Updated on 2024/02/02

写真a

 
MOMOTA Ryusuke
 
Organization
Faculty of Medicine, Dentistry and Pharmaceutical Sciences Assistant Professor
Position
Assistant Professor
Profile
Specialized in extracellular matrices and related human diseases. Teaching human anatomy. Passionate about new technologies. Writing codes in Python.
External link

Degree

  • PhD ( Okayama University )

Research Interests

  • Mitochondria

  • Anatomy

  • Collagen

  • Extracellular matrix

  • Basement membrane

Research Areas

  • Life Science / Nutrition science and health science  / 加齢による筋・上皮組織の変化

  • Life Science / Anatomy

  • Informatics / Database  / Development of 3D Anatomy Viewer

  • Life Science / Developmental biology  / Roles of extracellular matrices during embryogenesis

  • Nanotechnology/Materials / Chemistry and chemical methodology of biomolecules  / Basement membrane collagen synthesis

  • Informatics / Life, health and medical informatics  / Integration of anatomical terms

▼display all

Education

  • The University of Tokyo   教養学部   基礎科学科

      More details

    Country: Japan

    Notes: 第1

    researchmap

  • The University of Tokyo   大学院総合文化研究科  

      More details

Professional Memberships

 

Papers

  • 長鎖型XVIII型コラーゲンは新規膜結合型コラーゲンである可能性が高い

    上野 智規, 米澤 朋子, 百田 龍輔, 佐々木 隆子, 大橋 俊孝, 水野 一乘

    日本結合組織学会学術大会プログラム・抄録集   55回   147 - 147   2023.6

     More details

    Language:Japanese   Publisher:日本結合組織学会  

    researchmap

  • XVIII型コラーゲン欠損マウスにおける皮膚創傷治癒の解析

    米澤 朋子, 前川 明日華, 前場 崇宏, 百田 龍輔, 渋谷 千晶, 岩田 宗一郎, 大野 充昭, 大橋 俊孝

    日本結合組織学会学術大会プログラム・抄録集   54回   119 - 119   2022.6

     More details

    Language:Japanese   Publisher:日本結合組織学会  

    researchmap

  • Lack of collagen α6(IV) chain in mice does not cause severe-to-profound hearing loss or cochlear malformation, a distinct phenotype from nonsyndromic hearing loss with COL4A6 missense mutation. International journal

    Shaoying Tang, Tomoko Yonezawa, Yukihide Maeda, Mitsuaki Ono, Takahiro Maeba, Toru Miyoshi, Ryusuke Momota, Yasuko Tomono, Toshitaka Oohashi

    PloS one   16 ( 4 )   e0249909   2021

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Congenital hearing loss affects 1 in every 1000 births, with genetic mutations contributing to more than 50% of all cases. X-linked nonsyndromic hereditary hearing loss is associated with six loci (DFNX1-6) and five genes. Recently, the missense mutation (c.1771G>A, p.Gly591Ser) in COL4A6, encoding the basement membrane (BM) collagen α6(IV) chain, was shown to be associated with X-linked congenital nonsyndromic hearing loss with cochlear malformation. However, the mechanism by which the COL4A6 mutation impacts hereditary hearing loss has not yet been elucidated. Herein, we investigated Col4a6 knockout (KO) effects on hearing function and cochlear formation in mice. Immunohistochemistry showed that the collagen α6(IV) chain was distributed throughout the mouse cochlea within subepithelial BMs underlying the interdental cells, inner sulcus cells, basilar membrane, outer sulcus cells, root cells, Reissner's membrane, and perivascular BMs in the spiral limbus, spiral ligament, and stria vascularis. However, the click-evoked auditory brainstem response analysis did not show significant changes in the hearing threshold of Col4a6 KO mice compared with wild-type (WT) mice with the same genetic background. In addition, the cochlear structures of Col4a6 KO mice did not exhibit morphological alterations, according to the results of high-resolution micro-computed tomography and histology. Hence, loss of Col4a6 gene expression in mice showed normal click ABR thresholds and normal cochlear formation, which differs from humans with the COL4A6 missense mutation c.1771G>A, p.Gly591Ser. Therefore, the deleterious effects in the auditory system caused by the missense mutation in COL4A6 are likely due to the dominant-negative effects of the α6(IV) chain and/or α5α6α5(IV) heterotrimer with an aberrant structure that would not occur in cases with loss of gene expression.

    DOI: 10.1371/journal.pone.0249909

    PubMed

    researchmap

  • コラーゲン研究の新展開:基礎から創薬まで マウス皮膚創傷モデルにおけるXVIII型コラーゲンの解析

    米澤 朋子, 前場 崇宏, Tang Shaoying, 大野 充昭, 百田 龍輔, 稲川 喜一, 大橋 俊孝

    日本生化学会大会プログラム・講演要旨集   93回   [1S08e - 06]   2020.9

     More details

    Language:Japanese   Publisher:(公社)日本生化学会  

    researchmap

  • Prolonged Tachycardia with Higher Heart Rate Is Associated with Higher ICU and In-hospital Mortality. Reviewed

    Hayashi M, Taniguchi A, Kaku R, Fujimoto S, Isoyama S, Manabe S, Yoshida T, Suzuki S, Shimizu K, Morimatsu H, Momota R

    Acta medica Okayama   73 ( 2 )   147 - 153   2019.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Tachycardia is common in intensive care units (ICUs). It is unknown whether tachycardia or prolonged tachycardia affects patient outcomes. We investigated the association between tachycardia and mortality in critically ill patients. This retrospective cohort study's primary outcome was patient mortality in the ICU and the hospital. We stratified the patients (n=476) by heart rate (HR) as LowHR, MediumHR, and HighHR groups. We also stratified them by their durations of HR >100 (prolonged HR; tachycardia): MildT, ModerateT, and SevereT groups. We determined the six groups' mortality. The ICU mortality rates of the LowHR, MediumHR, and HighHR groups were 1.0%, 1.5%, and 7.9%, respectively; significantly higher in the HighHR vs. LowHR group. The in-hospital mortality rates of these groups were 1%, 4.5%, and 14.6%, respectively; significantly higher in the HighHR vs. LowHR group. The ICU mortality rates of the MildT, ModerateT, and SevereT groups were 0.9%, 5.6%, and 57.1%, respectively. The mortality of the HRT=0 (i.e., all HR ≤ 100) patients was 0%. The in-hospital mortality rates of the MildT, ModerateT, and SevereT groups were 1.8%, 16.7%, and 85.7%, respectively; that of the HRT=0 patients was 0.5%. Both higher HR and prolonged tachycardia were associated with poor outcomes.

    DOI: 10.18926/AMO/56650

    PubMed

    researchmap

  • Unripe peach (Prunus persica) extract ameliorates damage from UV irradiation and improved collagen XVIII expression in 3D skin model. Reviewed International journal

    Tomoko Yonezawa, Ryusuke Momota, Hideki Iwano, Steven Zhao, Tomohiro Hakozaki, Chieko Soh, Shigetoyo Sawaki, Kazumi Toyama, Toshitaka Oohashi

    Journal of cosmetic dermatology   2018.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    INTRODUCTION: Collagen type XVIII regulates cellular activities of adjacent cells at the dermal-epidermal junction (DEJ). To investigate its possible changes during aging, we compared its mRNA levels and protein localization in skin samples from female participants aged 20-70 years old. In addition, we evaluated the beneficial effects of unripe peach extracts in a 3D skin model. METHODS: Sun-exposed or sun-protected female skin samples were compared by DNA array or by immunohistochemistry for basement membrane components. To evaluate protective effects of fresh unripe peach extract, UV-B irradiated human 3D skin models were incubated in the presence or absence of the extract, followed by measurements of mRNA levels by real-time PCR, or by immunohistochemistry. RESULTS: In aged skin samples, COL18A1 mRNA levels were lower and the protein localization exhibited less intensive signal by anti-collagen type XVIII immunostaining. As observed in the skin tissues, collagen type XVIII exists at the DEJ in the 3D skin model. Fresh unripe peach extract significantly improved mRNA levels and partially localizations of collagen type XVIII, suggesting that fresh unripe peach extract ameliorates DEJ damages caused by UV-B irradiation. CONCLUSION: Collagen type XVIII and fresh unripe peach extract can be promising protective cosmetic strategies against skin aging.

    DOI: 10.1111/jocd.12841

    PubMed

    researchmap

  • Network of anatomical texts (NAnaTex), an open-source project for visualizing the interaction between anatomical terms Reviewed

    Ryusuke Momota, Aiji Ohtsuka

    Anatomical Science International   93 ( 1 )   149 - 153   2018.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Tokyo  

    Anatomy is the science and art of understanding the structure of the body and its components in relation to the functions of the whole-body system. Medicine is based on a deep understanding of anatomy, but quite a few introductory-level learners are overwhelmed by the sheer amount of anatomical terminology that must be understood, so they regard anatomy as a dull and dense subject. To help them learn anatomical terms in a more contextual way, we started a new open-source project, the Network of Anatomical Texts (NAnaTex), which visualizes relationships of body components by integrating text-based anatomical information using Cytoscape, a network visualization software platform. Here, we present a network of bones and muscles produced from literature descriptions. As this network is primarily text-based and does not require any programming knowledge, it is easy to implement new functions or provide extra information by making changes to the original text files. To facilitate collaborations, we deposited the source code files for the network into the GitHub repository (https://github.com/ryusukemomota/nanatex) so that anybody can participate in the evolution of the network and use it for their own non-profit purposes. This project should help not only introductory-level learners but also professional medical practitioners, who could use it as a quick reference.

    DOI: 10.1007/s12565-017-0410-1

    Scopus

    PubMed

    researchmap

  • Type IV collagen α6 chain is a regulator of keratin 10 in keratinization of oral mucosal epithelium Reviewed International journal

    Komori, T., Ono, M., Hara, E.S., Ueda, J., Nguyen, H.T.T., Nguyen, H.T., Yonezawa, T., Maeba, T., Kimura-Ono, A., Takarada, T., Momota, R., Maekawa, K., Kuboki, T., Oohashi, T.

    Scientific Reports   8 ( 1 )   2612 - 2612   2018

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Keratinized mucosa is of fundamental importance to maintain healthy gingival tissue, and understanding the mechanisms of oral mucosa keratinization is crucial to successfully manage healthy gingiva. Previous studies have shown a strong involvement of the basement membrane in the proliferation and differentiation of epithelial cells. Therefore, first, to identify the keratinized mucosa-specific basement membrane components, immunohistochemical analysis for the six alpha chains of type IV collagen was performed in 8-week-old mice. No difference in the expression pattern of type IV collagen α1(IV) and α2(IV) chains was observed in the keratinized and non-keratinized mucosa. Interestingly, however, type IV collagen α5(IV) and α6(IV) chains specifically were strongly detected in the keratinized mucosa. To analyze the functional roles of the type IV collagen isoform α6(IV) in oral mucosa keratinization, we analyzed Col4a6-knockout mice. Epithelial developmental delay and low levels of KRT10 were observed in new-born Col4a6-knockout mice. Additionally, in vitro experiments with loss-of function analysis using human gingival epithelial cells confirmed the important role of α6(IV) chain in epithelial keratinization. These findings indicate that α112:α556 (IV) network, which is the only network that includes the α6(IV) chain, is one regulator of KRT10 expression in keratinization of oral mucosal epithelium.

    DOI: 10.1038/s41598-018-21000-0

    Scopus

    PubMed

    researchmap

  • Effects of aerobic exercise on capillary architecture of extensor digitorum Longus in Rat Reviewed

    Shinichiro Murakami, Masahiro Sakita, Hiroyo Kondo, Miho Kanazashi, Masayuki Tanaka, Minoru Tanaka, Aiji Ohtuka, Ryusuke Momota, Hidemi Fujino

    FASEB JOURNAL   28 ( 1 )   2014.4

     More details

    Language:English   Publisher:FEDERATION AMER SOC EXP BIOL  

    Web of Science

    researchmap

  • 致死となるマルチプレキシン(XV/XVIII型コラーゲン)変異ショウジョウバエの解析

    百田 龍輔, 楢崎 正博, 小見山 高明, 内藤 一郎, 二宮 善文, 大塚 愛二

    日本結合組織学会学術大会・マトリックス研究会大会合同学術集会プログラム・抄録集   45回・60回   86 - 86   2013.6

     More details

    Language:Japanese   Publisher:日本結合組織学会・マトリックス研究会  

    researchmap

  • Drosophila type XV/XVIII collagen mutants manifest integrin mediated mitochondrial dysfunction, which is improved by cyclosporin A and losartan Reviewed International journal

    Ryusuke Momota, Masahiro Narasaki, Takaaki Komiyama, Ichiro Naito, Yoshifumi Ninomiya, Aiji Ohtsuka

    INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY   45 ( 5 )   1003 - 1011   2013.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PERGAMON-ELSEVIER SCIENCE LTD  

    Vertebrate collagen types XV and XVIII are broadly distributed basement membrane components, classified into a structurally distinct subgroup called "multiplexin collagens". Mutations in mammalian multiplexins are identified in some degenerative diseases such as Knobloch syndrome 1 (KNO1) or skeletal/cardiac myopathies, however, these progressive properties have not been elucidated. Here we investigated Drosophila mutants of Multiplexin (Mp), the only orthologue of vertebrate collagen types XV and XVIII, to understand the pathogenesis of multiplexin-related diseases. The mp mutants exhibited morphological changes in cardiomyocytes and progressive dysfunction of the skeletal muscles, reminiscent phenotypes observed in Col15a1-null mice. Ultrastructural analysis revealed morphologically altered mitochondria in mutants' indirect flight muscles (IFMs), resulting in severely attenuated ATP production and enhanced reactive oxygen species (ROS) production. In addition, mutants' IFMs exhibited diminished PPS integrin clustering and abolished focal adhesion kinase (FAK) phosphorylation. Furthermore, mutants' defective IFMs are improved by the administrations of cyclosporin A, an inhibitor against mitochondrial permeability transition pore (mPTP) opening or losartan, an angiotensin II type 1 receptor (AT1R) blocker. Thus, our results suggest that Mp modulates mPTP opening and AT1R activity through its binding to integrin and that lack of Mp causes unregulated mPTP opening and AT1R activity, leading to mitochondrial dysfunctions. Hence, our results provide new insights towards the roles of multiplexin collagens in mitochondrial homeostasis and may serve as pharmacological evidences for the potential use of cyclosporin A or losartan for the therapeutic strategies. (C) 2013 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.biocel.2013.02.001

    Web of Science

    PubMed

    researchmap

  • Architecture of the subendothelial elastic fibers of small blood vessels and variations in vascular type and size Reviewed

    Akira Shinaoka, Ryusuke Momota, Eri Shiratsuchi, Mitsuko Kosaka, Kanae Kumagishi, Ryuichi Nakahara, Ichiro Naito, Aiji Ohtsuka

    Microscopy and Microanalysis   19 ( 2 )   406 - 414   2013.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Most blood vessels contain elastin that provides the vessels with the resilience and flexibility necessary to control hemodynamics. Pathophysiological hemodynamic changes affect the remodeling of elastic components, but little is known about their structural properties. The present study was designed to elucidate, in detail, the three-dimensional (3D) architecture of delicate elastic fibers in small vessels, and to reveal their architectural pattern in a rat model. The fine vascular elastic components were observed by a newly developed scanning electron microscopy technique using a formic acid digestion with vascular casts. This method successfully visualized the 3D architecture of elastic fibers in small blood vessels, even arterioles and venules. The subendothelial elastic fibers in such small vessels assemble into a sheet of meshwork running longitudinally, while larger vessels have a higher density of mesh and thicker mesh fibers. The quantitative analysis revealed that arterioles had a wider range of mesh density than venules
    the ratio of density to vessel size was higher than that in venules. The new method was useful for evaluating the subendothelial elastic fibers of small vessels and for demonstrating differences in the architecture of different types of vessels. © Microscopy Society of America 2013.

    DOI: 10.1017/S1431927612014341

    Scopus

    PubMed

    researchmap

  • XV/XVIII型コラーゲンの新たな生物学的機能の探索

    百田 龍輔, 内藤 一郎, 二宮 善文, 大塚 愛二

    解剖学雑誌   87 ( 2 )   44 - 44   2012.6

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • Abnormalities in the Fiber Composition and Capillary Architecture in the Soleus Muscle of Type 2 Diabetic Goto-Kakizaki Rats Reviewed

    Shinichiro Murakami, Naoto Fujita, Hiroyo Kondo, Isao Takeda, Ryusuke Momota, Aiji Ohtsuka, Hidemi Fujino

    SCIENTIFIC WORLD JOURNAL   2012   680189   2012

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:HINDAWI PUBLISHING CORPORATION  

    Type 2 diabetes mellitus is linked to impaired skeletal muscle glucose uptake and storage. This study aimed to investigate the fiber type distributions and the three-dimensional (3D) architecture of the capillary network in the skeletal muscles of type 2 diabetic rats. Muscle fiber type transformation, succinate dehydrogenase (SDH) activity, capillary density, and 3D architecture of the capillary network in the soleus muscle were determined in 36-week-old Goto-Kakizaki (GK) rats as an animal model of nonobese type 2 diabetes and age-matched Wistar (Cont) rats. Although the soleus muscle of Cont rats comprised both type I and type IIA fibers, the soleus muscle of GK rats had only type I fibers. In addition, total SDH activity in the soleus muscle of GK rats was significantly lower than that in Cont rats because GK rats had no high-SDH activity type IIA fiber in the soleus muscle. Furthermore, the capillary diameter, capillary tortuosity, and microvessel volume in GK rats were significantly lower than those in Cont rats. These results indicate that non-obese diabetic GK rats have muscle fiber type transformation, low SDH activity, and reduced skeletal muscle capillary content, which may be related to the impaired glucose metabolism characteristic of type 2 diabetes.

    DOI: 10.1100/2012/680189

    Web of Science

    PubMed

    researchmap

  • Drosophila type XV/XVIII collagen, Mp, is involved in Wingless distribution Reviewed International journal

    Ryusuke Momota, Ichiro Naito, Yoshifumi Ninomiya, Aiji Ohtsuka

    MATRIX BIOLOGY   30 ( 4 )   258 - 266   2011.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Multiplexin (Mp) is the Drosophila orthologue of vertebrate collagens XV and XVIII. Like them, Mp is widely distributed in the basement membranes of the developing embryos, including those of neuroblasts in the central and peripheral nervous systems, visceral muscles of the gut, and contractile cardioblasts. Here we report the identification of mutant larvae bearing piguBac transposon insertions that exhibit decrease Mp production associated with abdominal cuticular and wing margin defects, malformation of sensory organs and impaired sensitivity to physical stimuli. Additional findings include the abnormal ultrastructure of fatbody associated with abnormal collagen IV deposition, and reduced Wingless deposition. Collectively, these findings are consistent with the notion that Mp is required for the proper formation and/or maintenance of basement membrane, and that Mp may be involved in establishing the Wingless signaling gradients in the Drosophila embryo. (C) 2011 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.matbio.2011.03.008

    Web of Science

    PubMed

    researchmap

  • 微小血管内皮下弾性線維構造とその血行動態との関係

    品岡 玲, 百田 龍輔, 小阪 美津子, 内藤 一郎, 中原 龍一, 大塚 愛二

    日本結合組織学会学術大会・マトリックス研究会大会合同学術集会プログラム・抄録集   43回・58回   105 - 105   2011.5

     More details

    Language:Japanese   Publisher:日本結合組織学会・マトリックス研究会  

    researchmap

  • ショウジョウバエmultiplexinの生物学的機能

    百田 龍輔, 内藤 一郎, 二宮 善文, 大塚 愛二

    日本結合組織学会学術大会・マトリックス研究会大会合同学術集会プログラム・抄録集   43回・58回   72 - 72   2011.5

     More details

    Language:Japanese   Publisher:日本結合組織学会・マトリックス研究会  

    researchmap

  • II型糖尿病ラット(GOTO-KAKIZAKIラット)のヒラメ筋の筋線維タイプの変化

    熊岸 加苗, 村上 慎一郎, 藤野 英己, 武田 功, 百田 龍輔, 近藤 浩代, 大塚 愛二

    解剖学雑誌   85 ( 2 )   82 - 82   2010.6

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • 共焦点レーザー顕微鏡を用いたラットの遅筋と速筋の毛細血管構築の比較

    村上 慎一郎, 熊岸 加苗, 藤野 英己, 武田 功, 百田 龍輔, 近藤 浩代, 大塚 愛二

    解剖学雑誌   85 ( 2 )   82 - 82   2010.6

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • 非インスリン依存型糖尿病モデルラットのヒラメ筋の毛細血管構築、ミオシン重鎖アイソフォーム、及び酸化的リン酸化活性の変化

    村上 慎一郎, 藤野 英己, 近藤 浩代, 武田 功, 百田 龍輔, 熊岸 加苗, 大塚 愛二

    解剖学雑誌   85 ( Suppl. )   114 - 114   2010.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • 動脈の内弾性板線維配列 とくに断裂部の超微細構造解析

    品岡 玲, 百田 龍輔, 内藤 一郎, 大塚 愛二

    解剖学雑誌   85 ( Suppl. )   200 - 200   2010.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • Comparison of Capillary Architecture between Slow and Fast Muscles in Rats Using a Confocal Laser Scanning Microscope Reviewed

    Shinichiro Murakami, Hidemi Fujino, Isao Takeda, Ryusuke Momota, Kanae Kumagishi, Aiji Ohtsuka

    ACTA MEDICA OKAYAMA   64 ( 1 )   11 - 18   2010.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    The skeletal muscle is classified into 2 types, slow oxidative or fast glycolytic muscle. For further characterization, we investigated the capillary architecture in slow and fast muscles. The rat soleus and extensor digitorum longus (EDL) muscles were used as representatives of slow and fast muscles, respectively. To investigate capillary density, sections of both types of muscle were stained with alkaline phosphatase; the soleus muscle showed more intense reactivity, indicating that it had a denser capillary structure than the EDL muscle. We then injected fluorescent contrast medium into samples of both muscle types for light and confocal-laser microscopic evaluation. The capillary density and capillary-to-fiber ratio were significantly higher, and the course of the capillaries was more tortuous, in the soleus muscle than in the EDL muscle. Capillary coursed more tortuously in the soleus than in the EDL muscle. Succinate dehydrogenase (SDH) activity, an indicator of mitochondrial oxidative capacity, and vascular endothelial growth factor (VEGF) expression were also significantly higher in the soleus muscle. Thus, we conclude that slow oxidative muscle possess a rich capillary structure to provide demanded oxygen, and VEGF might be involved in the formation and/or maintenance of this highly capillarized architecture.

    DOI: 10.18926/AMO/32859

    Web of Science

    PubMed

    researchmap

  • Comparison of Capillary Architecture between Slow and Fast Muscles in Rats Using a Confocal Laser Scanning Microscope Reviewed

    Shinichiro Murakami, Hidemi Fujino, Isao Takeda, Ryusuke Momota, Kanae Kumagishi, Aiji Ohtsuka

    ACTA MEDICA OKAYAMA   64 ( 1 )   11 - 18   2010.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    The skeletal muscle is classified into 2 types, slow oxidative or fast glycolytic muscle. For further characterization, we investigated the capillary architecture in slow and fast muscles. The rat soleus and extensor digitorum longus (EDL) muscles were used as representatives of slow and fast muscles, respectively. To investigate capillary density, sections of both types of muscle were stained with alkaline phosphatase; the soleus muscle showed more intense reactivity, indicating that it had a denser capillary structure than the EDL muscle. We then injected fluorescent contrast medium into samples of both muscle types for light and confocal-laser microscopic evaluation. The capillary density and capillary-to-fiber ratio were significantly higher, and the course of the capillaries was more tortuous, in the soleus muscle than in the EDL muscle. Capillary coursed more tortuously in the soleus than in the EDL muscle. Succinate dehydrogenase (SDH) activity, an indicator of mitochondrial oxidative capacity, and vascular endothelial growth factor (VEGF) expression were also significantly higher in the soleus muscle. Thus, we conclude that slow oxidative muscle possess a rich capillary structure to provide demanded oxygen, and VEGF might be involved in the formation and/or maintenance of this highly capillarized architecture.

    Web of Science

    researchmap

  • A disintegrin and metalloproteinase with thrombospondin motifs 9 (ADAMTS9) expression by chondrocytes during endochondral ossification Reviewed

    Kanae Kumagishi, Keiichiro Nishida, Tomoichiro Yamaai, Ryusuke Momota, Shigeru Miyaki, Satoshi Hirohata, Ichiro Naito, Hiroshi Asahara, Yoshifumi Ninomiya, Aiji Ohtsuka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   72 ( 3 )   175 - 185   2009.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT SOC HISTOLOGY & CYTOLOGY  

    A disintegrin and metalloproteinase with thrombospondin motifs 9 (ADAMTS9) is known to influence aggrecan degradation in endochondral ossification, but its role has not been well understood. In the present study, in vitro gene expression of ADAMTS9 was investigated by RT-PCR in ATDC5 cells in which experimentally chondrogenic differentiation had been induced. We also investigated the protein localization and gene expression pattern of ADAMTS9 in the tibia growth plate cartilage of male mice in a day 1 neonate, 7-week-old young adult, and a 12-week-old adult by immunohistochemistry and in situ hybridization and compared the results with the expression of proliferating cell nuclear antigen (PCNA) and type X collagen for the identification of proliferative and hypertrophic chondrocyte phenotypes, respectively. We found the gene expression of ADAMTS9 by ATDC5 cells as a dual mode, both before the expression of type X collagen and after hypertrophic differentiation. The immunoreactivity of ADAMTS9 was observed in chondrocytes of proliferative and hypertrophic zones in the growth plate. The population of ADAMTS9 positive cells decreased with age. The results of the present study suggest that ADAMTS9 might have a role in aggrecan cleavage around the chondrocytes to allow chondrocyte proliferation and hypertrophy.

    DOI: 10.1679/aohc.72.175

    Web of Science

    PubMed

    researchmap

  • 潰瘍性大腸炎における大腸粘膜下基底膜のIV型コラーゲンα鎖の解析

    佐藤 博之, 内藤 一郎, 百田 龍輔, 小林 直哉, 大塚 愛二

    日本消化器病学会雑誌   106 ( 臨増大会 )   A827 - A827   2009.9

     More details

    Language:Japanese   Publisher:(一財)日本消化器病学会  

    researchmap

  • 肺内気道-呼吸上皮とその基底膜を構成するIV型コラーゲンのα鎖解析

    日根野谷 紀子, 内藤 一郎, 百田 龍輔, 佐渡 義一, 大塚 愛二

    解剖学雑誌   84 ( 2 )   68 - 68   2009.6

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • マウス成長軟骨におけるADAMTS-9発現とその役割について

    熊岸 加苗, 西田 圭一郎, 百田 龍輔, 山合 友一朗, 廣畑 聡, Kadir Demircan, 内藤 一郎, 二宮 善文, 大塚 愛二

    解剖学雑誌   84 ( Suppl. )   139 - 139   2009.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • 炎症性腸疾患における大腸粘膜下基底膜のIV型コラーゲンα鎖の解析

    佐藤 博之, 内藤 一郎, 斉藤 健司, 百田 龍輔, 小林 直哉, 大塚 愛二

    解剖学雑誌   84 ( Suppl. )   144 - 144   2009.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • ラット腹側尾動脈の内弾性板の加齢性変化

    品岡 玲, 百田 龍輔, 内藤 一郎, 大塚 愛二

    解剖学雑誌   84 ( Suppl. )   180 - 180   2009.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • ショウジョウバエXV/XVIII型コラーゲンDroleは正常な発生と基底膜の構造に重要である

    百田 龍輔, 内藤 一郎, 二宮 善文, 大塚 愛二

    解剖学雑誌   84 ( Suppl. )   130 - 130   2009.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • Type IV collagen alpha chains of the basement membrane in the rat bronchioalveolar transitional segment Reviewed

    Noriko Hinenoya, Ichiro Naito, Ryusuke Momota, Yoshikazu Sado, Kanae Kumagishi, Yoshifumi Ninomiya, Aiji Ohtsuka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   71 ( 3 )   185 - 194   2008.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT SOC HISTOLOGY & CYTOLOGY  

    In the present study, we have analyzed the a(IV) chain distribution in the subepithelial basement membrane (BM) of the rat pulmonary airway from the bronchi to alveoli. We have furthermore analyzed the a(IV) chain distribution in the subepithelial BM of the bronchioalveolar duct junction (BADJ) using a(IV) chain specific monoclonal antibodies. Our results show that the BM of the bronchial and bronchiolar epithelium contains [al(IV)](2) a2(IV) and [ a5(IV)](2) a6(IV) molecules and confirmed that the alveolar BM consists of [a1(IV)](2) a2(IV) and a3(IV) a4(IV) a5(IV) molecules. There are also small regions in BADJ consisting of only [ a] (IV)](2) a2(IV) molecules without a3(IV) a4(IV) a5(IV) and [ a5(IV)](2) a6(IV) molecules. Moreover, the bronchioalveolar stem cells (BASCs) -primordial cells for bronchiolar Clara cells and alveolar type II (AT2) cells- lie adjacent to such small regions. These findings suggest that [ al(IV)](2) a2(IV) may be important for the BASCs to self-renew or to self-maintain themselves and that microenvironments produced by a(IV) chains may be important for cell differentiation.

    DOI: 10.1679/aohc.71.185

    Web of Science

    PubMed

    researchmap

  • 指紋に対応する細胞外マトリックス

    内藤 一郎, 斎藤 健司, 百田 龍輔, 佐渡 義一, 二宮 善文, 大塚 愛二

    解剖学雑誌   83 ( Suppl. )   219 - 219   2008.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • ラット気管支・肺胞移行部における基底膜構成IV型コラーゲンのα鎖解析

    日根野谷 紀子, 内藤 一郎, 百田 龍輔, 佐渡 義一, 大塚 愛二

    解剖学雑誌   83 ( Suppl. )   204 - 204   2008.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • The differential distribution of type IV collagen alpha chains in the subepithelial basement membrane of the human alimentary canal Reviewed

    Hiroyuki Sato, Ichiro Naito, Ryusuke Momota, Yoshio Naomoto, Tomoki Yamatsuji, Yoshikazu Sado, Yoshifumi Ninomiya, Aiji Ohtsuka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   70 ( 5 )   313 - 323   2007.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT SOC HISTOLOGY & CYTOLOGY  

    We studied distribution patterns of type IV collagen a chains in the subepithelial basement membrane (SBM) of the human gastrointestinal tract - the esophagus through the anal canal - by immunofluorescent microscopy using alpha(IV) chain-specific monoclonal antibodies. The alpha 1(IV), alpha 2(IV), alpha 5(IV), and alpha 6(IV) chains were found in the SBM throughout the tract, indicating the localization of [alpha 1(IV)](2) alpha 2(IV) and [alpha 5(IV)](2) alpha 6(IV) heterotrimeric molecules. The [alpha 1(IV)](2) alpha 2(IV) molecule was continuously stained, while the [alpha 5(IV)]2 alpha 6(IV) molecule was weakly stained in gastric glands and small intestinal crypts. In addition, the SBM at the luminal surface epithelium of the stomach and large intestine contained small amounts of alpha 3(IV) and alpha 4(IV) chains which combined to form the alpha 3(IV) alpha 4(IV) alpha 5(IV) heterotrimeric molecule with alpha 5(IV) chain. The SBM beneath the villous epithelium of the small intestine was also demonstrated to have an alpha 3(IV) chain and alpha 4(IV) chain. Considering the specific locations of the type IV collagen trimers throughout the gastrointestinal SBM, the supramolecular network containing the alpha 3(IV) alpha 4(IV) alpha 5(IV) molecule appears to function as a selective permeability barrier and/or as a protection against chemical stress from the luminal digestive enzymes.

    DOI: 10.1679/aohc.70.313

    Web of Science

    PubMed

    researchmap

  • Intermediate filaments of endoskeleton within human erythrocytes. Reviewed

    Kazutaka Terasawa, Takehito Taguchi, Ryusuke Momota, Ichiro Naito, Aiji Ohtsuka

    BLOOD   110 ( 11 )   516A - 516A   2007.11

     More details

    Language:English   Publisher:AMER SOC HEMATOLOGY  

    DOI: 10.1182/blood.V110.11.1734.1734

    Web of Science

    researchmap

  • The distribution of type IV collagen alpha chains in the mouse ovary and its correlation with follicular development Reviewed

    Kazuyo Nakano, Ichiro Naito, Ryusuke Momota, Yoshikazu Sado, Haruko Hasegawa, Yoshifumi Ninomiya, Aiji Ohtsuka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   70 ( 4 )   243 - 253   2007.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT SOC HISTOLOGY & CYTOLOGY  

    The present study aims to identify a chains of type IV collagen in the basement membrane of the mouse ovarian follicle and examine their changes during follicular development using immunofluorescence microscopy with specific monoclonal antibodies. The basement membrane of the serous mesothelium enveloping the ovary contained all a chains of type IV collagen, alpha 1(IV) through alpha 6(IV) chains. Primordial follicles showed a distinct immunoreactivity against all six a chains in their basement membranes. Immunolabeling for alpha 3(IV) and alpha 4(IV) chains was almost eliminated in the primary follicles. In basement membranes of secondary and Graafian follicles, the immunofluorescent reaction of a3(IV) and a4(IV) chains disappeared in Graafian follicles, a partial reduction in fluorescent immunostaining intensity to alpha 5(IV) and alpha 6(IV) chains was observed; only alpha 1(IV) and alpha 2(IV) chains were not degraded throughout follicular development. On atretic follicles, in addition to alpha 1(IV) and a2(IV) chains, alpha 3(IV), alpha 4(IV), alpha 5(IV) and alpha 6(IV) chains frequently persisted. A basement membrane-like matrix within the follicular granulosa cell layer, such as the focimatrix (focal intraepithelial matrix) and/or Call-Exner body, was also recognized in mouse secondary and Graafian follicles and contained alpha 1(IV), alpha 2(IV), alpha 5(IV) and alpha 6(IV) chains but not alpha 3(IV) and alpha 4(IV) chains. We expect that the decrease in alpha(IV) chains prompts follicular development and is a prerequisite condition for follicular maturation.

    DOI: 10.1679/aohc.70.243

    Web of Science

    PubMed

    researchmap

  • The distributions of type IV collagen alpha chains in basement membranes of human epidermis and skin appendages Reviewed

    Haruko Hasegawa, Ichiro Naito, Kazuyo Nakano, Ryusuke Momota, Keiichiro Nishida, Takehito Taguchi, Yoshikazu Sado, Yoshifumi Ninomiya, Aiji Ohtsuka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   70 ( 4 )   255 - 265   2007.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT SOC HISTOLOGY & CYTOLOGY  

    Distributions of type IV collagen a chains in the basement membrane (BM) of human skin and its appendages were analyzed by immunofluorescent microscopy using chain-specific monoclonal antibodies. The basement membrane beneath the epidermis contained [alpha 1(IV)]2 alpha 2(IV) and [ alpha 5(IV)]2 alpha 6(IV) but no alpha 3(IV) alpha 4(IV) alpha 5(IV); this held true for at the eccrine sweat glands and glandular ducts, sebaceous glands, hair follicles, and arrector muscles of hair. The secretary portion of the eccrine sweat glands was rich in [alpha 1(IV)]2 alpha 2(IV) and had less [ alpha 5(IV)]2 alpha 6(IV), while [alpha 5(IV)]2 alpha 6(IV) was abundant in the ductal portion. In the subepidermal zone, alpha 5(IV)/ alpha 6(IV) chain negative spots (1.9-15.0 mu m) were frequently observed. Triple staining samples (Mel.2, alpha 2(IV) and alpha 5(IV) chains) showed that about 50% of epidermal melanocytes colocalized with such spots. Results suggest that these alpha 5(IV)/ alpha 6(IV) chain negative spots of the subepidermal basement membrane have a particular relationship with melanocytes and are sites for certain interactions between the two.

    DOI: 10.1679/aohc.70.255

    Web of Science

    PubMed

    researchmap

  • 赤血球の両凹形態を維持する細胞質内骨格(Cytoplasmic endoskeleton maintaining the biconcave shape of erythrocytes)

    寺沢 和貴, 田口 勇仁, 百田 龍輔, 内藤 一郎, 大塚 愛二

    解剖学雑誌   82 ( 3 )   114 - 114   2007.9

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • ブタの神経筋接合部に局在するIV型コラーゲンα鎖の解析

    山内 健太朗, 内藤 一郎, 森 雄市, 百田 龍輔, 長谷川 治子, 二宮 善文, 佐渡 義一, 田口 勇仁, 野村 信介, 大塚 愛二

    解剖学雑誌   82 ( 3 )   112 - 112   2007.9

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • ショウジョウバエXV/XVIII型コラーゲンの生物学的機能

    百田 龍輔, 内藤 一郎, 二宮 善文, 大塚 愛二

    日本結合組織学会学術大会・マトリックス研究会大会合同学術集会プログラム・抄録集   39回・54回   78 - 78   2007.5

     More details

    Language:Japanese   Publisher:日本結合組織学会・マトリックス研究会  

    researchmap

  • マウス成長軟骨におけるADAMTS-9発現の検討

    熊岸 加苗, 西田 圭一郎, 百田 龍輔, 山合 友一朗, 広畑 聡, Demircan Kadir, 内藤 一郎, 二宮 善文, 大塚 愛二

    解剖学雑誌   82 ( Suppl. )   150 - 150   2007.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • IV型コラーゲンα鎖特異抗体反応基の抗原賦活法

    森 雄一, 内藤 一郎, 百田 龍輔, 二宮 善文, 大塚 愛二

    解剖学雑誌   82 ( Suppl. )   271 - 271   2007.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • ヒト赤血球には内部細胞骨格が見られ、βアクチンとニューロフィラメント蛋白を含む

    寺沢 和貴, 田口 勇仁, 百田 龍輔, 内藤 一郎, 大塚 愛二

    解剖学雑誌   82 ( Suppl. )   231 - 231   2007.3

     More details

    Language:English   Publisher:(一社)日本解剖学会  

    researchmap

  • ラットの腹側尾動脈の血管鋳型標本に見られた風変わりな刻印

    品岡 玲, 百田 龍輔, 大塚 愛二, 内藤 一郎

    解剖学雑誌   82 ( Suppl. )   150 - 150   2007.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • Human erythrocytes possess a cytoplasmic endoskeleton containing beta-actin and neurofilament protein Reviewed

    Kazutaka Terasawa, Takehito Taguchi, Ryusuke Momota, Ichiro Naito, Takuro Murakami, Aiji Ohtsuka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   69 ( 5 )   329 - 340   2006.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT SOC HISTOLOGY & CYTOLOGY  

    The biconcave disc shape of mammalian erythrocytes has been considered to be maintained only with a membrane underlain by a membranous cytoskeleton. Our improved ion-etching/scanning electron microscopy and saponin-ethanol treatment combined with immunocytochemistry in the human red blood cell revealed the three-dimensional structure of this cytoplasmic endoskeleton apart from the classical membranous cytoskeleton. The endoskeletal meshwork images obtained by the saponin-ethanol treatment corresponded to those by the repeated ion-etching method. The actin-rich endoskeleton was divided into two layers, one superficial and the other deep. The superficial filaments were perpendicularly connected to the membranous cytoskeleton, while the deep filaments formed an irregularly directed complicated meshwork. In the transitional hillside region between the convex periphery and concave center, the endoskeletal filaments containing a neurofilament protein ran parallel to the hillside slope toward the concave center. The endoskeleton of the erythrocyte associating with the membranous cytoskeleton may serve to keep its unique biconcave disc shape deformable, pliable, and restorable against external circumstances.

    DOI: 10.1679/aohc.69.329

    Web of Science

    researchmap

  • ADAMTS-ECM interaction modulates BMP developmental control Reviewed

    J. Fessler, R. Momota, C. Cresse, K. Chavan, L. Fessler

    MATRIX BIOLOGY   25   S30 - S30   2006.11

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.matbio.2006.08.084

    Web of Science

    researchmap

  • 血管基底膜IV型コラーゲンの免疫組織学的解析 共焦点レーザー顕微鏡を用いた三次元観察

    森本 尊雅, 内藤 一郎, 百田 龍輔, 芳原 成美, 長谷川 治子, 田口 勇仁, 西田 圭一郎, 佐渡 義一, 二宮 善文, 大塚 愛二

    解剖学雑誌   81 ( Suppl. )   155 - 155   2006.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • 基底膜の多様性 α(IV)鎖特異抗体をもちいた基底膜IV型コラーゲンα鎖の多様性解析

    内藤 一郎, 長谷川 治子, 森本 尊雅, 百田 龍輔, 斉藤 健司, 佐渡 義一, 二宮 善文, 大塚 愛二

    解剖学雑誌   81 ( Suppl. )   110 - 110   2006.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • In situ preparation of colloidal iron by microwave irradiation for transmission electron microscopy Reviewed

    S Nakatani, Naito, I, R Momota, N Hinenoya, K Horiuchi, K Nishida, A Ohtsuka

    ACTA MEDICA OKAYAMA   60 ( 1 )   59 - 64   2006.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    We attempted to prepare colloidal iron within tissues by means of microwave irradiation. Mouse tissue blocks were fixed with a mixture of paraformaldehyde and ferric chloride in a cacodylate buffer, immersed in a cacodylate buffered ferric chloride solution, and irradiated in a microwave processor. Colloidal iron was prepared within tissues or cells, and was observed in the form of electron dense fine granules (1-2 nm in diameter) by transmission electron microscopy. Collagen fibrils in the connective tissue showed colloidal iron deposition at regular periodical intervals. Cells in the splenic tissue showed that fine colloidal granules were deposited on the ribosomes but not on the nuclear chromatin. This finding suggests that ferric ions could not diffuse into the nucleus, which was surrounded by the nuclear envelope. The podocyte processes of the renal glomerulus were stained diffusedly. Though this microwave in situ colloidal iron preparation method has some limitations, it is convenient for use in biomedical specimen preparation in transmission electron microscopy.

    DOI: 10.18926/AMO/30753

    Web of Science

    PubMed

    researchmap

  • In situ preparation of colloidal iron by microwave irradiation for transmission electron microscopy Reviewed

    S Nakatani, Naito, I, R Momota, N Hinenoya, K Horiuchi, K Nishida, A Ohtsuka

    ACTA MEDICA OKAYAMA   60 ( 1 )   59 - 64   2006.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    We attempted to prepare colloidal iron within tissues by means of microwave irradiation. Mouse tissue blocks were fixed with a mixture of paraformaldehyde and ferric chloride in a cacodylate buffer, immersed in a cacodylate buffered ferric chloride solution, and irradiated in a microwave processor. Colloidal iron was prepared within tissues or cells, and was observed in the form of electron dense fine granules (1-2 nm in diameter) by transmission electron microscopy. Collagen fibrils in the connective tissue showed colloidal iron deposition at regular periodical intervals. Cells in the splenic tissue showed that fine colloidal granules were deposited on the ribosomes but not on the nuclear chromatin. This finding suggests that ferric ions could not diffuse into the nucleus, which was surrounded by the nuclear envelope. The podocyte processes of the renal glomerulus were stained diffusedly. Though this microwave in situ colloidal iron preparation method has some limitations, it is convenient for use in biomedical specimen preparation in transmission electron microscopy.

    Web of Science

    researchmap

  • Differential expression of mouse alpha 5(IV) and alpha 6(IV) collagen genes in epithelial basement membranes Reviewed

    K Saito, Naito, I, T Seki, T Oohashi, E Kimura, R Momota, Y Kishiro, Y Sado, H Yoshioka, Y Ninomiya

    JOURNAL OF BIOCHEMISTRY   128 ( 3 )   427 - 434   2000.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    We first completed the primary structure of the mouse alpha 5(IV) and alpha 6(IV) chains, from which synthetic peptides were produced and alpha chain-specific monoclonal antibodies were raised. Expression of collagen IV genes in various basement membranes underlying specific organ epithelia was analyzed by immunohistochemical staining using these monoclonal antibodies and other antibodies from human and bovine sequences. It was possible to predict the presence of the three collagen TV molecules: [alpha 1(IV)](2) alpha 2(IV), alpha 3(IV)alpha 4(IV)alpha 5(IV), and [alpha 5(IV)](2)alpha 6(IV), In skin basement membrane two of the three forms, [alpha 1(IV)](2)alpha 2(IV) and [alpha 5(IV)](2)alpha 6(IV), were detected. The alpha 3(IV)alpha 4(IV)alpha 5(IV) molecule was observed as the major form in glomerulus, alveolus, and choroid plexus, where basement membranes function as filtering units. The molecular form [alpha 5(IV)](2)alpha 6(IV) was present in basement membranes in tubular organs such as the epididymis, where the tubes need to expand in diameter. Thus, the distribution of the basement membranes with different molecular composition is consistent with tissue-specific function.

    Web of Science

    researchmap

  • Organization and expression of basement membrane collagen IV genes and their roles in human disorders Reviewed

    Y Sado, M Kagawa, Naito, I, Y Ueki, T Seki, R Momota, T Oohashi, Y Ninomiya

    JOURNAL OF BIOCHEMISTRY   123 ( 5 )   767 - 776   1998.5

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    Six distinct genes have been identified as belonging to the type IV collagen gene family. They can be organized into three sets, i.e., COL4A1/COL4A2, COL4A3/COL4A4, and COL4A5/COL4A6, which are localized on three different chromosomes in humans, 13, 2, and X, respectively, Within each set the genes are aligned head-to-head and their expression is regulated by bidirectional promoters between the genes. Transcriptional regulation of the COL4A1/COL4A2 set has been well characterized, The transcription of COL4A6 seems to be controlled by two alternative promoters. While collagen IV molecules composed of alpha 1 and alpha 2 chains are broadly distributed, molecules comprising combinations of the other four chains, alpha 3-alpha 6, are important components of specialized basement membranes, The precise chain composition of triple-helical molecules assembled from the alpha 3-alpha 6 chains is not entirely clear, but it is hypothesized that alpha 3-alpha 5 chains and alpha 5 and alpha 6 chains form heterotrimeric molecules. Several pieces of evidence indicate that alpha 3/alpha 4/alpha 5 molecules and alpha 5/alpha 6 molecules are components of the basement membrane network, This helps explain the observation that the kidney and skin basement membranes from patients with Alport syndrome caused by mutations in the alpha 5 coding gene, COL4A5, are defective in the alpha 3, alpha 4, and alpha 6 chains together with the alpha 5 chain. Large deletions involving the COL4A5 and COL4A6 genes have been found in rare cases of diffuse leiomyomatosis associated with Alport syndrome.

    Web of Science

    researchmap

  • Two genes, COL4A3 and COL4A4 coding for the human alpha 3(IV) and alpha 4(IV) collagen chains are arranged head-to-head on chromosome 2q36 Reviewed

    R Momota, M Sugimoto, T Oohashi, K Kigasawa, H Yoshioka, Y Ninomiya

    FEBS LETTERS   424 ( 1-2 )   11 - 16   1998.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    We first isolated and characterized genomic DNA fragments that cover the 5' flanking sequences of COL4A3 and COL4A4 encoding the human basement membrane alpha 3(IV) and alpha 4(IV) collagen chains, respectively. Nucleotide sequence analysis indicated that the two genes are arranged head-to-head. To determine transcription start site for COL4A4 gene, we performed RACE and RNase protection assays, indicating that there are two alternative transcripts presumably derived from two different promoters. Interestingly, one transcription start site (from exon 1') of COL4A4 is only 5 bp away from the reported transcription start site of COL4A3, whereas the other transcript (from exon 1) starts 373 nucleotides downstream from the first one, generating the two kinds of transcripts that differ in the 5' UTR regions. Expression of these two transcripts appears tissue-specific; exon 1 transcript was expressed predominantly in epithelial cells, while exon 1' transcript showed rather ubiquitous and low expression. The nucleotide sequence of the promoter region is composed of dense CpG dinucleotides, GC boxes, CTC boxes and a CCAAT box but no TATA box. These results provide information to delineate the promoter activity for the tissue-specific expression of the six type IV collagen genes and basement membrane assembly in different tissues and organs. (C) 1998 Federation of European Biochemical Societies.

    DOI: 10.1016/S0014-5793(98)00128-8

    Web of Science

    researchmap

▼display all

Books

  • 漢字で覚える解剖ドリル (プチナースBOOKS)

    菊地よしこ, 百田, 龍輔( Role: Joint author)

    照林社  2017.4  ( ISBN:4796523995

     More details

    Total pages:104   Language:Japanese

    CiNii Books

    ASIN

    researchmap

MISC

  • Possible metabolic changes in the liver of Collagen XVIII knock out mice

    百田龍輔, 米澤朋子, 柳井広之, 大橋俊孝

    日本結合組織学会学術大会抄録集   54th (Web)   2022

  • 解剖研究を通じた超音波による長母指屈筋副頭の検出方法の改善

    坂口和輝, 坂口和輝, 中原龍一, 百田龍輔, 小阪美津子, 品岡玲, 大塚愛二

    日本解剖学会総会・全国学術集会講演プログラム・抄録集   125th   2020

  • Different effects of aerobic and anaerobic exercise on the capillary in the extensor digitorum longus: PGC-1a, VEGF, Flk-1 and Capillary Architecture

    Shinichiro Murakami, Naoto Fujita, Takeshi Morifuji, Miho Kanasasi, Masahiro Sakita, Hiroyo Kondo, Ryusuke Momota, Aiji Ohtsuka, Hidemi Fujino

    FASEB JOURNAL   27   2013.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:FEDERATION AMER SOC EXP BIOL  

    Web of Science

    researchmap

  • 微小血管内皮下弾性線維構造とその血行動態との関係

    品岡玲, 百田龍輔, 小阪美津子, 内藤一郎, 中原龍一, 大塚愛二

    日本結合組織学会学術大会抄録集   43rd   2011

  • マウス子宮基底膜を構成するIV型コラーゲンα鎖の免疫組織学的解析

    内藤 一郎, 大貫 秀策, 中橋 いずみ, 斎藤 健司, 稲垣 純子, 二宮 義文, 百田 龍輔, 大塚 愛二, 中塚 幹也

    解剖学雑誌   85 ( 2 )   83 - 83   2010.6

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • Abnormality of capillarity and oxidative enzyme activity in slow muscle of type 2 diabetic rats

    Shinichiro Murakami, Hidemi Fujino, Hiroyo Kondo, Isao Takeda, Ryusuke Momota, Kanae Kumagishi, Aiji Ohtsuka

    FASEB JOURNAL   24   2010.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:FEDERATION AMER SOC EXP BIOL  

    Web of Science

    researchmap

  • 性成熟に伴う子宮内膜基底膜IV型コラーゲンα鎖の変化

    内藤 一郎, 中橋 いずみ, 大貫 秀策, 安藤 さやか, 渡邉 美里, 斎藤 健司, 稲垣 純子, 百田 龍輔, 中塚 幹也, 二宮 善文, 大塚 愛二

    解剖学雑誌   85 ( Suppl. )   115 - 115   2010.3

     More details

    Language:Japanese   Publisher:(一社)日本解剖学会  

    researchmap

  • 基底膜のIV型コラーゲンα鎖の消化管粘膜上皮における多様性

    佐藤博之, 内藤一郎, 百田龍輔, 大塚愛二

    解剖学雑誌   83 ( 2 )   61 - 61   2008.6

     More details

  • Expression of col4a5 and col4a6 genes in basement membranes in mouse testis.

    SAITO Kenji, SEKI Tsugio, OOHASHI Toshitaka, MOMOTA Ryusuke, NINOMIYA Yoshifumi

    21   430 - 430   1998.12

     More details

  • COL4A3 and COL4A4 are arranged head-to-head and COL4A4 has two alternative promoters.

    Momota Ryusuke, Oohashi Toshitaka, Ninomiya Yoshifumi

    Connective tissue   30 ( 2 )   135 - 135   1998.6

     More details

    Language:Japanese   Publisher:The Japanese Society for Connective Tissue  

    CiNii Article

    CiNii Books

    researchmap

  • Alternative transcripts are also found in the human COL4A4 gene expression.

    R Momota, T Oohashi, M Sugimito, Naito, I, Y Sado, Y Ninomiya

    MATRIX BIOLOGY   15 ( 3 )   171 - 172   1996.9

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:GUSTAV FISCHER VERLAG  

    Web of Science

    researchmap

▼display all

Presentations

  • 人体解剖学デジタル教材の開発と応用 Invited

    大塚愛二、百田龍輔

    第126回 日本解剖学会総会  2021.3 

     More details

    Presentation type:Oral presentation (invited, special)  

    researchmap

  • Ocular abnormalities and the change of gene expression in Alport syndrome model mice

    Tang Shaoying, Tomoko Yonezawa, Ryusuke Momota, Hiroshi Matsumae, Yuki Kanzaki, Yuki Morizane, Toshitaka Oohashi

    2020.9 

     More details

    Presentation type:Oral presentation (general)  

    researchmap

  • 人体解剖学デジタル教材の開発と応用 Invited

    大塚愛二, 百田龍輔

    第125回 日本解剖学会総会  2020.3 

     More details

    Presentation type:Oral presentation (invited, special)  

    researchmap

  • Transcriptional changes of the skin during aging process

    Ryusuke Momota, Tomoko Yonezawa, Toshitaka Oohashi, Zenzo Isogai

    2019.6 

     More details

    Presentation type:Poster presentation  

    researchmap

  • Developmental studies of Drole, Drosophila type XV/XVIII collagen. Invited

    Ryusuke Momota, Ichiro Naito, Yoshifumi Ninomiya, Aiji Ohtsuka

    8th Pan-Pacific Connective Tissue Societies Symposium  2009.7 

     More details

    Language:English   Presentation type:Oral presentation (invited, special)  

    researchmap

  • ADAMTS_ECM interaction modulates BMP developmental control.

    John H. Fessler, Ryusuke Momota, Chino-Kumagai Cresse, Kavita Chavan, Liselotte I. Fessler

    The 2006 ASMB Biennial meeting  2006.10 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    researchmap

  • New ECM-metalloprotease interactions during development

    Ryusuke Momota, Liselotte I. Fessler, Irina Kramerova, John H. Fessler

    6th Pan-Pacific Connective Tissue Societies Symposium, Hawaii, USA  2005.10 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    researchmap

▼display all

Research Projects

  • 1細胞解析による基底膜の形成機序を軸とした再上皮化メカニズムの解明

    Grant number:23K09100  2023.04 - 2026.03

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    米澤 朋子, 大野 充昭, 百田 龍輔

      More details

    Grant amount:\4680000 ( Direct expense: \3600000 、 Indirect expense:\1080000 )

    researchmap

  • Functional deteriorations of liver/pancreas caused by age-related changes of type XVIII collagen

    Grant number:20K11556  2020.04 - 2023.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    百田 龍輔, 米澤 朋子, 大塚 愛二, 大橋 俊孝, 内藤 一郎

      More details

    Grant amount:\4290000 ( Direct expense: \3300000 、 Indirect expense:\990000 )

    脂肪肝の症状がはっきりとみられるXVIII型コラーゲン欠損老齢マウスだけでなく、若い欠損マウスで”予兆”のような現象を改善することで脂肪肝、NAFLDへの進行を予防できるのではないかと考え、加齢マウスだけでなく2ヶ月齢の若いマウスを用いて改めて、網羅的に遺伝子発現と組織学的な検討を行った。若い欠損マウスは細胞内脂肪滴の蓄積では組織学的に大きな差は認められなかったが、血管周囲の構造について野生型と比較して若干違いが見られたのでその再現性について確認を行っている。
    さらに、野生型、欠損の若いマウスについても、双方の肝臓からRNAを抽出し、RNA-seqにより全転写産物の解析を進めている。前年度の加齢マウスで行ったネットワーク解析、コンピューターによる代謝シミュレーションを行い、代謝経路・産物について経時的にどう変化するかについて検討する予定である。
    また、新たにリピドミクスの系を立ち上げたので、マウスの血液中や肝臓に蓄積する脂質組成について比較検討を行っている。すなわち、上記のコンピューターシミュレーションの結果を生体で実際に起きているかどうか確認を進めている。
    一方、上記のシミュレーション結果をもとに、ネットワーク解析などから中心的な役割を果たす複数の代謝経路、遺伝子産物、代謝産物を同定した。これらの候補遺伝子産物で薬理学的な調節が可能な候補を抽出した。また、その機能を調節できるような化合物検索をバーチャルに行う高性能GPU搭載コンピューターの解析環境を立ち上げた。現在これらの結果をもとに論文作成に取り掛かっている。

    researchmap

  • 皮膚創傷治癒における基底膜コラーゲンの機能解析と治療応用のための基盤的研究

    Grant number:20K09867  2020.04 - 2023.03

    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)

    米澤 朋子, 百田 龍輔, 稲川 喜一

      More details

    Grant amount:\4290000 ( Direct expense: \3300000 、 Indirect expense:\990000 )

    本研究は創傷後の表皮形成に重要な細胞外マトリックスの役割を調べ、治癒を促す分子メカニズムを明らかにすることを目的としている。細胞外マトリックスを構成する成分は質的にも量的にも創傷の治癒過程で変化することが知られており、その変化は治癒に有効あるいは不利な細胞外環境であると考えられ、細胞の応答を制御すると考えられる。我々は創傷部の周囲にスプリントを取り付けることによって、治癒の過程を再現性良く詳細な解析が可能となるマウス創傷治癒モデルを作製した。モデルの確立には過去の論文を参考にしたが、いくつかの改良を加えている。このマウス皮膚創傷治癒モデルを用いた実験から、再生する表皮の先端部にかなり早期に出現する細胞外マトリックス成分に着目した。早期の発現が認められたXVIII型コラーゲンは、これまでの様々な研究から特徴的な分子構造を持ち多機能性が示唆されているが、皮膚創傷治癒での役割については十分に解明されていない。2021年度は遺伝子改変マウスと野生型マウスを用いて創傷治癒モデルを作製し、組織学的な解析と免疫組織化学染色や定量的PCRを行い、再生上皮形成と創収縮におけるXVIII型コラーゲンの役割について解析を行った。透過型電子顕微鏡による観察も加えて行った。さらに、マウスから表皮角化細胞を初代培養し、細胞接着や移動能における機能解析を行うとともに、網羅的な遺伝子発現解析に着手した。また、リコンビナントタンパク質の作製を行うためにXVIII型コラーゲン安定発現株を確立した。リコンビナントタンパク質を用いた機能の解析を行う予定である。

    researchmap

  • 人工知能による深層学習を応用した運動障害性咀嚼障害の多軸診断支援システムの開発

    Grant number:20K10071  2020.04 - 2023.03

    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)

    大野 彩, 窪木 拓男, 森田 瑞樹, 菊谷 武, 百田 龍輔

      More details

    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    本研究では、①咀嚼運動をセンサー付きカメラで撮影した画像から評価できる「運動性咀嚼機能評価プロトコール」を開発し,信頼性・妥当性を確認すること,②開発した「運動性咀嚼機能評価プロトコール」を用いて,咀嚼障害患者および正常咀嚼者の撮影および診断を行い,教師データを収集すること,そして,③教師データを人工知能学習に供し,人工知能による運動障害性咀嚼障害診断システムを開発することを目的としている.摂食時に撮影した画像から咀嚼・嚥下運動の評価を行うために,本年度は開発を進めてきた撮影用アプリケーションを利用し,撮影を開始した.そして撮影結果を用いて,運動評価の精度検証および人工知能学習を開始している.
    これまでに撮影した画像から運動データの精度検証を行い,撮影角度や撮影時間の最適化を行った.そして,それらの情報を元に撮影用アプリケーションの改修を行った.また,撮影時に即時データ収集する最適座標を絞り,即時データ収集用サーバーの構築を行った.そして,主に外来に通院可能な健常高齢者を対象とし,開発した撮影用アプリケーションを利用したデータ収集を開始している.また,身長,体重,体組成データ,全身疾患,身体・認知機能,栄養状態,食形態,摂取食品,摂取カロリー,義歯の使用状況,グミゼリー咀嚼機能検査,臼歯部移送試験,舌圧測定,オーラルディアドコキネシス,生活環境,家族構成,食事時間等のデータ収集を行っている.

    researchmap

  • The functions of collagen XVIII in skin aging

    Grant number:17K01848  2017.04 - 2020.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Momota Ryusuke

      More details

    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    We analyzed transcriptomic data of human skin to find transcripts whose expression levels showed significant changes in age-dependent manner. These transcripts involve in biological functions such as RNA transcription, splicing, transmembrane, lipoproteins, mitochondrial components, epithelial keratinization, and microtubules. We found that fresh unripe peach extract significantly improved mRNA levels and partially localizations of basement membrane components.
    We had established a very simple less invasive method to detect collagen types XVIII and IV. In addition, we had established a method to detect immature epidermal cells which appear during barrier formation after skin damage and wrote a computational script for quantification.

    researchmap

  • Analysis of trans-plasma membrane molecular machinery between collagen types XV/XVIII and mitochondria

    Grant number:26350892  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Momota Ryusuke, OOHASHI Toshitaka, YONEZAWA Tomoko, KOMIYAMA Takaaki, NARASAKI Masahiro

      More details

    Grant amount:\4810000 ( Direct expense: \3700000 、 Indirect expense:\1110000 )

    To identify the molecules which would compose "the tran-plasma membrane molecular machinery" of collagen types XV/XVIII and mitochondria, we have screened thousands of genes expressed in muscular tissues and identified possible candidate G-protein coupled receptors. In addition, our histological analyses on collagen types XV/XVIII mutant animals revealed abnormal structures of cytoskeletons, which would potentially compromise the mitochondrial and cellular functions. Moreover, we had successfully optimized the immunohistochemical conditions for the anti-collagen XV/XVIII monoclonal antibodies, which enabled us to use variously processed tissue samples to diagnose human diseases caused by defective collagen XV/XVIII.

    researchmap

  • Role of basement membranes in small-vessel disease of brain

    Grant number:23390348  2011.04 - 2014.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    NINOMIYA YOSHIFUMI, YONEZAWA Tomoko, OOHASHI Toshitaka, HIROHATA Satoshi, OHTSUKA Aiji, HATANAKA Kunihiko, SAITO Kenji, MOMOTA Ryusuke, OGAWA Hiroko, INAGAKI Jyunko

      More details

    Grant amount:\19240000 ( Direct expense: \14800000 、 Indirect expense:\4440000 )

    Abnormalities of the basement membranes in brain small vessels are considered to cause the break-down of blood-brain barrier (BBB) and intracerebral hemorrhage. We established the mouse encephalopathy model by the administration of TNF-alpha intravenously, in which BBB permeability increase transiently. To know the change of type IV collagen, as major component of the basement membranes, in the encephalopathy model, we performed Western blot and immunohistochemistry. The results suggested that the degradation of type IV collagen was associated with the break-down of BBB.

    researchmap

  • 聴覚におけるペリニューロナルネットの役割ー聴覚伝導路特異的Bral2の機能解析ー

    Grant number:22591879  2010 - 2012

    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)

    別宮 洋子, 大橋 俊孝, 大塚 愛二, 百田 龍輔

      More details

    Grant amount:\3640000 ( Direct expense: \2800000 、 Indirect expense:\840000 )

    Bral2は、リンクプロテイン(LP)の一種で、ヒアルロン酸(HA)結合型コンドロイチン硫酸プロテオグリカン(CSPG)およびHAとともに複合体を形成する。LPは、その複合体形成及び安定化に必須の分子であると考えられている。Bral2は、CSPGの中でもブレビカンと共局在する事がわかっている。これらは、成体脳ではペリニューロナルネット(PNN)と呼ばれる、神経細胞周囲の網目状構造に存在する。
    本研究では、聴覚伝導路において、Bral2複合体のシナプス伝達への関わりを1)シナプスの固定、2)イオンプールとしての可能性、3)イオンチャネルとの分子間相互作用という観点から解析し、聴覚伝導のメカニズムの一端を解明する事を目的とした。
    (I)Bral2欠損マウスにおける神経細胞体への影響(大塚・別宮)
    Bral2欠損マウスにおいて、PNNを構成するCSPG複合体構成分子が小脳核では局在できないことがわかっている。この神経核においてシナプスの接着等に何らかの形態的変化がないかを、電子顕微鏡での観察によって詳細に調べた。その結果、Bral2の小脳核において、単位面積当たりにおけるシナプス数が有意に減少していることが確認された。
    (II)Bral2タンパクの発現機構(百田・大橋)
    Bral2は、in situ hybridization等の結果から、mRNA発現神経細胞が投射した先の神経周囲にタンパクを産出していることが示唆されている。この発現機構を調べるために、Bral2のリコンビナントタンパクを神経細胞に発現させ、軸索輸送により投射先にタンパクが発現されるのかを解析したところ、確かに軸策輸送が観察された。軸索輸送に関与していると予測された配列を欠くリコンビナントタンパクも発現させたが、その配列によるものではないという結果が得られた。
    (III)Bral2欠損マウスにおけるCSPG複合体構成分子への影響(別宮)
    Bral2欠損マウスにおいて、聴覚伝導路におけるCSPG複合体構成分子が、各神経核または神経のサブタイプで異なる変化を示すという結果を得た。ブレビカンは、Bral2欠損マウスにおいてPNNパターンを維持できないことから、その局在はBral2に依存していることがわかった。これらCSPGの構成に関して、解析が単純であるランビエ絞輪においてその不均一性を更に解析し、その結果を論文にまとめた。現在印刷中である。

    researchmap

  • Age-related rupture of internal elastic lamina in atherosclerosis: ultrastrucutral analysis of its occurrence and healing.

    Grant number:22591542  2010 - 2012

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    OHTSUKA Aiji, TAGUCHI Takehito, MOMOTA Ryusuke

      More details

    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    At first, we newly developed scanning electron microscopy technique using a formic acid digestion with vascular casts, which revealed, in detail, the three-dimensional(3D) architecture of delicate elastic fibers in small vessels. This method could successfully visualized the three-dimensional architecture of elastic fibers in small various sized of blood vessels, even arterioles and venules. Furthermore, the new method revealed the 3D architectures of corrupted internal elastic lamina.

    researchmap

  • ADAMTSによるアミロイド前駆体蛋白のプロセッシングと細胞外基質への影響

    Grant number:13877097  2001 - 2002

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    廣畑 聡, 米澤 朋子, 大橋 俊孝, 二宮 善文, 百田 龍輔

      More details

    Grant amount:\2000000 ( Direct expense: \2000000 )

    ADAMファミリーのうち、ADM-10とADAM-17はαセクレターゼ機能を持つことが知られているが、ADAMTSが同様な機能を持つか検討した。
    まず、基礎的実験としてADAMTS-1はその遺伝子発現がリポポリサッカライド刺激により、各臓器において発現が上昇することから炎症において何らかの役割を持つことが示唆されている。そこで、ラット実験的心筋梗塞モデルを用いてADAMTS-1の発現形式をノーザンブロット法にて検討した。ADAMTS-1は非梗塞心臓では弱い発現しか認めなかったが、梗塞心においては、梗塞後6時間でその発現が大きく上昇していた。
    マウス脳におけるADAMTSの発現をADAMTS-1〜7において検討したが、いずれもそれほど強くなかった。海外共同研究として、アミロイド前駆体蛋白を強制発現し、恒常的に発現する細胞株を樹立している、アラバマ大学Fukuchi教授らと共同実験を開始した。同細胞株は、通常の神経系細胞よりも過剰にアミロイド前駆体蛋白を発現している。これまでの検討によって過剰なアミロイド蛋白前駆体が細胞表面及び培養上清中に存在することが確認された。この実験系において過剰なアミロイド前駆体の切断がαセクレターゼによって制御されているかどうかを検討する目的でこの細胞系におけるαセクレターゼ発現の検討を開始した。実験の条件検討が複雑であり、切断の確認は困難であった。今後は、In vivoの系における実験系の確立およびアルツハイマー脳におけるADAMTSの発現解析が重要と考えられた。

    researchmap

  • Identifying new aggrecanase and producing antibody and gene-targeting mouse

    Grant number:13470312  2001 - 2002

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    HIROHATA Satoshi, YONEZAWA Tomoko, OOHASHI Toshitaka, NINOMIYA Yoshufumi, MOMOTA Ryusuke

      More details

    Grant amount:\14000000 ( Direct expense: \14000000 )

    To investigate new aggrecanase, we have done the following experiments and got some data.
    1) Various ADAMTS-specific primers were designed and RT-PCR was performed. We cultured human chondrosarcoma cell lines OUMS-27 (Okayama University Medical School-27) and stimulated with interleukine-1 beta (IL-1β). To determine gene expression quantitatively, we employed real-time RT-PCR method. Briefly, total RNA was extracted and then DNAse treatmeat was done to eliminate contaminating genomic DNA. After reverse transcribed with random primers and enzymes, cDNA was served as a template for RT-PCR. GAPDH was used for the internal control.
    2) The expression and gene regulation by IL-1β was different amonf the ADAMTS-1,4, and -5, which were reported to have aggrecan cleaving property in vitro. We also investigated other ADAMTS gene expressions.
    3) We identified another up-regulating ADAMTS gene by IL-1β in OUMS-27 cells. We raised polyclonal antibody against this new ADAMTS gene using peptide sequence of this ADAMTS.
    4) We also started to making knock-out mouse for this gene. We screened mouse genomic library and identified several clones including this new ADAMTS gene. Under the collaboration with Dr. Apte's lab in the USA, we started to put our clones to ES cells.

    researchmap

  • Functional Analysis of Basement Membrane Collagen Genes

    Grant number:11694280  1999 - 2001

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    NINOMIYA Yoshifumi, YONEZAWA Tomoko, MOMOTA Ryusuke, OOHASHI Toshitaka

      More details

    Grant amount:\8200000 ( Direct expense: \8200000 )

    By analyzing Col4a knockout mice, it is now possible to predict the biological function of α(IV) chains in vivo. We have now knockout mice of two strains: col4a3 and col4a6. Since we have evidence that there are major three molecular forms in basement membranes: (α1)2α2, α3α4α5, and (α5)2α6, it is now possible to set-up experiments to presume in vivo function of several α chains. We also set-up international collaboration to create another new project of gene targeting for col4α1 or col4α2, which makes it possible to make mice without type IV collagen.
    We have solved problems of which of the 6 α(IV) chains can make molecules. We raised monoclonal antibodies specific for α(IV) chains. By using these 6 antibodies, we came to the conclusion that there are at least three molecular forms as mentioned above: (α1)2α2, α3α4α5, and (α5)2α6. New biochemical analysis using antibodies that recognize native chains made possible to immunoprecipitate specific molecules when they are digested by pure bacterial collagenase. The products run on SDS-PAGE were analyzed by other monoclonal antibodies that recognize now denatured α-chains by Western blots. This now method concluded that our prediction from immunocytochemistry was indeed true. In other words, there were three (α1)2α2, α3α4α5, and (α5)2α6 molecules. By this method, we analyzed basement membranes from renal glomeruli and bladder and aortic smooth muscle cells.
    Specialized basement membrane from glomeruli was analyzed to contain supramolecular aggregates of (α1)2α2 and α3α4α5 molecules. The function of it is to filter blood to create urine. The similar phenomenon is found in choroid plexus in the brain. We analyzed it and found that it contains also the same compositions as glomerulus.

    researchmap

  • Inhibitory mechanism of vascular endothelial cell proliferation by endostatin

    Grant number:11470274  1999 - 2001

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    NINOMIYA Yoshifumi, YONEZAWA Tomoko, MOMOTA Ryusuke, OOHASHI Toshitaka

      More details

    Grant amount:\13900000 ( Direct expense: \13900000 )

    Under low oxygen condition, expression of collagen XVIII gene by cultured vascular endothelial cells reduced significantly. When it was analyzed by specific monoclonal antibodies, the collagen XVIII protein level was also reduced.
    Endostatin induced regression of chondrosarcoma growth and tumor angiogenesis in vivo. However, endostatin showed no effects on the proliferation and migration of chondrosarcoma cells in vitro. Next, we investigated the interactions between endostatin and endothelial cells in detail. Endostatin inhibited the migration and attachment on collagen I but did not affect proliferation of endothelial cells. Although the migration of endothelial cells was stimulated with angiogenic factors such as basic fibroblast growth factor and vascular endothelial growth factor, endostatin showed similar inhibitory effects on it in the presence or absence of stimulants.
    We came to the conclusion that non-vascular BMs contain predominantly one of the two types; I.e., subepithelial basement membranes contained type XVIII in general, whereas skeletal and cardiac muscles harbored prominently type XV. But basement membranes surrounding smooth muscle cells in vascular tissues contained one or both of them, depending on their locations. Interestingly, continuous or somatic capillaries contained both type XV and type XVIII collagens in their basement membranes; however, fenestrated or specialized capillaries such as glomeruli, liver sinusoids, lung alveoli, and splenic sinusoids expressed only type XVIII chains in their basement membranes, lacking type XV chain. This observation could imply different functions of basement membranes in various tissues and organs that use different mechanisms for the endogenous control of angiogenesis.

    researchmap

  • IV型コラーゲンα5,α6(IV)の組換え体作成により基底膜基質を作る試み

    Grant number:11780623  1999 - 2000

    日本学術振興会  科学研究費助成事業 奨励研究(A)  奨励研究(A)

    百田 龍輔

      More details

    Grant amount:\2100000 ( Direct expense: \2100000 )

    本年度の研究実施計画に基づき研究を行ったところ下記の進展があった。
    CHO細胞がつくるα5(IV)鎖
    11年度の解析からCHO細胞が分泌するIV型コラーゲンはα5(IV)のみであることが明らかになったが、本年度はCHO細胞から分泌されるα5(IV)がどのような形であるかについてさらに検討を行った。
    CHOの培養液を濃縮後、非還元でSDS-PAGEを行ったところモノマーのα5(IV)が検出されるのみで、高分子量の会合体を形成しているものは確認されなかった。さらに未変性のα5(IV)鎖NC1領域を認識する抗体で免疫沈降を行ったがこの抗体で沈殿するα5(IV)は検出されなかった。以上の結果からCHO細胞の発現するα5(IV)はらせん構造をとらないモノマーの形で分泌されていることがわかった。
    α6(IV)鎖cDNA導入を行ったCHO細胞株の解析
    11年度の解析よりα6(IV)鎖全長をコードするcDNA断片を含む発現ベクターを導入したCHO細胞において培養液中と細胞層中でα6(IV)鎖が認められたが、この培養液を濃縮後に未変性のα5(IV)鎖を認識する抗体で免疫沈降を行い、これに対してウェスタンブロットを行ったところα5(IV)鎖とα6(IV)鎖が共存することがわかった。また逆に未変性のα6(IV)NC1領域を認識する抗体を用いて免疫沈降による沈殿物についても同様にα5(IV)鎖とα6(IV)鎖が認められた。
    以上の結果は
    1.α5(IV)鎖だけではらせん会合体を形成しない、
    2.α5(IV)鎖とα6(IV)鎖でらせん会合体を形成する
    ことを反映していることを示唆するものである。
    現在は電子顕微鏡を用いたロータリーシャドウ法で会合体の形態観察を行っている。

    researchmap

  • 新生血管の構築と基底膜コラーゲンの新しい機能

    Grant number:11877121  1999 - 2000

    日本学術振興会  科学研究費助成事業 萌芽的研究  萌芽的研究

    二宮 善文, 米澤 朋子, 百田 龍輔, 大橋 俊孝, 植木 靖好

      More details

    Grant amount:\2000000 ( Direct expense: \2000000 )

    1 血管系特に毛細血管全体におけるXVIIIコラーゲンおよびXVコラーゲン遺伝子の発現
    両コラーゲンは内皮細胞直下基底膜と血管壁に存在する平滑筋細胞周囲の基底膜であることがわかった。毛細血管内皮細胞下基底膜ではXVIII型とXV型を共通に発現するものが認められる.即ち腎や小腸粘膜、皮膚の毛細血管の内皮細胞下基底膜はXVIII型/XV型両者が共存する.しかしこれは組織臓器により異なり、肺胞壁、肝類洞、糸球体基底膜はXVIII型が主であり、他方胎盤、心臓、骨格筋、小腸筋層ではXV型が主であった.しかし、XVIII型/XV型両者の発現のない毛細血管は認められなかった.このように、毛細血管内皮細胞下基底膜のXVIII型とXV型の発現には多様性がある、XVIII型とXV型ともに発現するものが基本だが、類洞や糸球体基底膜のように特殊化した毛細血管ではXVIII型が優位に発現された.
    2 基底膜IV型コラーゲンの新しい機能
    IV型コラーゲンのα鎖は6種類あり、中央にコラーゲン部分、N末(NC2ドメイン)とC末(NC1ドメイン)に非コラーゲンドメインを有する。私達は、これらの6種類のα鎖のNC1ドメインをリコンビナントで作製し、これらを用いて血管内皮細胞の接着性と走化性をみたところ、α2、α3、α6鎖NC1ドメインにそれを抑制する活性が認められた。さらに、血管内皮細胞の接着と走化性はインテグリンαvβ1依存性であることが分かった。鶏胚の系(CAMアッセイ)でbFGFによって誘導された血管新生が、明らかにコントロールとくらべて優位に、リコンビナントα2、α3、α6鎖NC1ドメインによって抑制されるという結果を得た。このことは初めての報告であり、IV型コラーゲンの断片に新しい機能があることが明らかになったという意味で意義のある発見である。

    researchmap

  • Reconstruction of basement membranes aiming at artificial organs

    Grant number:10557113  1998 - 2000

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B).  Grant-in-Aid for Scientific Research (B).

    NINOMIYA Yoshifumi, YONEZAWA Tomoko, MOMOTA Ryusuke, OOHASHI Toshitaka

      More details

    Grant amount:\12900000 ( Direct expense: \12900000 )

    Ultimate goal of the research project is to make artificial organs to insert basement membrane materials between epithelial layers and extracellular matrix. In order to make type IV collagen molecules in vitro, we we did the following investigations :
    * Molecular composition of basement membranes underneath smooth muscle cells varies in different organs, which could be related to specific function of the individual organs.
    * We first isolated and characterized genomic DNA fragments that cover the 5'flanking sequences of COL4A3 and COL4A4 genes, which provided information to delineate the promoter activity for the tissue-specific expression of the six type IV collagen genes.
    * Molecular forms of collagen IV in follicle basement membranes are different in development.
    * We identified interactions between the α2(IV) and ProMMP-9 by immunoprecipitations and blotting.
    * Basement membrane collagen IV molecules diminish in parallel with malignancy and invasiveness when analyzed in tumor progression.
    * Although mRNA expression level of α5 decreased in kidney of a canine Alport case, that of α6 resulted in unchanged. But the protein level of both chains cannot be found any. These results suggest a possibility of a molecular form of α5/α6.
    * Differential expression of collagen IV genes in epithelial basement membranes was analyzed in detail.
    * Here we define a novel function for soluble non-collagenous (NC1) domains of the α2(IV), α3(IV), and α6(IV) chains of human collagen type IV in the regulation of angiogenesis and tumor growth. These NCI domains were shown to regulate endothelial cell adhesion and migration by distinct αv and β1 integrin-dependent mechanisms.
    * In Lmx1b(-/-) mice, expression of both α3 and α4 is strongly diminished in GBM, whereas that of α1, α2 and α5 is unchanged. Moreover, LMX1B binds specifically to a putative enhancer in intron 1 of mouse/human COL4A4 and upregulates reporter constructs containing the enhancer-like sequence.

    researchmap

  • 軟骨細胞分化を誘導する因子のクローニングに関する研究

    Grant number:10877227  1998

    日本学術振興会  科学研究費助成事業 萌芽的研究  萌芽的研究

    二宮 善文, 植木 靖好, 百田 龍輔, 大橋 俊孝

      More details

    Grant amount:\2000000 ( Direct expense: \2000000 )

    本研究は、肢芽発生過程において、軟骨新生を誘導する分化機構を分子の変化として捉えるために、軟骨分化誘導因子をクローニングすることを目的として、現在までに鶏胚の肢芽Stagc20(間葉系細胞)とStage24(軟骨細胞系)からmRNAを抽出し、前者からcDNAライブラリーを作製、後者から引き算し、残りのcDNAをPCRで増幅することにより前者に陽性であって後者に陰性であるクローンを選び出し、これらのクローンについて、そのポリペゾチド構造、遺伝子構造と発現様式を検索し、性質を調べ、軟骨を誘導する機能を有するか否かを調べてきた。取得クローンについて:1)間葉系細胞mRNAと軟骨細胞系mRNAを用いたノザンプロット、2)データーベースによる既報の情報の入手、3)重複久ローン単離により翻訳部位のアミノ酸配列、4)鶏胚全身in situハイブリダイゼーションによる発現パターンの検索、を行ってきた。間葉系細胞と軟骨細胞RNAを抽出し、引算ハイブリダイゼーションPCR法によって、前者に陽性であって後者に陰性であるcDNAクローンを選び出した結果、このなかで興味ある発現パターンを示すいくつかのクローンについて重点的に、ポリペプチド構造と発現部位の同定、他の軟骨特異的遺伝子群との発現時期の比較、等の性質を詳細に検討してきた。得られた複数のクローンが間葉系細胞RNAに強くハイブリダイズし、これらのなかに間葉系細胞を軟骨へと分化を誘導する因子が含まれていることを示唆した。

    researchmap

  • Analysis of specific gene expression of V/XI collagen genes in chondrocyte and non-chondrocyte

    Grant number:09671497  1997 - 1998

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    YOSHIOKA Hidekazu, MATSUO Noritaka, SHIRABE Koumei, MOMOTA Ryusuke, OOHASHI Toshitaka, NINOMIYA Yoshifumi

      More details

    Grant amount:\3200000 ( Direct expense: \3200000 )

    We isolated the 5' flanking region of mouse alphal(XI) collagen gene. As seen in human gene, this promoter has several transcription start sites. Comparison of the sequence of the promoter with consensus regulatory elements showed that there was no TATA or CCAAT box, and that there were several potential Sp1 binding sites.
    Two promoter fragments , 617 bp (short promoter) and 5 kb (long promoter) in size, sharing the same 3 end were subcloned in the upstream of luciferase. And three Sma fragments, 3.5 kb, 3.0 kb and 7.0 kb in the first intron were subeloned in the downstream of the these constructs. We transfected the DNAs into bovine aorta smooth muscle cells ad human chondrosarcoma cells. The luciferase activity of short promoter fragment was higher than that of long promoter in both cells. Three fragments of first intron seemed to depress the activity of promoter.
    To analyze alternative splicing in the acidic region, we made mini genes for the deletion of exon 6A, 6B, 7, 8 and 6A-8, respectively. We transfected these genes in 204 (rhabdomyosarcoma cell), RCS (rat chondrosarcoma) and 293 (kidney cell). In 293 cell, the exon 6A-7-8 is main splice form, and exon 6B is not expressed. In 204 cell, exon 6A-7-8 is the abundant splice form while 7-8 is also present. In the RCS cell, five splice forms are present. In order to analyze the function of the acidic domain in vivo, we have also begun engineering the gene targeting constructs for the deletion of exon 6A and 6B.

    researchmap

  • Functional Analysis of Collagen IV by Gene Targeting

    Grant number:09044308  1997 - 1998

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for international Scientific Research  Grant-in-Aid for international Scientific Research

    NINOMIYA Yoshifumi, MOMOTA Ryusuke, OOHASHI Toshitaka, REINHARD Faessler, CORD Brakebusch, ATTILA Aszodi

      More details

    Grant amount:\5100000 ( Direct expense: \5100000 )

    Col4a6 null mice were created by introducing NeoィイD1RィエD1 gene into exon II of col4a6 gene. We analyzed if the gene is transcribed and translated into α6(IV) polypeptide by in site hybridization, Northern-blot hybridization, and immunohistochemical staining using α chain-specific monoclonal antibodies. The results demonstrated that the α6(IV) polypeptide was not stained at all in any tissues or organs in col4a6 null mice. We also analyzed if the mice have some phenotypes, but we have not detected ant differences between wild type and col4a6 null mice so far.
    We predicted the presence of the three molecular forms of type IV collagen ; [α1(IV)]2α2(IV), α3(IV)α4(IV)α5(IV), and [α5(IV)]2α6α(IV) by use of double staining of the monoclonal antibodies specific for α(IV) chains. The [α1(IV)]2α2(IV) form is present in all basement membranes, whereas the α3(IV)α4(IV)α5(IV) form is localized in glomeruler and alveolar basement membranes and the [α5(IV)]2α6(IV) form is in the Bowman's capsules of the kidney and dermal basement membrane regions. Heterogeneous distribution of the three molecular forms indicated that they may have specific roles in different basement membranes. To describe the biological roles of the molecules, we should think of how the three molecules are incorporated into the supramolecular aggregates of each basement membrane.
    We identified and characterized the breakpoint sequences of the deletion of DNA from a patient of diffuse leiomyomatosis associated with Alport syndrome. The results demonstrated that a deletion eliminates the first coding exon of COL4A5 and the first two of COL4A6. They also showed that the breakpoints share the same sequence, which is in turn closely homologous to the consensuses of topoisomerases I and II. Additional DNA evidence suggested that the male patient is a somatic for the mutation. This study is greatly relevant to the understanding of DL pathogenesis and its etiology.

    researchmap

  • Molecular Mechanism of Esophageal Diffused Leiomyomatosis

    Grant number:09671309  1997 - 1998

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    OOHASHI Toshitaka, UEKI Yasuyoshi, MOMOTA Ryusuke, NINOMIYA Yoshifumi

      More details

    Grant amount:\3000000 ( Direct expense: \3000000 )

    We have investigated the molecular mechanism of diffused leiomyomatosis and the relationship of type IV collagen and smooth muscle cells differentiation and proliferation.
    We identified a DL/AS deletion and first characterization of the break point sequences. The results showed that the breakpoints share the same sequence, which is in turn closly homologous to the consensuses of topoisomerase I and II.The results implicate the genomic rearrangement responsible for the DL/AS phenotype and raise the possibility that there might be a third gene in interveaning sequence III that is involved in the regulation of smooth-muscle-cell proliferation.
    We have generated col4a6 null mice by introducing Neo^r gene into exon2 of col4a6 gene. The mice are fertile and did not show any obvious phenotype. Neither diffused leiomyomatosis in esophagus nor other tumors in smooth muscle organ were found in the mice which lived more than one year. This result exclude the possibility that deletion of col4a6 gene is a cause of DL.
    We are now investigating other possibilities by mating the col4a6 null mice and a transgemic mice which contains the partial genomic fragment delived5 from DL/AS patient. Also exon trapping was performed using a BAC contig which covers huge intron 2 of COL4A6 gene. These study were particularly relevant to the understunding of DL pathogenesis and its etiology.

    researchmap

  • Supramolecular Assembly and Function of New Basement Membrane Collagen Molecules

    Grant number:08457154  1996 - 1997

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    NINOMIYA Yoshifumi, MOMOTA Ryusuke, OOHASHI Toshitaka, YOSHIOKA Hidekatsu

      More details

    Grant amount:\8800000 ( Direct expense: \8800000 )

    To characterize the structure of mouse col4a6 gene, we screened mouse 129 genomic library and isolated a fragment of TSg6, approximately 18.3kb in size. Fine analysis of the genomic fragment and comparison of the structure to that of the cDNA revealed that alternative transcripts that are observed in the human gene do not exist when the mouse is transcribed. In order to create null mouse for col4a6 gene, we manipulated genomic fragment and inserted NcoR gene into the exon 2 of col4a6 gene. The vector was transfected into the R1-Es cells derived from 129v mouse and positive ES cells screened by G418. The positive ES cells were inserted into blastocysts, then the blastocysts were moved uterus of pseudopregnant mice. Chimera mice, heterozygotes and homozygotes are now being checked if any phenotypes are noticed. When homozygote mice were stained by a6 specific monoclonal antibodies, no positive signals was obtained from any tissues, indicating that a6 polypeptide is not expressed in the mice.
    While collagen IV molecules composed of alpha1 and alpha2 chains are broadly distributed, molecules comprising combinations of the other four chains, alpha3-alpha6, are important components of specialized basement membranes. The precise chain composition of triple-helical molecules assembled from the alpha3-alpha6 chains is not entirely clear, but it is hypothesized that alpha3-alpha5 chains and alpha5 and alpha6 chains form heterotrimeric molecules. Several pieces of evidence indicate that alpha3/alpha4/alpha5 molecules and alpha5/alpha6 molecules are components of the basement membrane network.
    We identified an DL/AS deletion and determined the breakpoint wequences of the Japanese case. The results demonstrated that a deletion clininates the first coding exon of COL4A5 and the first two of COL4A6. They also showed that the breakpoints share the same sequence, which is in turn closely homologous to the consensuses of topoisomerases I and II.Additional DNA evidence suggestetd that the male patient is a somatic mosaic for the mutation. Immunohistochemical analysis using alpha chain-specific monoclonal antibodies supportedthis conclusion in that they revealed the absence of the alpha5 (IV) and alpha6 (IV) collagen chains in most but not all of basement membranes of the smooth muscle cell tumor. This study is greatly relevant to the understanding of DL pathogenesis and its etiology.

    researchmap

  • 血液脳関門をマトリックスの分子構築として理解する試み

    Grant number:08877223  1996

    日本学術振興会  科学研究費助成事業 萌芽的研究  萌芽的研究

    二宮 善文, 百田 龍輔, 大橋 俊孝

      More details

    本研究は、選択的透過性を有する脳血液関門がアストロサイトから分泌されるシグナルによって、基底膜を産生する内皮細胞に影響を及ぼし、脳以外の毛細血管周囲基底膜と異なる分子構成による超分子会合体が作り上げられているという考えに基づいて、血液脳関門の本能が細胞外マトリックス分子と細胞間の相互作用で説明することにある。具体的に以下の結果を得た。
    1)脳毛細血管と脳以外の毛細血管(腸間膜)よりRNAを抽出した。これを無細胞系のタンパク翻訳を行うと、多くのポリペプチドが共通のバンドとして認識されるが、一方で、確かにいくつかのペプチドが異なっていることがわかった。これらのバンドが異なる超分子会合体を構成している可能性がある。
    2)ヒト脳毛細血管と脳以外の毛細血管について、α1(IV)-α6(IV)コラーゲン鎖に対する特異的モノクローン抗体を用いて蛍光染色すると、α1(IV)鎖とα2(IV)鎖の存在が明らかになった。脳毛細血管と脳以外の毛細血管について、α(IV)鎖の分布の差は特に認められなかったが、現在さらに詳細の解析を行っている。
    3)脳毛細血管と脳以外の毛細血管より抽出したRNAを鋳型にして、種々のプライマーを用いてDifferential display法を行い、いくつかの異なるバンド認められたので、現在これらをクローニングしている。

    researchmap

  • 肺胞基底膜のIV型コラーゲンの分子種と遺伝子発現

    Grant number:08877098  1996

    日本学術振興会  科学研究費助成事業  萌芽的研究

    百田 龍輔, 吉岡 秀克, 大橋 俊孝

      More details

    Grant amount:\1900000 ( Direct expense: \1900000 )

    今年度、我々の研究は以下のような進展が見られた
    1)IV型コラーゲンα4遺伝子転写産物の発現について
    IV型コラーゲンα4の遺伝子転写産物に関して、我々のこれまでの結果から第1エキソンの異なる2つの転写産物(1と1')が存在することがわかっているが、Northern blot法と定量的RT-PCR法によって、これらの転写産物をそれぞれ独立に検出する系を確立することに成功した。
    これによりIV型コラーゲンα4遺伝子転写産物の発現について、以下のような知見を得た。
    i)2つの転写産物は、基底膜に富む様々な組織において発現している。
    上皮細胞由来の細胞株、基底膜を多く含む組織中で発現が見られた。中でも特に発現が顕著なのは、肺胞の上皮細胞由来の細胞株であった。
    ii)競争的PCR法により転写産物1と1'の転写産物の発現の差について比較を行ったところ、ある種の細胞についてこれらの発現量に100倍もの差が見られた。
    こうした発現の差が、組織特異的プロモーターによるものと考え、更に検討を重ねている。
    2)レポーター遺伝子による、IV型コラーゲンα4遺伝子のプロモーター領域の解析
    α4の転写産物1と1'のそれぞれのプロモーターの活性を見るためのレポーター遺伝子の設計と作製を行った。当初の計画であるルシフェラーゼ遺伝子を用いた測定に先立ち、CAT遺伝子による解析も進行中である。エキソン1'、エキソン1それぞれの上流の2kbをカバーするゲノム断片をCAT遺伝子につないだコンストラクトを作製した。このコンストラクトに基づき、さらに他のコラーゲン遺伝子の例でも見られるようなイントロン1内の転写調節配列の存在の可能性も含めて、数種類のコンストラクトを現在作成中である。

    researchmap

  • V/XI型コラーゲン分子の軟骨・非軟骨細胞における特異的発現機構の解析

    Grant number:08671661  1996

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    吉岡 秀克, 百田 龍輔, 二宮 善文

      More details

    Grant amount:\2100000 ( Direct expense: \2100000 )

    私たちはマウスα1(XI)鎖遺伝子の多様なスプライシングを調べる目的で、エクソン6A、6B、及びC-テロペプチドに対する特異なポリクローナル抗体をウサギに作製した。抗原として合成ペプチドを用いた。得られた抗血清はアフィニテイクロマトグラフィーにて精製し、ELISA法で特異抗体であることを確認した。これらの抗体を用いて、18日目胎児組織における分布を調べた。抗C-テロペプチド抗体による分布は軟骨に強い発現が見られた他に、骨格筋、血管平滑筋、心筋、骨、脳などの非軟骨組織においても分布が見られた。又、抗6B抗体による分布はこの分布と類似していた。しかし、抗6A抗体による分布はこれらとは異なっていた。軟骨では主に肥大軟骨部に見られ、筋組織では動脈平滑筋のみに限局しており、脳においては6Bより強いシグナルが得られた。この分布はα1(V)鎖の分布とも異なっていた。
    又、α1(V)鎖遺伝子の胎児における発現をみる為にRT-PCR法及びNorthern blot法を行った。その結果、肝以外の組織で広く発現が見られた。さらに、18日目胎児マウスの頭蓋骨及び舌組織におけるα1(V)鎖、α2(V)鎖及びα1(XI)鎖遺伝子の発現量をRT-PCR法で調べたところ、組織によりその量比が異なることがわかった。
    このように、V/XI型コラーゲンはα鎖の組み合わせによる分子型の多様性や、スプライシングによるα鎖の機能領域の多様性がみられ、これらは組織或は時期特異的に発現し、機能していることが予想された。

    researchmap

  • Biological function of basement membrane

    Grant number:07044268  1995 - 1996

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for international Scientific Research

    NINOMIYA Yoshifumi, MOMOTA Ryusuke, OOHASHI Toshitaka, APTA Suneel, OLSEN Bjorn, OH Suk Paul, WARMAN Matt

      More details

    Grant amount:\4000000 ( Direct expense: \4000000 )

    1) We have cloned many overlapping cDNAs coding for the mouse alpha6 (IV) collagen chain and determined primary structure of the alpha6 (IV) chain polypeptide.
    2) We raised monoclonal antibodies against amino acid sequences from various parts of the mouse alpha6 (IV) collagen polypeptide chain and used them for the tissue distribution. The results indicated that the tissue distribution for the chain was similar to that of the human alpha6 (IV) collagen chain. Interestingly, the distribution of the paired genes ; col4a1/col4a2, col4a3/col4a4, and col4a5/col4a6 is quite synchronous except for the glomerular basement membranes.
    3) We have some indication that shows alternative splicing in exon 31. Currently, we are characterizing this atypical splicing event in terms of tissue specific expression.
    4) In order to create a knockout mice for col4a6, we made several constructs including the one where the exon 2 was taken out and the foreign gene, Neo^R gene was inserted instead. The construct was transfected into Stem cells by homologous recombination. Currently, we are characterizing knockout mice to detect any phenotypic changes in heterozygotes and homozygotes.

    researchmap

▼display all

 

Class subject in charge

  • Primary Anatomy (2023academic year) special  - その他

  • Practicals: Human Morphology (2023academic year) special  - その他

  • Research Projects: Human Morphology (2023academic year) special  - その他

  • Research Projects and Practicals: Human Morphology I (2023academic year) special  - その他

  • Lecture and Research Projects: Human Morphology I (2023academic year) special  - その他

  • Research Projects and Practicals: Human Morphology II (2023academic year) special  - その他

  • Lecture and Research Projects: Human Morphology II (2023academic year) special  - その他

  • Human Anatomy (2023academic year) special  - その他

  • Human Anatomy (2023academic year) Third semester  - 水4~5

  • Human Anatomy (2023academic year) Third semester  - 水4~5

  • Human Anatomy (2023academic year) Third semester  - 水4~5

  • Human Anatomy (2023academic year) Third semester  - 水4~5

  • Human Gross Anatomy (2023academic year) Concentration  - その他

  • Project-based Learning in Molecular Pathogenesis (2023academic year) special  - その他

  • Human Gross Anatomy (2023academic year) special  - その他

  • Primary Anatomy (2022academic year) special  - その他

  • Research Projects and Practicals: Human Morphology I (2022academic year) special  - その他

  • Lecture and Research Projects: Human Morphology I (2022academic year) special  - その他

  • Research Projects and Practicals: Human Morphology II (2022academic year) special  - その他

  • Lecture and Research Projects: Human Morphology II (2022academic year) special  - その他

  • Human Anatomy (2022academic year) Concentration  - その他

  • Human Anatomy (2022academic year) special  - その他

  • Human Anatomy (2022academic year) Third semester  - 水4~5

  • Human Anatomy (2022academic year) Third semester  - 水4~5

  • Human Anatomy (2022academic year) Third semester  - 水4~5

  • Human Anatomy (2022academic year) Third semester  - 水4~5

  • Human Gross Anatomy (2022academic year) Concentration  - その他

  • Project-based Learning in Molecular Pathogenesis (2022academic year) special  - その他

  • Human Gross Anatomy (2022academic year) special  - その他

  • Primary Anatomy (2021academic year) special  - その他

  • Research Projects and Practicals: Human Morphology I (2021academic year) special  - その他

  • Lecture and Research Projects: Human Morphology I (2021academic year) special  - その他

  • Research Projects and Practicals: Human Morphology II (2021academic year) special  - その他

  • Lecture and Research Projects: Human Morphology II (2021academic year) special  - その他

  • Human Anatomy (2021academic year) Concentration  - その他

  • Human Anatomy (2021academic year) special  - その他

  • Human Anatomy (2021academic year) Third semester  - 水4~5

  • Human Anatomy (2021academic year) Third semester  - 水4~5

  • Human Anatomy (2021academic year) Third semester  - 水4~5

  • Human Anatomy (2021academic year) Third semester  - 水4~5

  • Human Gross Anatomy (2021academic year) Concentration  - その他

  • Project-based Learning in Molecular Pathogenesis (2021academic year) special  - その他

  • Human Gross Anatomy (2021academic year) special  - その他

  • Primary Anatomy (2020academic year) special  - その他

  • Research Projects and Practicals: Human Morphology I (2020academic year) special  - その他

  • Lecture and Research Projects: Human Morphology I (2020academic year) special  - その他

  • Research Projects and Practicals: Human Morphology II (2020academic year) special  - その他

  • Lecture and Research Projects: Human Morphology II (2020academic year) special  - その他

  • Human Anatomy (2020academic year) Concentration  - その他

  • Human Anatomy (2020academic year) special  - その他

  • Human Gross Anatomy (2020academic year) Concentration  - その他

  • Project-based Learning in Molecular Pathogenesis (2020academic year) special  - その他

  • Human Gross Anatomy (2020academic year) special  - その他

▼display all