Updated on 2025/07/16

写真a

 
TSUJI Takehito
 
Organization
Faculty of Environmental, Life, Natural Science and Technology Professor
Position
Professor
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Degree

  • 学術博士（農学） ( 岡山大学 )

Research Interests

  • Animal genetics

  • 動物遺伝学

Research Areas

  • Life Science / Animal life science

  • Life Science / Laboratory animal science

  • Life Science / Animal production science

Education

  • Okayama University    

    - 2001

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  • Okayama University   自然科学研究科  

    - 2001

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    Country： Japan

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  • Okayama University    

    - 1994

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  • Okayama University   農学部   総合農業科

    - 1994

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    Country： Japan

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Research History

  • - 岡山大学 環境生命科学研究科 准教授

    2012

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  • - Associate Professor

    2012

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  • 岡山大学 自然科学研究科 准教授

    2010 - 2012

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  • Associate Professor

    2010 - 2012

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  • - Associate Professor,Graduate School of Environmental and life Science,Okayama University

    2010

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  • - 岡山大学環境生命科学研究科 准教授

    2010

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  • Research Associate

    2002 - 2010

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  • Okayama University   Faculty of Agriculture

    2002 - 2010

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  • Research Associate

    1999 - 2002

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  • Okayama University   Dental School

    1999 - 2002

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Professional Memberships

 

Papers

  • White coat color in Vietnamese native buffalo is attributed to the LINE1 insertion in ASIP

    Thuy Thanh Nguyen, Quan Viet Le, Van Huu Nguyen, Hai Thanh Duong, Takehito Tsuji

    Tropical Animal Health and Production   57 ( 2 )   2025.2

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    Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

    Abstract

    The coat color of the swamp buffalo is commonly dark gray, while the white coat color variant, which may have potential heat stress advantages, is also present in some Asian countries, including Vietnam. This study analyzed the most likely candidate genes, ASIP and TYR, responsible for the white coat color of Vietnamese native buffaloes. We found that LINE1 insertion in ASIP, a mutation previously reported in white swamp buffalo from other Asian countries, was exclusively found in white Vietnamese buffalo. Moreover, significantly higher expression of ASIP was confirmed in the hair follicles of white buffalo. On the other hand, no variants associated with the white phenotype were found in TYR. These findings indicate that the LINE1 insertion in ASIP is responsible for the white coat color in Vietnamese native buffalo, and that provides a crucial step towards their utilization and improved productivity in Vietnam.

    DOI： 10.1007/s11250-025-04309-7

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    Other Link： https://link.springer.com/article/10.1007/s11250-025-04309-7/fulltext.html

  • Investigation of SNPs associated with reproductive and body growth traits in Vietnamese and Nepalese native buffaloes

    Nguyen Thuy Thanh, Tetsuo Kunieda, Shah Manoj Kumar, Le Thu Nu Anh, Nguyen Van Huu, Koichiro Ushijima, Mayuko Nagae, Takehito Tsuji

    The Journal of Animal Genetics   53 ( 1 )   3 - 3   2025.1

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    Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Animal Breeding and Genetics  

    DOI： 10.5924/abgri.53.3

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  • Association of a non-synonymous variant of MLH3 gene involved in meiotic recombination with conception rate of Japanese Black cattle

    Thu Nu Anh Le, Trung Ba Nguyen, Takehito Tsuji, Takayuki Ibi, Shinji Sasaki, Tetsuo Kunieda

    Animal Production Science   64 ( 17 )   2024.11

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    Publishing type：Research paper (scientific journal)   Publisher：CSIRO Publishing  

    Context Conception rate, which is an important parameter to evaluate female fertility, has been gradually decreasing in Japanese Black cattle during the past decades. Meiosis is an essential biological process in gamete formation and meiotic failure could be a cause of infertility or reduced fertility in mammals. Of note, single nucleotide polymorphisms (SNPs) in the MLH3 gene involved in meiotic recombination were reported to affect the genome-wide recombination rate of meiosis in cattle. Aims The present study was conducted to investigate the association of SNPs in the MLH3 gene with conception rate in Japanese Black cattle. Methods On the basis of the reproductive data of 2045 Japanese Black cattle born from 1990 to 2009, we selected two groups of the reproductive females with high conception rate (n = 103) and low conception rate (n = 109). Then, we genotyped the SNPs in MLH3 gene in both reproductive groups by sequencing and polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). The significant association of SNPs with conception rate in Japanese Black cattle was estimated by Fisher’s exact test, by using R commander. Key results We found the presence of five SNPs of MLH3 gene in Japanese Black cattle, including non-synonymous variants of MLH3 N408S that has been reported to be involved in meiotic recombination rate and MLH3 S591G newly identified in the present study. Comparison of genotype distributions and allele frequencies of the MLH3 N408S between high and low conception-rate groups indicated a significant difference between these groups, suggesting that this SNP is associated with conception rate. However, there is no significant difference between MLH3 S591G with conception rate in Japanese Black cattle. Conclusions We found a significant association between a non-synonymous variant of MLH3 N408S and conception rate, suggesting that this SNP can be a potential marker for selecting the Japanese Black cattle for improving fertility. We also identified a novel non-synonymous variant of MLH3 S591G in Japanese Black cattle. Implications These findings will be informative for future marker-assisted selection to improve the fertility of Japanese Black cattle. This is the first report of a genetic variant implicated in meiotic recombination being linked to female fertility in cattle.

    DOI： 10.1071/an23418

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  • A nonsense mutation in mouse Adamtsl2 causes uterine hypoplasia and an irregular estrous cycle. International journal

    Yuka Iwanaga, Kaori Tsuji, Ayaka Nishimura, Kouji Tateishi, Misa Kakiuchi, Takehito Tsuji

    Mammalian genome : official journal of the International Mammalian Genome Society   34 ( 4 )   559 - 571   2023.12

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

    The spontaneous mutation stubby (stb) in mice causes chondrodysplasia and male infertility due to impotence through autosomal recessive inheritance. In this study, we conducted linkage analysis to localize the stb locus within a 1.6 Mb region on mouse chromosome 2 and identified a nonsense mutation in Adamtsl2 of stb/stb mice. Histological analysis revealed disturbed endochondral ossification with a reduced hypertrophic chondrocyte layer and stiff skin with a thickened dermal layer. These phenotypes are similar to those observed in humans and mice with ADAMTSL2/Adamtsl2 mutations. Moreover, stb/stb female mice exhibited severe uterine hypoplasia at 5 weeks of age and irregular estrous cycles at 10 weeks of age. In normal mice, Adamtsl2 was more highly expressed in the ovary and pituitary gland than in the uterus, and this expression was decreased in stb/stb mice. These findings suggest that Adamtsl2 may function in these organs rather than in the uterus. Thus, we analyzed Gh expression in the pituitary gland and plasma estradiol and IGF1 levels, which are required for the development of the female reproductive tract. There was no significant difference in Gh expression and estradiol levels, whereas IGF1 levels in stb/stb mice were significantly reduced to 54-59% of those in +/+ mice. We conclude that Adamtsl2 is required for the development of the uterus and regulation of the estrous cycle in female mice, and decreased IGF1 may be related to these abnormalities.

    DOI： 10.1007/s00335-023-10016-1

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  • Genetic variants of RNF212 involved in meiotic recombination rate and its relation with conception rate in Japanese Black cattle

    Thu Nu Anh LE, Trung Ba NGUYEN, Ripon Chandra PAUL, Yu OKUDA, Takehito TSUJI, Takayuki IBI, Shinji SASAKI, Tetsuo KUNIEDA

    The Journal of Animal Genetics   49 ( 1 )   13 - 17   2021

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    Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Animal Breeding and Genetics  

    DOI： 10.5924/abgri.49.13

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  • Genetic characterization of Kushum horses in Kazakhstan based on haplotypes of mtDNA and Y chromosome, and genes associated with important traits of the horses

    Trung B. NGUYEN, Ripon C. PAUL, Yu OKUDA, Thu N. A. LE, Phuong T. K. PHAM, Kushaliye J. KAISSAR, Akhmedenov KAZHMURAT, Sarsenova BIBIGUL, Meirat BAKHTIN, Polat KAZYMBET, Suleimenov Zh MARATBEK, Alikhan MELDEBEKOV, Masahide NISHIBORI, Takayuki IBI, Takehito TSUJI, Tetsuo KUNIEDA

    Journal of Equine Science   31 ( 3 )   35 - 43   2020

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japan Society of Equine Science  

    The Kushum is a relatively new breed of horses in Kazakhstan that was established in the middle of the 20th century through a cross between mares of Kazakhstan local horses and stallions of Thoroughbred, Trotter, and Russian Don breeds to supply military horses. To reveal the genetic characteristics of this breed, we investigated haplotypes of mitochondrial DNA (mtDNA) and single-nucleotide polymorphisms of the Y chromosome, as well as genotypes of five functional genes associated with coat color, body composition, and locomotion traits. We detected 10 mtDNA haplotypes that fell into 8 of the 17 major haplogroups of horse mtDNA, indicating a unique haplotype composition with high genetic diversity. We also found two Y-chromosomal haplotypes in Kushum horses, which likely originated from Trotter and/or Don breeds. The findings regarding the mtDNA and Y-chromosomal haplotypes are concordant with the documented maternal and paternal origins of the Kushum horses. The allele frequencies of ASIP, MC1R, and MATP associated with coat color were consistent with the coat color variations of Kushum horses. The allele frequencies of MSTN associated with endurance performance and those of DMRT3 associated with gait suggested that the observed allele frequencies of these genes were the result of selective breeding for these traits. As a result of this study, we were able to obtain useful information for a better understanding of the origin and breeding history of the Kushum horse breed using molecular markers.

    DOI： 10.1294/jes.31.35

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  • A mutation in the nuclear pore complex gene Tmem48 causes gametogenesis defects in skeletal fusions with sterility (sks) mice. Reviewed

    Akiyama K, Noguchi J, Hirose M, Kajita S, Katayama K, Khalaj M, Tsuji T, Fairfield H, Byers C, Reinholdt L, Ogura A, Kunieda T

    The Journal of biological chemistry   288 ( 44 )   31830 - 31841   2013.11

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  • Screening of TMEM48 binding proteins in testis by yeast two-hybrid method

    KAJITA Shimpei, AKIYAMA Kouyou, TSUJI Takehito, KUNIEDA Tetsuo

    The Journal of animal genetics   41 ( 2 )   77 - 86   2013

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    Language：Japanese   Publisher：Japanese Society of Animal Breeding and Genetics  

    Tmem48 encodes a nuclear membrane protein comprising the nuclear pore complex and a mutation in this gene is responsible for the gametogenesis defects and skeletal malformations in the sks mutant mice. Since a nucleotide substitution in the Tmem48 gene results in arrest of meiosis that causes defective spermatogenesis in the sks mice, the function of nuclear pore complex thorough TMEM48 is suggested to be essential for mammalian gametogenesis. In the present study, therefore, we attempted to identify proteins that have potentially important functions in mammalian gametogenesis by screening of the testis expressing proteins that bind to TMEM48 using yeast two-hybrid method. As a result of the screening, we identified total of 31 proteins which are suggested to interact with TMEM48. The expression analysis of these genes in various mouse tissues indicated specific expressions of Dynll2, Pabpc2, Spink2, Txnl4b genes in the testis. Then, we examined expressions of these genes during the first wave of spermatogenesis at new born stages, in which synchronized spermatogenesis is observed in all seminiferous tubules, and found that these genes express at specific stages of meiosis during spermatogenesis. These findings suggested that the proteins encoded by these genes play essential roles in mammalian gametogenesis.

    DOI： 10.5924/abgri.41.77

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  • 卵母細胞の減数分裂制御機構におけるC型ナトリウム利尿ペプチドの役割

    辻 岳人, 清須 千代, 秋山 耕陽, 国枝 哲夫

    日本繁殖生物学会 講演要旨集   106   P - 75-P-75   2013

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    Language：Japanese   Publisher：日本繁殖生物学会  

    【目的】卵巣内の卵母細胞は減数分裂第一分裂前期の状態を維持し，LHサージにより再開する。C型ナトリウム利尿ペプチド(CNP)は，受容体(NPR2)により合成されたcGMPを介して減数分裂の停止維持に関わることが後期胞状卵胞でのみ報告されている。本研究では，CNPシグナルによる減数分裂抑制効果がより早期卵胞で機能しているか調べ，さらに，LHによる減数分裂再開への関与についても解析をおこなった。 【方法】 1．Npr2遺伝子に機能欠失型の突然変異をもつマウスについて，25日齢の卵巣切片を作成し，卵母細胞における核の状態を観察した。2．25日齢の正常マウスとPMSG，hCGを投与した正常マウス卵巣におけるCNP (Nppc) 遺伝子および受容体(Npr2)遺伝子の発現パターンを解析した。3．正常マウス卵巣より単離した顆粒膜細胞を培養し，アンフィレギュリンおよびEGFレプター阻害剤によるNppc遺伝子の発現量への影響を解析した。【結果】 Npr2遺伝子に変異をもつマウスの卵巣では，前胞状卵胞までの卵母細胞は正常マウスと同様にGV期であったが，初期胞状卵胞以降の半数以上の卵母細胞では染色体が観察され，減数分裂が異常に再開していた。初期胞状卵胞では卵胞外側および卵母細胞周囲の顆粒膜細胞にNppcおよびNpr2遺伝子はそれぞれ発現し，PMSG投与48時間後の胞状卵胞では，顆粒膜細胞と卵丘細胞において発現していた。また，Nppc遺伝子の発現はhCG投与後に顕著に減少した。培養顆粒膜細胞では，Nppc遺伝子の発現量はアンフィレギュリン添加により減少し，EGFレセプター阻害剤の前処理により発現量の低下は認められなかった。以上の結果より，CNP/NPR2シグナルは，胞状卵胞期以降の卵母細胞の減数分裂の停止維持に関わる体細胞由来の必須因子であることが示された。また，LHサージによる減数分裂再開には，CNPによる減数分裂停止維持効果の低下が関与している可能性が示された。

    DOI： 10.14882/jrds.106.0.P-75.0

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  • Exclusion of NEU1 and PPGB from candidate genes for a lysosomal storage disease in Japanese Black cattle. Reviewed

    Masoudi AA, Yamato O, Yoneda K, Tsuji T, Mikami O, Kunieda T

    Animal science journal = Nihon chikusan Gakkaiho   80 ( 5 )   611 - 615   2009.10

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  • Characterization and linkage mapping of an ENU-induced mutant mouse with defective spermatogenesis. Reviewed

    Asano Y, Akiyama K, Tsuji T, Takahashi S, Noguchi J, Kunieda T

    Experimental animals   58 ( 5 )   525 - 532   2009.10

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    Language：English   Publisher：Japanese Association for Laboratory Animal Science  

    repro23 is an autosomal recessive mutation of the mouse generated by the N-ethyl-N-nitrosourea (ENU)-induced mutagenesis program at The Jackson Laboratory. The repro23/repro23 homozygous mouse shows male-specific infertility caused by defective spermatogenesis. In the present study, we investigated the testicular pathology of the affected mouse and performed linkage analysis to determine the chromosomal localization of the repro23 locus. Histological examination of the affected testis showed that the seminiferous epithelium of the repro23/repro23 mice contained spermatogonia and early stage spermatocytes, but no spermatids or spermatozoa. Immunohistochemical staining for Hsc70t, a spermatid specific protein, confirmed the absence of elongating spermatids. These findings indicated interruption of the spermatogenesis during meiosis in the repro23/repro23 mouse. By linkage analysis using 137 affected mice of F2 progeny obtained from crosses between repro23/repro23 female and JF1/Ms (+/+) male mice, the repro23 locus was mapped to 2.2-Mb region of mouse chromosome 7. Although this region contains several potential candidate genes for the repro23 mutation, no gene already identified as a cause of defective speramatogenesis was in this region. Therefore, the gene responsible for the repro23 mutation is suggested to be a novel gene which plays an essential role in mammalian spermatogenesis.<br>

    DOI： 10.1538/expanim.58.525

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    Other Link： https://jlc.jst.go.jp/DN/JALC/00340059676?from=CiNii

  • Characterization of the chromosomal inversion associated with the Koa mutation in the mouse revealed the cause of skeletal abnormalities. Reviewed

    Katayama K, Miyamoto S, Furuno A, Akiyama K, Takahashi S, Suzuki H, Tsuji T, Kunieda T

    BMC genetics   10   60   2009.9

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  • Polymerase chain reaction-restriction fragment length polymorphism method for identifying carriers of hemophilia A in Japanese brown cattle Reviewed

    Maryam Khalaj, Abdol Rahim Abbasi, Takehito Tsuji, Yasuo Moritomo, Kenichi Shimojo, Tetsuo Kunieda

    Animal Science Journal   77 ( 1 )   122 - 125   2006.2

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

    DOI： 10.1111/j.1740-0929.2006.00329.x

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  • FKBP6 deficiency in rats disorders homologous chromosome pairing, but not sex chromosome inactivation.

    J Noguchi, T Tsuji, NWK Karja, M Fahrudin, M Ozawa, K Ohnuma, K Kikuchi, H Kaneko, T Kunieda

    BIOLOGY OF REPRODUCTION   162 - 162   2005

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  • Loss of Function of Evc2 in Dental Mesenchyme Leads to Hypomorphic Enamel

    H. Zhang, H. Takeda, T. Tsuji, N. Kamiya, T. Kunieda, Y. Mochida, Y. Mishina

    JOURNAL OF DENTAL RESEARCH   96 ( 4 )   414 - 422   2017.4

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  • Elevated Fibroblast Growth Factor Signaling Is Critical for the Pathogenesis of the Dwarfism in Evc2/Limbin Mutant Mice

    Honghao Zhang, Nobuhiro Kamiya, Takehito Tsuji, Haruko Takeda, Greg Scott, Sudha Rajderkar, Manas K. Ray, Yoshiyuki Mochida, Benjamin Allen, Veronique Lefebvre, Irene H. Hung, David M. Ornitz, Tetsuo Kunieda, Yuji Mishina

    PLOS GENETICS   12 ( 12 )   2016.12

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  • Ellis Van Creveld2 is Required for Postnatal Craniofacial Bone Development

    Mohammed K. Badri, Honghao Zhang, Yoshio Ohyama, Sundharamani Venkitapathi, Nobuhiro Kamiya, Haruko Takeda, Manas Ray, Greg Scott, Takehito Tsuji, Tetsuo Kunieda, Yuji Mishina, Yoshiyuki Mochida

    ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY   299 ( 8 )   1110 - 1120   2016.8

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  • Expression of Evc2 in craniofacial tissues and craniofacial bone defects in Evc2 knockout mouse

    Mohammed K. Badri, Honghao Zhang, Yoshio Ohyama, Sundharamani Venkitapathi, Ahmed Alamoudi, Nobuhiro Kamiya, Haruko Takeda, Manas Ray, Greg Scott, Takehito Tsuji, Tetsuo Kunieda, Yuji Mishina, Yoshiyuki Mochida

    ARCHIVES OF ORAL BIOLOGY   68   142 - 152   2016.8

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  • Homeobox family Hoxc localization during murine palate formation

    Azumi Hirata, Kentaro Katayama, Takehito Tsuji, Hideto Imura, Nagato Natsume, Toshio Sugahara, Tetsuo Kunieda, Hiroaki Nakamura, Yoshinori Otsuki

    CONGENITAL ANOMALIES   56 ( 4 )   172 - 179   2016.7

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  • Genetic characterization of a rare line of Japanese Black cattle in Okayama prefecture

    YONEDA Kazuhiro, OKUDA Yu, Siqintuya, NISHIMAKI Takahiro, MATSUMOTO Hirokazu, MIYAZAKI Yoshiyuki, IBI Takayuki, TSUJI Takehito, KUNIEDA Tetsuo

    Nihon Chikusan Gakkaiho   87 ( 1 )   1 - 10   2016

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    Language：Japanese   Publisher：Japanese Society of Animal Science  

    The effective population size of Japanese Black cattle has been significantly reduced and maintaining the genetic diversity of the population is important for breeding of Japanese Black cattle. For the conservation of genetic diversity, use of different lines of the breed with unique genetic characteristics for breeding, instead of intensive use of sires of few particular lines as common in current breeding of Japanese Black cattle, will be effective to prevent the genetic homogenization of the population. In the present study, we performed genetic characterization a population of a rare line of Japanese Black cattle, which has been originated from ancestral &quot;Tsuru-ushi&quot; in Okayama prefecture. By using microsatellite markers, allelic richness, and average observed and expected heterozygosity are 3.48, 0.514, and 0.511, respectively, and these values were lower than those of most of the Japanese Black cattle local subpopulations compared. The result of the clustering analyses indicated that the animals of the rare line formed a single group with a cluster that was clearly distinguished from the other populations. Sequence analysis of mitochondrial D-loop region revealed that only two haplotypes were observed in the population and 80% of the animals in the population possess a single haplotype. However, the other haplotype was a novel unique haplotype that has not been reported in cattle. Genotyping of six genes associated with important traits revealed that &lt;i&gt;SREBP1&lt;/i&gt; and &lt;i&gt;NCAPG&lt;/i&gt; loci were fixed for a single allele in the population and more than 90% of animals possess an allele of &lt;i&gt;MC1R&lt;/i&gt; locus. These findings indicated that while genetic diversity of the population of the rare line is lower than those of the Japanese Black cattle local subpopulations, this population has unique genetic characteristics that were distinguished from the other populations and, therefore, the rare line is important for maintaining genetic diversity of Japanese Black cattle.

    DOI： 10.2508/chikusan.87.1

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  • Generation of Evc2/Limbin Global and Conditional KO Mice and Its Roles during Mineralized Tissue Formation

    Honghao Zhang, Haruko Takeda, Takehito Tsuji, Nobuhiro Kamiya, Sudha Rajderkar, Ke'Ale Louie, Crystal Collier, Greg Scott, Manas Ray, Yoshiyuki Mochida, Vesa Kaartinen, Tetsuo Kunieda, Yuji Mishina

    GENESIS   53 ( 9 )   612 - 626   2015.9

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  • Non-synonymous FGD3 Variant as Positional Candidate for Disproportional Tall Stature Accounting for a Carcass Weight QTL (CW-3) and Skeletal Dysplasia in Japanese Black Cattle

    Akiko Takasuga, Kunio Sato, Ryouichi Nakamura, Yosuke Saito, Shinji Sasaki, Takehito Tsuji, Akio Suzuki, Hiroshi Kobayashi, Tamako Matsuhashi, Koji Setoguchi, Hiroshi Okabe, Toshitake Ootsubo, Ichiro Tabuchi, Tatsuo Fujita, Naoto Watanabe, Takashi Hirano, Shota Nishimura, Toshio Watanabe, Makio Hayakawa, Yoshikazu Sugimoto, Takatoshi Kojima

    PLOS GENETICS   11 ( 8 )   e1005433   2015.8

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  • 岡山県のジャージー集団におけるBCO2遺伝子の新たな変異と脂肪淡黄色との関連性

    清水佑起, 石倉健一, 揖斐隆之, 国枝哲夫, 辻岳人

    日本畜産学会大会講演要旨   119th   115   2015.3

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    Language：Japanese  

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  • 骨格異常と枝肉重量QTL(CW‐3)は体高の増加と胸幅の減少によって説明される

    高須賀晶子, 齊藤陽介, 佐藤邦雄, 中村亮一, 小林宙, 松橋珠子, 鈴木晶夫, 瀬戸口浩二, 佐々木慎二, 早川磨紀男, 辻岳人, 小島孝敏

    日本畜産学会大会講演要旨   119th   171 - 171   2015.3

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    Language：Japanese   Publisher：(公社)日本畜産学会  

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  • 精子頭部及び尾部の形成異常を呈するENU誘発ミュータントマウス(repro20,21)の原因遺伝子の同定

    松本大和, 奥田ゆう, 藤原靖浩, 秋山耕陽, 辻岳人, 国枝哲夫

    J Reprod Dev   60 ( Suppl Japanese Issue )   J63 - j63   2014.8

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    Language：Japanese   Publisher：(一社)日本繁殖生物学会  

    DOI： 10.14882/jrds.107.0_OR1-2

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  • Role of Ellis-van Creveld syndrome2 (Evc2) in craniofacial development

    Mohammed K. Badri, Honghao Zhang, Nobuhiro Kamiya, Takehito Tsuji, Tetsuo Kunieda, Yuji Mishina, Yoshiyuki Mochida

    AMERICAN JOURNAL OF MEDICAL GENETICS PART A   164 ( 8 )   1876 - 1877   2014.8

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    Language：English   Publishing type：Research paper, summary (international conference)  

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  • 不妊を呈するENU誘発ミュータントマウス(repro57)雌の表現型解析

    筑紫未津穂, 藤原靖浩, 松本大和, 秋山耕陽, 辻岳人, 国枝哲夫

    日本実験動物学会総会講演要旨集   61st   261   2014.5

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  • Characterization of the Skeletal Fusion with Sterility (sks) Mouse Showing Axial Skeleton Abnormalities Caused by Defects of Embryonic Skeletal Development

    Kouyou Akiyama, Kentaro Katayama, Takehito Tsuji, Tetsuo Kunieda

    EXPERIMENTAL ANIMALS   63 ( 1 )   11 - 19   2014.1

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  • 口之島牛集団における経済形質、遺伝性疾患および毛色に関連する遺伝子の対立遺伝子頻度とその分布

    動物遺伝育種研究   42 ( 1 )   11 - 19   2014

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  • Phenotypic Characterization of Ggt1dwg/dwg Mice, a Mouse Model for Hereditary gamma-Glutamyl Transferase Deficiency

    Kaoru Yamada, Takehito Tsuji, Tetsuo Kunieda

    EXPERIMENTAL ANIMALS   62 ( 2 )   151 - 157   2013.4

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  • Heparanase Localization during Palatogenesis in Mice

    Azumi Hirata, Kentaro Katayama, Takehito Tsuji, Nagato Natsume, Toshio Sugahara, Yuichi Koga, Kazufumi Takano, Yoshinori Otsuki, Hiroaki Nakamura

    BIOMED RESEARCH INTERNATIONAL   2013

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  • CNP/NPR2 signaling maintains oocyte meiotic arrest in early antral follicles and is suppressed by EGFR-mediated signaling in preovulatory follicles

    Takehito Tsuji, Chiyo Kiyosu, Kouyou Akiyama, Tetsuo Kunieda

    MOLECULAR REPRODUCTION AND DEVELOPMENT   79 ( 11 )   795 - 802   2012.11

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  • NPPC/NPR2 signaling is essential for oocyte meiotic arrest and cumulus oophorus formation during follicular development in the mouse ovary

    Chiyo Kiyosu, Takehito Tsuji, Kaoru Yamada, Shimpei Kajita, Tetsuo Kunieda

    REPRODUCTION   144 ( 2 )   187 - 193   2012.8

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  • Skeletal Analysis of the Long Bone Abnormality (lbab/lbab) Mouse, A Novel Chondrodysplastic C-Type Natriuretic Peptide Mutant

    Eri Kondo, Akihiro Yasoda, Takehito Tsuji, Toshihito Fujii, Masako Miura, Naotestu Kanamoto, Naohisa Tamura, Hiroshi Arai, Tetsuo Kunieda, Kazuwa Nakao

    CALCIFIED TISSUE INTERNATIONAL   90 ( 4 )   307 - 318   2012.4

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  • A missense mutation of the Dhh gene is associated with male pseudohermaphroditic rats showing impaired Leydig cell development

    Yasuhiro Kawai, Junko Noguchi, Kouyou Akiyama, Yuriko Takeno, Yasuhiro Fujiwara, Shimpei Kajita, Takehito Tsuji, Kazuhiro Kikuchi, Hiroyuki Kaneko, Tetsuo Kunieda

    REPRODUCTION   141 ( 2 )   217 - 225   2011.2

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  • Gastrointestinal Tract Disorder in Natriuretic Peptide Receptor B Gene Mutant Mice

    Chizuru Sogawa, Asaki Abe, Takehito Tsuji, Mitsuru Koizumi, Tsuneo Saga, Tetsuo Kunieda

    AMERICAN JOURNAL OF PATHOLOGY   177 ( 2 )   822 - 828   2010.8

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  • The Roles of Syntaxin2/Epimorphin (Stx2/Epim) in Progression of Meiosis During Spermatogenesis

    Yasuhiro Fujiwara, Kouyou Akiyama, Yuka Asano, Takehito Tsuji, Junko Noguchi, Tetsuo Kunieda

    BIOLOGY OF REPRODUCTION   136 - 136   2010

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  • Cloning of Candidate Genes for TMEM48 Binding Proteins, Which Could Be Involved in Gametogenesis

    Shimpei Kajita, Kouyou Akiyama, Michiko Hirose, Narumi Ogonuki, Atsuo Ogura, Takehito Tsuji, Tetsuo Kunieda

    BIOLOGY OF REPRODUCTION   136 - 136   2010

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  • Characterization of the dwg mutations: dwg and dwg (Bayer) are new mutant alleles of the Ggt1 gene

    Takehito Tsuji, Kaoru Yamada, Tetsuo Kunieda

    MAMMALIAN GENOME   20 ( 11-12 )   711 - 719   2009.12

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  • A mutation of the WFDC1 gene is responsible for multiple ocular defects in cattle

    Abdol Rahim Abbasi, Maryam Khalaj, Takehito Tsuji, Muki Tanahara, Kazuyuki Uchida, Yoshikazu Sugimoto, Tetsuo Kunieda

    GENOMICS   94 ( 1 )   55 - 62   2009.7

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  • Hypomorphic mutation in mouse Nppc gene causes retarded bone growth due to impaired endochondral ossification

    Takehito Tsuji, Eri Kondo, Akihiro Yasoda, Masataka Inamoto, Chiyo Kiyosu, Kazuwa Nakao, Tetsuo Kunieda

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   376 ( 1 )   186 - 190   2008.11

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  • Leydig cell hyperplasia in an ENU-induced mutant mouse with germ cell depletion

    Maryarn Khalaj, Abdol Rahim Abbasi, Ryo Nishimura, Kouyou Akiyama, Takehito Tsuji, Junko Noguchi, Kiyoshi Okuda, Tetsuo Kunieda

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   54 ( 3 )   225 - 228   2008.6

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  • A new ENU-induced mutant mouse with defective spermatogenesis caused by a nonsense mutation of the syntaxin 2/Epimorphin (Stx2/Epim) gene

    Kouyou Akiyama, Shiho Akimaru, Yuka Asano, Maryam Khalaj, Chiyo Kiyosu, Ali Akbar Masoudi, Sakino Takahashi, Kentaro Katayama, Takehito Tsuji, Junko Noguchi, Tetsuo Kunieda

    JOURNAL OF REPRODUCTION AND DEVELOPMENT   54 ( 2 )   122 - 128   2008.4

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  • Linkage mapping of the locus responsible for forelimb-girdle muscular anomaly of Japanese black cattle on bovine chromosome 26

    A. A. Masoudi, K. Uchida, K. Yokouchi, K. Ohwada, A. R. Abbasi, T. Tsuji, T. Watanabe, T. Hirano, Y. Sugimoto, T. Kunieda

    ANIMAL GENETICS   39 ( 1 )   46 - 50   2008.2

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  • Suppressed recombination on mouse chromosome 15 defined regions of chromosomal inversions associated with koala (Koa) and hairy ears (Eh) mutations

    Kentaro Katayama, Aki Furuno, Sayaka Miyamoto, Miyuki Nakamura, Izurni Ojika, Yusuke Shinkai, Kouyou Akiyama, Takehito Tsuji, Tetsuo Kunieda

    EXPERIMENTAL ANIMALS   57 ( 1 )   73 - 77   2008.1

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  • Short-limbed dwarfism: slw is a new allele of Npr2 causing chondrodysplasia

    Chizuru Sogawa, Takehito Tsuji, Yusuke Shinkai, Kentaro Katayama, Tetsuo Kunieda

    JOURNAL OF HEREDITY   98 ( 6 )   575 - 580   2007.9

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  • Limbinノックアウトマウスは軟骨内骨化の異常による骨成長遅延を呈する

    稲本政隆, 辻岳人, 神谷宣広, 三品裕司, 国枝哲夫

    日本骨代謝学会学術集会プログラム抄録集   25th   245 - 245   2007.6

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  • Characterization of chromosomal inversion of the mouse hairy ears (Eh) mutation associated with cleft palate

    Kentaro Katayama, Aki Furuno, Kouyou Akiyama, Takehito Tsuji, Tetsuo Kunieda

    MAMMALIAN GENOME   18 ( 4 )   246 - 254   2007.4

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  • A mutation causing abnormal splicing of Tmem48/NdC1 gene is responsible for impaired gametogenesis in SKS mutant mouse.

    Kouyou Akiyama, Junko Noguchi, Mai Kanaeda, Takehito Tsuji, Tetsuo Kunieda

    BIOLOGY OF REPRODUCTION   90 - 90   2007

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  • CNP/NPRB plays an important role in follicular, development and oocyte maturation.

    Chiyo Iyosu, Takehito Tsuji, Tetsuo Kunieda

    BIOLOGY OF REPRODUCTION   115 - 116   2007

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  • Limbinのノックアウトマウスは軟骨低形成,短肢症を呈する~Ellis‐van Creveld syndromeの疾患モデルマウスの作成~

    神谷宣広, 竹田晴子, 辻岳人, 国枝哲夫, 三品裕司

    日本軟骨代謝学会プログラム・抄録集   20th   80   2007

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  • A deletion in the endothelin-B receptor gene is responsible for the Waardenburg syndrome-like phenotypes of WS4 mice

    Shin Ohtani, Yusuke Shinkai, Akio Horibe, Kentaro Katayama, Takehito Tsuji, Yoshibumi Matsushima, Masayoshi Tachibana, Tetsuo Kunieda

    EXPERIMENTAL ANIMALS   55 ( 5 )   491 - 495   2006.10

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  • Reduced expression of the endothelin receptor type B gene in piebald mice caused by insertion of a retroposon-like element in intron 1

    T Yamada, S Ohtani, T Sakurai, T Tsuji, T Kunieda, M Yanagisawa

    JOURNAL OF BIOLOGICAL CHEMISTRY   281 ( 16 )   10799 - 10807   2006.4

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  • A PCR-RFLP method for identifying carriers of Hemophilia A in Japanese brown cattle

    Maryam Khalaj, Abdol Rahim Abbasi, Takehito Tsuji, Yasuo Moritomo, Kenichi Shimojo, Tetsuo Kunieda

    Anim. Sci. J.   2006

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  • Linkage mapping of the locus responsible for congenital multiple ocular defects in cattle on bovine Chromosome 18

    AR Abbasi, N Ihara, T Watanabe, M Khalaj, T Tsuji, Y Sugimoto, T Kunieda

    MAMMALIAN GENOME   16 ( 9 )   731 - 737   2005.9

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  • An insertion mutation of the bovine F11 gene is responsible for factor XI deficiency in Japanese black cattle

    M Kunieda, T Tsuji, AR Abbasi, M Khalaj, M Ikeda, K Miyadera, H Ogawa, T Kunieda

    MAMMALIAN GENOME   16 ( 5 )   383 - 389   2005.5

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  • A loss-of-function mutation in natriuretic peptide receptor 2 (Npr2) gene is responsible for disproportionate dwarfism in cn/cn mouse

    T Tsuji, T Kunieda

    JOURNAL OF BIOLOGICAL CHEMISTRY   280 ( 14 )   14288 - 14292   2005.4

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  • Evidence for estrogen receptor expression in the bursal epithelial cells of chicks

    Young-Ha Shin, Fumiko Takagi, Sachi Sugita, Song Han, Takehito Tsuji, Asaki Abe, Yasuhiro Kondo

    Animal Science Journal   76 ( 3 )   255 - 259   2005

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    Language：English   Publisher：Blackwell Publishing  

    DOI： 10.1111/j.1740-0929.2005.00264.x

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  • Search for the genes regulating the longitudinal bone growth

    TSUJI Takehito, KIYOSU Chiyo, SOGAWA Chizuru, KUNIEDA Tetsuo

    The journal of animal genetics   32(2) 133-140 ( 2 )   133 - 140   2005

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    Publisher：Japanese Society of Animal Breeding and Genetics  

    DOI： 10.5924/abgri2000.32.2_133

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  • 軟骨形成不全症モデル動物を用いた長管骨の成長を制御する遺伝子の解析

    辻 岳人

    岡山実験動物研究会報   2005

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  • A mutation in the serum and glucocorticoid-inducible kinase-like kinase (Sgkl) gene is associated with defective hair growth in mice

    K Masujin, T Okada, T Tsuji, Y Ishii, K Takano, J Matsuda, A Ogura, T Kunieda

    DNA RESEARCH   11 ( 6 )   371 - 379   2004.12

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  • Expression of the Ellis-van Creveld (Evc) gene in the rat tibial growth plate

    T Tsuji, H Nakamura, A Hirata, T Yamamoto

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   279A ( 2 )   729 - 735   2004.8

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  • New mutant mouse with skeletal deformities caused by mutation in delta like 3 (DII3) gene

    Y Shinkai, T Tsuji, Y Kawamoto, T Kunieda

    EXPERIMENTAL ANIMALS   53 ( 2 )   129 - 136   2004.4

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  • Ellis-van creveld症候群の原因遺伝子であるEvcとLbnの脛骨における発現について

    辻 岳人

    岡山大学農学部学術報告   93   73 - 76   2004

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  • 環境汚染物質の複合曝露が生体に及ぼす影響 －金属結合蛋白質metallothionein誘導を中心として－.

    十川紀夫, 十川千春, 辻 岳人, 山本敏男, 北山滋雄, 小野寺憲治

    応用薬理   2004

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  • Role of osteoclast extracellular signal-regulated kinase (ERK) in cell survival and maintenance of cell polarity

    H Nakamura, A Hirata, T Tsuji, T Yamamoto

    JOURNAL OF BONE AND MINERAL RESEARCH   18 ( 7 )   1198 - 1205   2003.7

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  • Positional cloning of the gene LIMBIN responsible for bovine chondrodysplastic dwarfism

    H Takeda, M Takami, T Oguni, T Tsuji, K Yoneda, H Sato, N Ihara, T Itoh, Kata, SR, Y Mishina, JE Womack, Y Moritomo, Y Sugimoto, T Kunieda

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   99 ( 16 )   10549 - 10554   2002.8

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  • Localization of osteoprotegerin (OPG) on bone surfaces and cement lines in rat tibia

    H Nakamura, T Tsuji, A Hirata, T Yamamoto

    JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY   50 ( 7 )   945 - 953   2002.7

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  • Ultracytochemical study of medullary bone calcification in estrogen injected male Japanese quail

    T Yamamoto, H Nakamura, T Tsuji, A Hirata

    ANATOMICAL RECORD   264 ( 1 )   25 - 31   2001.9

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  • ERK regulates the formation of ruffled borders in osteoclasts.

    H Nakamura, T Tsuji, A Hirata, T Yamamoto

    JOURNAL OF BONE AND MINERAL RESEARCH   16 ( Supp )   S508 - S508   2001.9

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  • The insulin receptor-related receptor (Insrr) gene maps to mouse chromosome 3

    T Tsuji, H Katoh, T Kunieda

    EXPERIMENTAL ANIMALS   50 ( 4 )   359 - 360   2001.7

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  • Immunolocalization of keratan sulfate proteoglycan in rat calvaria

    H Nakamura, A Hirata, T Tsuji, T Yamamoto

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   64 ( 1 )   109 - 118   2001.2

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  • Cloning of the Promoter Region and Expression Analysis of Mouse Insulin Receptor-related Receptor (Irr) Gene

    TSUJI Takehito, SATO Katsunori, KUNIEDA Tetsuo

    Nihon Chikusan Gakkaiho   72 ( 2 )   169 - 171   2001

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    Language：English   Publisher：Japanese Society of Animal Science  

    In the present study, we isolated promoter region of mouse insulin receptor-related receptor (Irr) gene and analyzed its expression in various mouse tissues and embryos in different stages. The nucleotide sequence of the promoter region of the Irr gene showed the presence of putative promoter elements and binding sites for trans-acting factors. The expression analysis revealed the strong expression in kidney, stomach, and embryo at the stage of 14.5 days. These findings were basically consistent with the previously reported findings of rat and human, but we found several differences in the sequence and expression pattern of the gene between mouse and rat or human.

    DOI： 10.2508/chikusan.72.169

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  • 抗エストロジェン剤タモキシフェンはウズラ骨髄骨の吸収を惹起するとともに新たな骨梁形成を誘導した

    平田 あずみ, 中村 浩彰, 辻 岳人, 山本 敏男

    歯科基礎医学会雑誌   42 ( 5 )   422 - 422   2000.8

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  • The down-regulated in adenoma (Dra) gene encoding intestine-specific anion transporter maps to mouse chromosome 12

    T Kunieda, K Takahashi, Y Shinkai, T Tsuji, CW Schweinfest, H Katoh

    EXPERIMENTAL ANIMALS   49 ( 1 )   67 - 68   2000.1

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  • Cloning and Mapping of the Mouse Gpx2 gene Encoding Gastrointestinal Glutathione Peroxidase

    Takehito Tsuji, Yuka Watanabe, Hideki Katoh, Katsunori Sato, Tetsuo Kunieda

    Journal of Veterinary Medical Science   60 ( 5 )   651 - 654   1998

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    Language：English   Publisher：Maruzen Co. Ltd.  

    DOI： 10.1292/jvms.60.651

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  • A Mutation in Endothelin-B Receptor Gene Causes Myenteric Aganglionosis and Coat Color Spotting in Rats

    Tetsuo Kunieda, Taeko Kumagai, Takehito Tsuji, Tsuyoshi Ozaki, Hideaki Karaki, Hiroshi Ikadai

    DNA Research   3 ( 2 )   101 - 105   1996

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    Language：English   Publisher：Universal Academy Press Inc.  

    DOI： 10.1093/dnares/3.2.101

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Presentations

  • 骨成長をもたらす突然変異(dwg)の原因遺伝子の同定

    第３０回日本骨代謝学会  2012 

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  • Mad2l2遺伝突然変異がおよぼすＤＮＡ損傷修復機構への影響

    第３５回日本分子生物学会  2012 

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  • 矮小を呈するstbマウスの組織学的解析および原因遺伝子のマッピング

    第３０回日本骨代謝学会  2012 

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  • sks突然変異マウスのパキテン期精母細胞における核の形態異常

    日本実験動物学会総会  2011 

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  • Roles of Syntaxin2/Epimorphin (Stx2/Epim) in progression of meiosis during mouse spermatogenesis

    2nd World Congress on Reproductive Biology  2011 

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  • The role of Stntaxin2/Epimorphin (Stx2/Epinm) in progression of meisis during dpermatogenesis.

    Society for the study of reproduction 43rd annual meeting  2010 

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  • Cloning of candidate genes for TMEM48 binding proteins, which could be involved in gametogenesis.

    Society for the study of reproduction 43rd annual meeting  2010 

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  • repro34突然変異マウスを用いたStx2/Epim遺伝子の精子形成における機能の解析

    第５７回日本実験動物学会  2010 

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  • マウス精巣においてTMEM48/NDC1と相互作用するタンパク質の探索

    第５７回日本実験動物学会  2010 

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  • Identification of a mutation in the gamma-glutamyl transferase 1 (Ggt1) gene causing dwarfism and cataracts in dwg/dwg mice

    23nd International Mammalian Genome Conference  2009 

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  • Stx2/Epim遺伝子の突然変異により精子形成に異常を呈するrepro34マウスの解析

    第１０２回日本繁殖生物学会  2009 

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  • 見島牛における和牛の諸形質に関与する遺伝子変異の解析

    本動物遺伝育種学会第１０回記念大会  2009 

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  • DWARFISM AND CATARACTS IN MICE WITH dwg LOCUS ARE CAUSED BY THE MUTATUION IN Ggt1 GENE

    Mouse Genetics & Genomics:Development & Disease.  2009 

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  • 骨成長不全および白内障をもたらす突然変異(dwg)の原因遺伝子の同定

    第５６回日本実験動物学会総会  2009 

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  • 減数分裂第一分裂前期で精子形成が停止するENU 誘発突然変異(repro23 )マウスの解析

    第５５回日本実験動物学会総会  2008 

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  • 排卵過程におけるエンドセリンファミリーの役割について

    第９回日本動物遺伝育種学会  2008 

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  • 精子形成が減数分裂第一分裂前期で停止するＥＮＵ誘発突然変異repro23マウスの解析

    第９回日本動物遺伝育種学会  2008 

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  • 精子形成に特異的なSpata4遺伝子の機能の解析

    第９回日本動物遺伝育種学会  2008 

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  • Hypomorphic mutation in mouse Nppc gene causes retarded bone growth due to impaired endochondral ossification

    22nd International Mammalian Genome Conference  2008 

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  • Npr2変異マウスにおける脂肪重量減少についての解析

    第５５回日本実験動物学会総会  2008 

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  • Limbinのノックアウトマウスは軟骨低形成、短肢症を呈する〜Ellis-van Creveld syndromeの疾患モデルマウスの作製〜

    第２０回日本軟骨代謝学会  2007 

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  • FKBP6欠損ラット(TTラット)の精母細胞に出現するリボソーム集塊の原因解析

    第100回日本繁殖生物学会大会  2007 

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  • 生殖生命科学研究のためのモデル動物の確立

    第２４回日本疾患モデル学会総会  2007 

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  • Limbinノックアウトマウスは軟骨内骨化の異常による骨成長遅延を呈する

    第２５回日本骨代謝学会  2007 

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  • Thtマウスにおける初期胚の発生停止と染色体逆位

    第５４回日本実験動物学会  2007 

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  • FKBP6欠損ラット(TTラット)の精母細胞に出現するリボゾーム集塊の原因解析

    第５４回日本実験動物学会  2007 

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  • Linkage mapping of locus responsible for Forelimb-girdle Muscular Anomaly of japanse black cattle on BTA26

    第107回日本畜産学会  2007 

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  • ミュータントマウスを用いた卵胞形成におけるCNP/NPRB の機能の解析

    第２４回日本疾患モデル学会総会  2007 

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  • CNP/NPRB plays an important roles in follicular development and oocyte maturation.

    Society for the study of reproduction 40Th annual meeting  2007 

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  • A mutation causing abnormal splicing of Tmem48/NdC1 gene is responsible for impaired gametogenesis in SKS mutant mouse.

    Society for the study of reproduction 40Th annual meeting  2007 

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  • Ehマウスが呈する口蓋裂の発生機構および原因遺伝子の解析

    第２３回日本疾患モデル動物学会  2006 

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  • 骨格異常を呈するThtマウスの連鎖解析および表現型の解析

    第５３回日本実験動物学会  2006 

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  • 骨格および配偶子形成に異常を呈するミュータントマウスにおける新規膜貫通タンパク質遺伝子の突然変異

    第５３回日本実験動物学会  2006 

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  • Edmrb遺伝子の挿入レトロトランスポゾンの欠失により生じたpiebald復帰突然変異体

    第５３回日本実験動物学会  2006 

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  • マウスjaggedtail(jg)突然変異の染色体マッピングおよび形態学的解析

    第２３回日本疾患モデル動物学会  2006 

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  • Comparative mapping of bovine chromosome 18 revealed candidate genes for bovine multiple ocular defects

    第１０４回日本畜産学会  2005 

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  • Cloning of bovine F8 gene and a mutation associated with factor VIII deficiency of Japanese brown cattle

    第１０４回日本畜産学会  2005 

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  • piebald 復帰突然変異体におけるエンドセリンBレセプター遺伝子の解析

    第２２回日本疾患モデル学会  2005 

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  • アロキサン誘発糖尿病感受性とIdd10遺伝子座との関連について

    第２２回日本疾患モデル学会  2005 

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  • SLWマウスにおけるCNPレセプター(NPRB)欠損が消化管に与える影響

    第２２回日本疾患モデル学会  2005 

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  • NPRB変異マウス(cn/cn)の雌における生殖機能の解析

    第２２回日本疾患モデル学会  2005 

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  • 軟骨形成不全症を呈する突然変異(cn/cn)マウスの原因遺伝子の同定

    第２８回日本分子生物学会  2005 

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  • Natriuretic peptide receptor 2 遺伝子におけるミスセンス変異は長管骨の成長障害をもたらす

    第２３回日本骨代謝学会  2005 

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  • 矮小を呈するSLWマウスにおける原因遺伝子の同定

    第５２回日本実験動物学会  2005 

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  • A loss-of-function mutation in natriuretic peptide receptor 2 (Npr2) gene is responsible for mouse achondroplasia.

    第１８回マウスゲノム会議  2004 

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  • Linkage mapping of the locus responsible for bovine congenital eye anomaly on the bovine choromosome 18.

    第２９回 国際動物遺伝学会議  2004 

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  • Cloning of bovine F11 gene and identification of a mutation responsible for factor XI deficiency of Japanese black cattle

    第２９回 国際動物遺伝学会議  2004 

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  • 新規矮小マウス(SLW)における形態学的解析と原因遺伝子のマッピング

    第５１回日本実験動物学会総会  2004 

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  • 被毛異常ミュータントマウスの原因となるSgkl遺伝子における突然変異

    第２１回日本疾患モデル動物学会  2004 

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  • A mutation in the serum and glucocorticoid-inducible kinase-like kinase (Sgkl) gene is responsible for defective hair growth in mice.

    第１８回マウスゲノム会議  2004 

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  • An insertion of a retrotransposon in the Ednrb gene cause piebald coat color of the s/s mouse.

    第１８回マウスゲノム会議  2004 

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  • 毛色異常突然変異piebald(s)におけるEdnrb遺伝子へのレトロトランスポゾンの挿入

    第５１回日本実験動物学会総会  2004 

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  • 黒毛和種における血液凝固第XI因子欠乏症の遺伝子診断法の確立

    第１０３回日本畜産学会  2004 

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  • Identification of the breakpoints of choromosomal inversions in Hairy Ear (EH) and Koala (KOA) mutant mice.

    国際マウスゲノム会議  2003 

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  • Eh(Hairy ear)マウスにおける染色体逆位の切断点の決定

    第５０回日本実験動物学会  2003 

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  • 雄性仮性半陰陽ラット(mp/mp)における原因遺伝子の同定

    第２０回日本疾患モデル学会  2003 

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  • 環境汚染物質の複合曝露が生体に及ぼす影響 －金属結合蛋白質metallothionein 誘導を中心として－

    第５回応用薬理シンポジウム  2003 

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  • 破骨細胞におけるMAPkinase系ERKに関する形態学的解析

    第107回日本解剖学会  2002 

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  • 四肢短縮小人症Ellis-van Creveled症候群の原因遺伝子EVCとIHHの成長板軟骨におけ る発現

    第25回日本分子生物学会  2002 

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  • エナメル芽細胞におけるExtracellular signal-regulated kinase (ERK) に関する形 態学的研究

    第44回歯科基礎医学会学術大会  2002 

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  • Positional cloning of a novel gene, LIMBIN, responsible for a bovine chondrodysplastic dwarfism.

    第28回 国際動物遺伝学会議  2002 

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  • 破骨細胞のExtracellular Signal Regulated kinase (ERK)に関する形態学的解析

    第20回骨代謝学会  2002 

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  • Positional cloning of a novel gene, LIMBIN, responsible for a bovine chondrodysplastic dwarfism.

    第２５回分子生物学会  2002 

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  • 四肢短縮小人症Ellis-van Cleveled症候群の遺伝子であるEVC遺伝子の軟骨細胞にお ける発現

    第20回骨代謝学会  2002 

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  • 吸収骨表面、セメントラインにおけるオステオプロテジェリン局在

    第43回歯科基礎医学会  2001 

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  • MAP kinase系ERKは破骨細胞の波状縁形成を制御する

    第19回日本骨代謝学会  2001 

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  • 鳥類の骨髄骨におけるオステオポンチンの免疫組織化学的研究

    第106回日本解剖学会  2001 

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  • 骨組織におけるオステオプロテジェリン局在

    第106回日本解剖学会総会  2001 

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  • ウズラ骨髄骨基質のケラタン硫酸と石灰化

    第１０５回日本解剖学会  2000 

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  • 骨髄骨石灰化におけるケラタン硫酸とsulferの分布について

    第４２回歯科基礎医学会  2000 

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Research Projects

  • 動物集団における近交退化の分子メカニズムの解明

    Grant number：20H00446  2020.04 - 2024.03

    日本学術振興会  科学研究費助成事業  基盤研究(A)

    国枝 哲夫, 大月 純子, 佐々木 慎二, 桃沢 幸秀, 藤原 靖浩, 揖斐 隆之, 辻 岳人, 渋谷 周作

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    Grant amount：\44720000 （ Direct expense: \34400000 、 Indirect expense：\10320000 ）

    野間馬の全個体について家系情報を収集し、それらに基づいて各個体の近交係数を調べた結果、野間馬集団の過去46年間の平均近交係数は0.195であり、また平均近交係数は1986年以降漸次上昇していることが明らかになり、近交化が進行していることが確認された。また、野間馬集団中の多くの個体のDNAを用いて、DNAマイクロアレイにより全ゲノムを網羅した約7万の一塩基多型（SNPs）のタイピングを行うと共に、主要組織適合性遺伝子複合体（MHC）のハプロタイプのタイピングを行った。その結果、SNPs のタイピング結果より求めた野間馬集団の平均近交係数は0.15であり、家系情報より求めた平均近交係数より若干低いもののやはり高い傾向を示した。さらにMHCクラスII領域の10遺伝子座のタイピングを行ったところ、野間馬の集団中には4ハプロタイプしか存在しないことが明らかになった。これらの結果から、当初の予想通り、野間馬の集団の近交化はかなり進行していることが確認された。また、現在飼育されている野間馬の全個体について、臨床診断、血液検査等により異常の詳細な調査も実施している。
    これまでに世界各国から収集したウマのDNAサンプルを用いて、PRDM9遺伝子の塩基配列を解析した結果、これら動物種においても他の動物種と同様に極めて多様性が高いことが確認された。またヒトにおいても日本人の集団中においてPRDM9遺伝子のZinc-Finger Domainの多様性は高く多くのハプロタイプが存在することが確認された。
    さらに、黒毛和種の集団で発生が報告された、特徴的な症状を呈して生後間もなく死亡する疾患の発症個体について、家系の調査と共に広く全ゲノムの塩基配列の解析を行ったところ、特定の遺伝子を含むゲノム広い領域における欠失が確認され、近交化にともなうゲノムの不安定化がその原因である可能性も示唆された。

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  • 不妊マウスから単離された新規原因遺伝子Adamtsl2による卵巣機能制御の解明

    Grant number：20K06470  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)

    辻 岳人, 若井 拓哉

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

    正常マウス卵巣におけるADAMTSL2タンパク局在を明らかにするため、４領域の異なる組換えタンパクを抗原とするウサギポリクローナル抗体を作製し、Adamtsl2遺伝子変異をもつミュータントマウスと正常マウスの卵巣およびその他組織における免疫染色を実施した。それぞれの組織において一定の陽性反応は得られるものの特異性が十分と判断できるシグナルを得ることかができなかった。さらに、in situハイブリダイゼーションによる卵巣におけるAdamtsl2遺伝の発現様式についても解析を行なった。結果として、シグナル強度は弱いものの特定の細胞に限定した発現様式を確認した。また、Adamtsl2遺伝子変異による卵巣、子宮、下垂体への影響を解析するため、RNA-seqによるミュータントマウスと正常マウスでの網羅的な遺伝子発現量の比較をおこなった。その結果、ミュータントマウスにおいてステロイドホルモン合成経路の酵素や複数のホルモンの遺伝子発現が際立って低下していることが明らかになった。一方、これまでに相互作用因子として報告されている細胞外マトリックスの遺伝子発現には差がなかった。さらに、Adamtsl2遺伝子変異によるタンパク局在への影響を解析するため、前年度作製した全長のAdamtsl2遺伝子発現ベクターおよび遺伝子変異から予想される短縮型のAdamtsl2遺伝子発現ベクターを培養細胞へ導入してタンパク局在への変化を比較した。その結果、正常な全長のADAMTSL2タンパクは細胞外での局在が確認されたが、短縮型では細胞内にのみ局在が確認され、異常タンパクとして小胞体へ取り込まれていることが予想された。

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  • Roles of CNP signaling pathway in follicular development and atresia

    Grant number：25450395  2013.04 - 2017.03

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

    tsuji takehito, YASODA akihiro, KUNIEDA Tetsuo

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    Grant amount：\5330000 （ Direct expense: \4100000 、 Indirect expense：\1230000 ）

    In this study, we investigated whether C-type natriuretic peptide (CNP) functions to regulate follicular development and atresia. The numbers of ovarian follicles in mice injected with CNP and CNP transgenic mice showed no difference compared with those of normal mice. We showed the expression of Npr3 gene in granulosa cells of ovarian follicles, but no alterations were observed in the ovary of mice with a mutation in Npr3 gene. Next, we investigated CNP function in cultured early antral follicles, and found that a moderate inhibitory effect of CNP on apoptosis in granulosa cells. We conclude that CNP may have the potential to control follicular atresia via inhibition of apoptosis in granulosa cells.

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  • Investigation for effects of blood coagulation factor XI deficiency on productivity to the genetic improvement of Japanese beef cattle

    Grant number：23380166  2011.04 - 2014.03

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

    KUNIEDA Tetsuo, TSUJI Takehito, IBI Takayuki, FUNAHASHI Hiroaki, ACOSTA Tomas

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    Grant amount：\18200000 （ Direct expense: \14000000 、 Indirect expense：\4200000 ）

    Blood coagulation factor XI deficiency is a hereditary disorder observed in Japanese Black cattle. While the clinical condition of this disorder is mild, the economical impact of this disorder is apprehended to be serious because of the remarkably high allelic frequency of the mutant allele in the population of Japanese Black cattle. We, therefore, investigated the effect of the factor XI deficiency on defects in pregnancy and delivery and carcass traits using large samples of Japanese Black cattle. As a result, we found that affected animals of factor XI deficiency have a tendencies for increased frequencies of abortion and stillborn, but no effect on the condition of calf and carcass traits. These findings will be useful to future breeding of Japanese Black cattle.

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  • Study on the role of CNP in the regulation of oocyte meiosis

    Grant number：22580332  2010 - 2012

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

    TSUJI Takehito, KUNIEDA Tetsuo

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

    This study investigated the role of CNP/NPR2 signaling in the regulation of oocyte meiosis, and obtained novel findings as follows: (1) CNP and its receptor (NPR2) were expressed in ovarian granulosa cells (2) mice with a mutation in Npr2 gene showed precocious resumption of oocyte meiosis in antral follicles (3) CNP was downregulated in response to LH signaling. These results suggest that the arrest of oocyte meiosis is maintained by CNP/NPR2 signaling from the granulosa cells that is relieved by LH signaling.

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  • Heparanase localization and stimulatory effect during formation and regeneration of periodontal tissues

    Grant number：21592571  2009 - 2011

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

    HIRATA Azumi, UENO Takaaki, TSUJI Takehito, MATSUMURA Tatsushi, SUGAHARA Toshio

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    Grant amount：\4550000 （ Direct expense: \3500000 、 Indirect expense：\1050000 ）

    We have previously reported that heparanase may contribute to root formation, especially cementoblast differentiation, by degrading perlecan in the dental basement membrane. The aim of this study was to determine whether heparanase might function in periodontal development and regeneration. Our studies indicated that 1) Heparanase localization was observed in osteoblasts and in the basal cells of the oral epithelium. 2) Palatal epithelial cells were labeled with intense heparanase signal during palatogenesis ; in contrast, perlecan labeling was sparse in the basement membrane of medial epithelial seam. 3) Exogenous heparanase might exhibit proliferative potential to dental mesenchymal stem cells in the apical space. These findings suggested that heparanase may have an important role during the process of formation and regeneration of periodontal tissues.

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  • Translational research for novel therapeutic strategy for skeletal dysplasias

    Grant number：21591176  2009 - 2011

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

    YASODA Akihiro, TSUJI Takeito

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    Grant amount：\4550000 （ Direct expense: \3500000 、 Indirect expense：\1050000 ）

    We performed the translational research of the CNP/GC-B system for impaired skeletal growth of skeletal dysplasias using mice model of skeletal dysplasias. We studied interactions between the CNP/GC-B system and molecules that play essential roles on endochondral bone growth using cell lines and organ culture system. Further we investigated the effects of the CNP GC-B system on impaired bone growth in mice models of skeletal dysplasias.

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  • Genetic investigation of the genes responsible for hereditary disorders of Japanese Black cattle and its functional analysis

    Grant number：20248029  2008 - 2010

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

    KUNIEDA Tetsuo, TSUJI Takehito, UCHIDA Kazuyuki, MORITOMO Yasuo, YAMATO Osamu, WATANABE Daisaku

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    Grant amount：\47710000 （ Direct expense: \36700000 、 Indirect expense：\11010000 ）

    We have attempted to identify the gene responsible for hereditary disorders of Japanese Black cattle and develop the genetic diagnosis systems to detect the mutations. Consequently, we have revealed that a nonsense mutation of the GFRA1 gene is responsible for forelimb-girdle muscular anomaly and developed the genetic diagnosis systems for this disorder by PCR-RFLP method. For other disorders including short mandible and renal agenesis, we have collected samples of affected animals and performed linkage analysis using these samples. As a result, the locus for short mandible and renal agenesis was mapped on a particular region of bovine chromosome. These finding can be used to develop genetic diagnosis system for this disorder.

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  • The role of C-type natriuretic peptide in fat accumulation

    Grant number：19780217  2007 - 2008

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

    TSUJI Takehito

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    Grant amount：\3060000 （ Direct expense: \2700000 、 Indirect expense：\360000 ）

    これまで骨の成長に重要であることが知られていたC 型ナトリウム利尿ペプチド(CNP)が、新たな機能として体内の脂肪蓄積の量を決定することに関与する可能性が明らかになった。その作用は、脂肪細胞の分化を制御する遺伝子に作用することによる。今回の研究の成果は、CNPが脂肪の形成に関わる制御機構の解明に向けた新たな突破口となることが考えられる。

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  • Molecular cloning and functional analysis of gene responsible for multiple ocular defect of cattle

    Grant number：18380167  2006 - 2007

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

    KUNIEDA Tetsuo, TSUJI Takehito, UCHIDA Kazuyuki

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    Grant amount：\17030000 （ Direct expense: \15500000 、 Indirect expense：\1530000 ）

    Multiple ocular defects (MOD) in cattle is an autosomal recessive hereditary disorder characterized by dysplasia of the lens, retinal detachment, persistence of the hyaloid artery, and microphthalmia. The locus responsible for MOD was mapped to the proximal region of bovine chromosome 18. We further refined the localization of the MOD locus to within a 1.1 Mb interval on bovine chromosome 18 by haplotype analysis and determined the nucleotide sequences of all genes in this region. A comparison of the nucleotide sequences of these genes between affected and normal animals revealed a one-nucleotide insertion in exon 2 of the WFDCl gene, which resulted in a frame shift mutation. WFDCl is a small secretory protein containing a WAP-type four disulfide core domain. Specific expression of Wfdcl was observed in the lens, retina, and optic nerves of embryonic and adult mouse eyes by immunohistochemical staining and in situ hybridization. Although the exact function of WFDCl in mammalian development remains unknown, the present findings showing that this mutation in WFDCl resulted in multiple eye defects and that the gene is specifically expressed in embryonic and adult eyes demonstrated the essential role of WFDCl in mammalian eye development. This is the first report providing direct evidence for the involvement of WFDCl in a particular process of mammalian development.

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  • 生殖ジェノミックスのための研究基盤としてのモデル動物の確立

    Grant number：18650112  2006

    日本学術振興会  科学研究費助成事業  萌芽研究

    国枝 哲夫, 辻 岳人, 松田 潤一郎

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    Grant amount：\3300000 （ Direct expense: \3300000 ）

    Fkbp6は配偶子形成に重要な役割を担うと考えられている遺伝子であるが、その詳細な機能は不明である。そこで、本研究では酵母ツーハイブリッド法によりFKBP6と相互作用するタンパク質の同定を試みた。その結果、特定の翻訳開始因子の遺伝子、リボゾーム大サブユニットを構成するタンパク質の遺伝子、機能は不明であるが、精巣特異的に発現する遺伝子が同定された。次にこれらの遺伝子が実際に精子形成に関与しているかどうかを調べるために、ENU誘発突然変異マウスのスクリーニングにより、これらの遺伝子に変異を持つマウスを得ることを試みた。すなわち数千個体のENU誘発突然変異マウスのゲノムDNAについて、これらの遺伝子の塩基配列を解析することで、当該遺伝子に変異を持つ個体を特定した。これらの変異を持つマウスは配偶子形成を中心とする生殖機能の解析のための優れたモデル動物となることが期待される。
    また、米国のジャクソン研究所においては同様にENUの投与により誘発されたランダムな突然変異を持つマウスから、生殖機能に異常を持つミュータントマウスを多数樹立している。これらのミュータントマウスの原因遺伝子はまだ不明であることから、これらのマウスの原因遺伝子を同定することで、生殖機能に関わる新たな遺伝子を同定することが可能であると考えられる。そこで、精子形成に異常を呈するミュータントマウスおよび生殖細胞が欠損するミュータントマウスをジャクソン研究所より導入し、その原因遺伝子の染色体マッピングと詳細な表現型の解析を行った。その結果、これらのマウスのうちの1系統に、精子形成異常の原因と考えられる遺伝子の変異が存在していることが明らかとなった。これらのマウスも生殖機能の解析のための優れたモデル動物となることが期待される。

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  • 減数分裂に異常を呈するFKbp6欠損動物を用いた配偶子形成の分子機構の解明

    Grant number：16045211  2004 - 2005

    日本学術振興会  科学研究費助成事業  特定領域研究

    国枝 哲夫, 辻 岳人, 野口 純子

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    Grant amount：\5400000 （ Direct expense: \5400000 ）

    精子形成不全を呈するas/asラットでは、雌では卵形成は正常に進行し生殖可能であるが、雄では減数分裂に異常を呈し、精子形成は減数分裂のパキテン期で停止する。これまでに研究によりFkbp6遺伝子にas/asラットに特異的な突然変異を同定し、本遺伝子の機能の欠損が精子形成不全の原因であることを明らかにしている。さらにFKBP6は相同染色体の対合装置であるシナプトネマ構造を構成するととともに、性染色体の対合にも深く関与することが明らかとなっている。以上のことから、FKBP6は減数分裂時の相同染色体の対合、特にほ乳類の雄に特異的なXおよびY染色体の正常な対合と分離に重要な役割を果たしていると考えられ、FKBP6の減数分裂における機能を解析することで、減数分裂の進行に必須である染色体の対合と分離の分子機構が解明されることが期待される。
    これまでに相同染色体の対合異常により減数分裂が停止する多くのミュータントが酵母かマウス等で報告されているが、それらの中には正常な組換えの進行に必要とされるDNA二本鎖切断修復やミスマッチ修復に関するタンパク質は、正常な組換え頻度の調節に必要とされるタンパク質、あるいはヒートショックタンパク質等が知られている。したがってFkbp6もこれらの過程に関与している可能性が考えられ、本研究ではFkbp6の減数分裂における機能を明らかにすることを目的として、Fkbp6^<-/->Fkbp6^<+/->Fkbp6^<+/+>マウスの間でDNA二本鎖切断修復機能、減数分裂の進行に対する高温曝露の影響および組換え頻度と干渉現象について比較した。その結果、いずれにおいてもFkbp6^<-/->Fkbp6^<+/->Fkbp6^<+/+>マウスの間で有意な違いは認められず、FKBP6はこれらの機能に関与していないことが結論づけられた。

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  • Cloning of causative gene and developing the DNA diagnosis system for Japanese beef cattle

    Grant number：16380188  2004 - 2005

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

    KUNIEDA Tetsuo, TSUJI Takehiro, OGAWA Hiroyuki

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    Grant amount：\15300000 （ Direct expense: \15300000 ）

    The incidence of various hereditary diseases has been reported in Japanese beef cattle and these diseases have caused serious problems for breeding and raising beef cattle. In this study, we attempted to identify causative genes and to develop DNA diagnosis systems for the following two hereditary diseases in Japanese beef cattle. 1)Multiple ocular defects is hereditary ocular disorder showing developmental defects of the lens, retina and iris, persistent embryonic eye vascularization, and microphthalmia. We mapped the locus for this disease to bovine chromosome 18 by linkage analysis. Then, we cloned and sequenced several potential candidate genes for the disorder including the MAF and FOXC2 genes, but no causative mutation for the disease was identified. However, we established the marker-assisted selection for identification of carriers of the disease by using linked markers. 2)Hemophilia A is a congenital severe bleeding disorder characterized by subcutaneous hematoma and hemorrhage into muscles resulting from a deficiency of blood coagulation factor VIII. Recently two cases of Hemophilia A were reported in Japanese brown cattle. In this study we identified a nucleotide substitution in factor VIII gene resulting in an amino acid substitution of Leusin to Histidin as a possible cause of the deficiency. We also developed a simple and effective PCR-based diagnostic method to identify carriers of this disorder, using a mismatch primer in combination with a restriction enzyme digestion. The mapping and identification of the genes responsible for these diseases provided the basis for DNA-based diagnostic systems for these diseases.

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  • 軟骨形成不全症の原因遺伝子(Evc)による軟骨細胞の増殖・分化制御機構の解析

    Grant number：14771002  2002 - 2003

    日本学術振興会  科学研究費助成事業  若手研究(B)

    辻 岳人

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    Grant amount：\2900000 （ Direct expense: \2900000 ）

    昨年度の研究によりEvc遺伝子の前肥大軟骨細胞における強い発現が確認され、Evc遺伝子が軟骨細胞の分化に関与することが推測された。本年度においては、さらに軟骨細胞における機能的役割を明らかにすることを目的として解析をおこなった。まず、未分化軟骨細胞株であるATDC5にEvc遺伝子を安定的に過剰発現させる細胞を樹立し、その細胞の変化について解析することを試みた。単離した大半の細胞は増殖活性が非常に悪く、解析するまでの細胞数には至らなかった。最終的に過剰な発現が認められたのは1系統のみであった。通常、分化誘導培地でATDC5を培養しなければ軟骨細胞へ分化しないが、この細胞系統は非分化誘導培地にもかかわらず細胞凝集を形成しながら増殖した。また細胞の形態も繊維芽細胞様から丸みを帯びた軟骨細胞様に変化していた。これらの結果から、Evc遺伝子の過剰発現により軟骨細胞への分化が誘導されている可能性が考えられた。
    近年、Ellis-van Creveld症候群の原因遺伝子として本研究で取り上げたEvc遺伝子だけでなく、新たにLim-bin(Lbn)遺伝子が原因遺伝子であることが明らかにされた。さらに、両遺伝子は染色体上の位置が近接していることから、機能的になんらかの関連性が推測されることから、各組織における発現パターンについて解析を行った。各週齢のラット脛骨骨端および4週齢の肝臓、脳、肺、心臓、腎臓、骨格筋について半定量的RT-PCRによりEvcとLbn遺伝子の発現パターンを解析した。その結果、両遺伝子とも、すべての組織で類似した発現パターンを示した。これらの結果は、両遺伝子の関連性をさらに示唆する結果であり、軟骨細胞においても機能的な関連性があるものと考えられた。
    本研究により、Evc遺伝子が軟骨細胞の分化誘導に関与すること、またその作用はLbn遺伝子と関連性が推測され、軟骨細胞分化の制御機構でのEvc遺伝子の重要性が初めて示された。

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  • Analysis of the worse effects of Bisphenol A on the hepatic toxicity by the environmental pollutant

    Grant number：13671936  2001 - 2003

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

    ONODERA Kenji, SOGAWA Norio, YAMAMNOTO Toshio, KITAYAMA Shigeo, TSUJI Takehito, SOGAWA Chiharu

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    Grant amount：\1400000 （ Direct expense: \1400000 ）

    To clarify the toxic action of the environmental pollutants on the living body is one of the mist important problems. By the way, the facts which the several environmental pollutants have been detected in human umbilical cords collected from healthy newborns indicate that we are not exposed to a single chemical compound. However, there are few reports which investigated the effects of environmental pollutants on the living body from the point of view of the composite exposure. We have been reported that there are gender differences in the synthesis of hepatic metallothionein (MT) which is considered to play an important role in the detoxification of cadmium (Cd), and demonstrated that sex hormones related to these gender differences. It is considered that these findings are very interesting results suggesting the possibility of the worse effects of female sex hormon on the Cd-induced liver damage. Therefore, we investigated the effects of Bisphenol A (BPA), an estrogen-like endocrine disruptor, on the hepatic MT synthesis and damage after Cd injection in order to clarify the effects of the composite exposure of environmental pollutants on the living body.
    As the results, BPA (100μg/kg) reduced the Cd (0.5μg/kg)-induced MT mRNA expression and MT contents in the liver. And neither BPA alone nor Cd could increase GPT and GOT activities, however, both GPT and GOT activities of Cd-injected mice with pretreatment of BPA (100μg/kg) were increased to compare with the vehicle-pretreated mice. Moreover, under the conditions without MT synthesis, female MT-null mice were more tolerant than male MT-null mice against the Cd-induced toxicity. These results indicate that Cd and BPA showed the synergistic action to the liver damage, anal the possibility of the mechanisms of defense against the acute Cd-induced toxicity are different between male and female.

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  • エナメル質形成不全の遺伝子発現異常によるエナメル芽細胞の分化制御機構の解析

    Grant number：12771083  2000 - 2001

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    辻 岳人

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

    前年度おこなったエナメル芽細胞を用いたサブトラクションでは、使用した歯胚のRNAにおけるエナメル芽細胞の割合が低く、そのためにエナメル質形成不全症ラットと正常ラットのエナメル芽細胞で発現量の差を示す遺伝子を効率よく単離することが出来なかった。本年度においては、エナメル芽細胞を含む上皮由来のエナメル器のみを材料に用いることにより、エナメル芽細胞で特異的に発現する遺伝子の同定をさらに試みた。まず、生後3日目のエナメル質形成不全ラットと正常ラットから単離した歯胚にトリプシン/コラゲナーゼ処理を行うことで歯胚のエナメル器と歯乳頭を分離しエナメル器からRNAを抽出した。昨年度と同様にサブトラクション法により作製したライブラリーから、コロニーアレー法による二次スクリーニングを行い、エナメル質形成不全ラットにおいて発現量の少ない遺伝子を単離した。これらの遺伝子には、アメロゲニンだけでなくエナメリン、アメロブラスチンなどのエナメルタンパクが多く含まれていた。このことは前年度における結果ではアメロゲンニンが単離されたのみであったことと比較して、本年度に使用した方法が有効であることを示している。既知のエナメルタンパク以外の遺伝子としては、8個の遺伝子を単離した。これらの遺伝子は塩基配列から相向性を示す遺伝子をBlastで検索したが、すべてこれまでに単離されていない遺伝子であった。これらの遺伝子について発現の特異性を確認するために、歯胚を含む各臓器からのRNAを用いたRT-PCRを行った。その結果、2個の遺伝子について歯胚に特異的に発現している傾向を示したことから、これらの遺伝子がエナメル質形成不全ラットの原因遺伝子となんらかの関連性があると考えられた。これらの研究結果は、エナメル芽細胞の分化を制御する機構を解明するために重要な手がかりとなると思われる。

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