hidefumi Yoshioka, Sumiharc Noji, Tsutomu Nohno

    Biochemical and Biophysical Research Communications211 ( 1 ) 123 - 129   1995年

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    記述言語:英語  

    We have isolated a new member of the Wnt gene family, Wnt-11, from chick embryo cDNA library and examined the expression pattern during embryogenesis by in situ hybridization. The Wnt-11 gene encodes Cys-rich secretory protein distantly related to Wnt-1 through Wnt-8 from the mouse and Xenopus. Expression of the Wnt-11 gene became evident at stage 14 in the dorsolateral region of somites and gradually restricted to the dermatome at stage 19 and later. In contrast to the other Wnt genes, Wnt-11 was not expressed in the neuroepithelium throughout stages 14-26. At stage 24 and later, Wnt-11 was expressed in the subectodermal mesenchyme of the limb and feather buds. The unique expression pattern of Wnt-11 in the paraxial mesoderm and dermatome suggests that Wnt-11 may play an important role in dermal development. © 1995 Academic Press.

    DOI: 10.1006/bbrc.1995.1786

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  • Truncated Type II Receptor for BMP-4 Induces Secondary Axial Structures in Xenopus Embryos

    Tetsuya Ishikawa, Hidefumi Yoshioka, Hidcyo Ohuchi, Sumihare Noji, Tsutomu Nohno

    Biochemical and Biophysical Research Communications216 ( 1 ) 26 - 33   1995年

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    記述言語:英語  

    BRK-3 is a vertebrate type II receptor for BMP-4 distantly related to invertebrate type II receptors for BMP-2/BMP-4/dpp, such as daf-d, and punt. BRK-3 has a long carboxy-terminal sequence following intracellular kinase domain and is capable of forming a high-affinity complex with a type I receptor, BRK-2. To examine the role of BRK-2 + BRK-3 receptor complex in BMP signaling during early embryogenesis, the dominant-negative form of BRK-3 was ectopically expressed in the Xenopus embryos. A secondary body axis expressing the Sonic hedgehog and N-CAM genes is induced by injecting mRNA encoding truncated form of BRK-3 into ventral marginal region, implicating the BMP signaling in axial mesoderm induction. Formation of the secondary axis depends on whether the deletion extends into the kinase domain, not into the carboxy-terminal tail, suggesting that the kinase domain, but not the tail region, is essential for BMP signaling. © 1995 by Academic Press, Inc.

    DOI: 10.1006/bbrc.1995.2587

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  • Involvement of androgen-induced growth factor (FGF-8) gene in mouse embryogenesis and morphogenesis

    Hideyo Ohuchi, Hidefumi Yoshioka, Akira Tanaka, Yasuhiko Kawakami, Tsutomu Nohno, Sumihare Noji

    Biochemical and Biophysical Research Communications204 ( 2 ) 882 - 888   1994年10月

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    記述言語:英語  

    Androgen-induced growth factor (AIGF/FGF-8) has purified from a mouse mammary carcinoma cell line and its cDNA cloning revealed that AIGF is the eighth member of the fibroblast growth factor family. To clarify a biological role of FGF-8, Northern blot analysis of various adult tissues was carried out, but no expression was found. However, whole mount in situ hybridization of 7.5- to 14.5-day gestation (E7.5-14.5) embryos revealed six predominant expression domains: [1]primitive streak region (E7.5-9.75)
    [2]midbrain-hindbrain border (E8.0-10.5)
    [3]rostral forebrain (E8.0-10.5)
    [4]limb ectoderm and apical ectodermal ridge (E9.25-13.5)
    [5]nasal placode and epithelium (E9.5-12.5)
    and [6]branchial arch ectoderm (E8.5-11.5). The embryonic expression pattern suggests a unique role of FGF-8 in mouse development, especially in gastrulation, brain development, and limb and facial morphogenesis. © 1994 Academic Press. All rights reserved.

    DOI: 10.1006/bbrc.1994.2542

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  • Regional expression of the Cwnt-4 Gene in developing chick central nervous system in relationship to the diencephalic neuromere D2 and a dorsal domain of the spinal-cord

    Hidefumi Yoshioka, Hideyo Ohuchi, Tsutomu Nohno, Akira Fujiwara, Nobuya Tanda, Yasuhiko Kawakami, Sumihare Noji

    Biochemical and Biophysical Research Communications203 ( 3 ) 1581 - 1588   1994年9月

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    記述言語:英語  

    Members of the Wnt gene family code for cysteine-rich, secreted proteins, which are differentially expressed in the developing brain and possibly act as an intercellular signaling molecule. A Wnt gene, e.g., Wnt-1 is known to be essential for specification of the midbrain cell fate. Since other members of the Wnt genes are likely to be also involved in development of the brain, we searched chick Wnt members expressing in specific domains of the brain. We found several novel Wnt members expressing specifically in the central nervous system of chick embryos. One of the genes was intensely expressed in a segment of the diencephalon at stages 14 and 20 and in a dorsal region of the spinal cord at stage 20. Although the expression patterns differ from those of mouse or Xenopus Wnt-4, the gene is highly homologous to the mouse or frog Wnt-4. Thus, we designated the gene as Cwnt-4 (chick Wnt-4). Our results suggested that Cwnt-4 is involved in segmentation of the forebrain into the neuromere D2 and in differentiation of the dorsal region of the spinal cord. © 1994 Academic Press, Inc.

    DOI: 10.1006/bbrc.1994.2367

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  • Expression patterns of two fibroblast growth factor receptor genes during early chick eye development.

    Ohuchi Hideyo, Koyama Eiki, Myokai Fumio, Nohno Tsutomu, Shiraga Fumio, Matsuo Toshihiko, Matsuo Nobuhiko, Taniguchi Shigehiko, Noji Sumihare

    Experimental eye research58 ( 6 ) 649 - 658   1994年6月

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  • A chicken homeobox gene related to drosophila paired is predominantly expressed in the developing limb

    Tsutomu Nohno, Eiki Koyama, Fumio Myokai, Shigehiko Taniguchi, Hideyo Ohuchi, Taiichi Saito, Sumihare Noji

    Developmental Biology158 ( 1 ) 254 - 264   1993年

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    記述言語:英語  

    We identified a homeobox-containing gene, Prx-1, isolated from the chick limb bud cDNA library. The homeodomain sequence is related to Drosophila paired and gooseberry and mouse Pax-3, Pax-6, and Pax-7. The deduced amino acid sequence of the Prx-1 gene product reveals the absence of a paired-box sequence and extensive similarity to mouse S8 and MHox homeodomain proteins, thus constituting a new class of homeobox gene. Using an in situ hybridization method, the Prx-1 gene is shown to be expressed predominantly in the limb bud and visceral arches. At early stages of limb development, distal mesodertnal cells express the homeobox gene with an apparent gradient along the proximal-distal axis. The signal is absent in the apical and nonridge ectoderm. Removal of the apical ectodermal ridge had no apparent effect on the subsequent expression of Prx-1 in the limb mesenchyme. The Prx-1-expressing cells are later confined to the interdigital and perichondrial regions. The Prx-1 transcripts are also detectable in the mesenchyme of the visceral arches and facial primordia subjacent to the ectoderm. The Prx-1 gene is weakly expressed in somites and condensing vertebrae. No signal is detectable in neural tube and ectodermal epithelium. These results suggest that the Prx-1 homeodomain protein is involved in the differentiation of bone, muscle, and other tissues of mesodermal origin during limb development. © 1993 Academic Press, Inc. © 1993 by Academic Press, Inc.

    DOI: 10.1006/dbio.1993.1184

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  • EXPRESSION PATTERN OF THE ACTIVIN RECEPTOR TYPE-IIA GENE DURING DIFFERENTIATION OF CHICK NEURAL TISSUES, MUSCLE AND SKIN

    H OHUCHI, S NOJI, E KOYAMA, F MYOKAI, K NISHIKAWA, T NOHNO, K TASHIRO, K SHIOKAWA, N MATSUO, S TANIGUCHI

    FEBS LETTERS303 ( 2-3 ) 185 - 189   1992年6月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    To elucidate target cells of activins during embryogenesis we isolated cDNAs of chick activin receptor type II (cActR-II) and studied expression patterns of the cActR-II gene by in situ hybridization. Transcripts of cActR-II were observed in neuroectoderm developing to spinal cord, brain and eyes, in surface ectoderm differentiating to epidermis, and in myotomes differentiating to muscles. The expression patterns of cActR-II suggest that activin and its receptor are involved in differentiation of chick neural tissues, muscle and skin after inducing the dorsal mesoderm.

    DOI: 10.1016/0014-5793(92)80515-I

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  • EXPRESSION PATTERNS OF FGF AND ACTIVIN RECEPTORS IN DEVELOPING CHICK EYE - AS REVEALED BY INSITU HYBRIDIZATION

    H OHUCHI, S NOJI, T NOHNO, E KOYAMA, F MYOKAI, K NISHIKAWA, S TANIGUCHI, N MATSUO

    INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE33 ( 4 ) 828 - 828   1992年3月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:LIPPINCOTT-RAVEN PUBL  

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講演・口頭発表等

  • 神経、内分泌系に発現する光受容タンパク質の分子組織学的解析 招待

    大内淑代, 佐藤恵太

    異分野融合による次世代光生物学研究会  2019年11月7日 

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    記述言語:日本語   会議種別:口頭発表(招待・特別)  

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  • 非視覚性光受容タンパク質の機能解明と モデル動物を用いた眼疾患研究 招待

    大内淑代

    第22回眼科分子生物学研究会  2018年3月11日 

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    記述言語:日本語   会議種別:口頭発表(招待・特別)  

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  • REIC/DKK3 Expression in Mouse Normal Tissues

    大内淑代

    第4回東アジア遺伝子治療推進懇話会  2015年6月21日 

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    記述言語:英語   会議種別:シンポジウム・ワークショップ パネル(指名)  

    開催地:Okayama, Japan  

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共同研究・競争的資金等の研究

  • 眼球伸長におけるFGF10と紫光Opsin5システムの機能解明

    研究課題/領域番号:20K21655  2020年07月 - 2023年03月

    日本学術振興会  科学研究費助成事業 挑戦的研究(萌芽)  挑戦的研究(萌芽)

    大内 淑代, 藤田 洋史, 佐藤 恵太

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    配分額:6370000円 ( 直接経費:4900000円 、 間接経費:1470000円 )

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  • ゲノム編集技術を用いた網膜における長寿遺伝子の機能解明:加齢黄斑変性の予防戦略

    研究課題/領域番号:18K09410  2018年04月 - 2021年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    森實 祐基, 大内 淑代, 米澤 朋子

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    配分額:4420000円 ( 直接経費:3400000円 、 間接経費:1020000円 )

    加齢黄斑変性(Age-related macular degeneration,以下AMD)は失明に至る難治性の眼疾患であり,加齢を背景とした網膜色素上皮細胞(以下RPE)の機能低下(老化)が原因で発症する。申請者はこれまでに,長寿遺伝子の一つであるAMP活性化プロテインキナーゼ(以下AMPK)に着目し,その活性を薬剤によって変化させ、AMPKの活性化が様々な眼疾患の病態を改善することを明らかにしてきた。しかし,AMPKやその他の長寿遺伝子が網膜やRPEの老化に及ぼす影響は不明であり,また、薬剤の非特異的作用のために,長寿遺伝子の真の機能を検討することが困難であるという問題があった。そこで本研究では,主要な長寿遺伝子であるサーチュイン(以下SIRT)とAMPKに着目し,これらの遺伝子が網膜やRPEの老化に及ぼす影響を,ゲノム編集技術を用いて作成した遺伝子改変RPEおよびマウスを用いて明らかにする。そして,十分な治療法が存在しない加齢黄斑変性の新規予防法を開発するための基盤となる成果を得る。
    上記の目的を達成するために当該年度は、SIRT遺伝子に着目し、CRISPR/Cas9等のゲノム編集技術を用いてiPS細胞のSIRT遺伝子のゲノム編集に着手した。iPS細胞のゲノム編集の前段階として、ゲノム編集の効果を確認するために、培養が簡便でゲノム編集を高い確率で行うことができる細胞を用いてゲノム編集を行った。

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担当授業科目

  • 人体の構造:入門 (2020年度) 特別  - その他

  • 人体構造学 (2020年度) 集中  - その他

  • 医学セミナー(テュートリアル) (2020年度) 第1学期  - 火2,火3

  • 医学セミナー(チュートリアル) (2020年度) 第1学期  - 火2,火3

  • 発生学 (2020年度) 特別  - その他

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  • 細胞組織学I(演習・実習) (2020年度) 特別  - その他

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  • 細胞組織工学演習 (2020年度) 通年  - その他

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