配偶子分化の分子機構

1991 Fiscal Year Annual Research Report

• Project/Area Number: 03207101
• Research Institution: The Open University of Japan
• Principal Investigator: 毛利 秀雄 放送大学, 教養学部, 教授 (70012268)
• Co-Investigator(Kenkyū-buntansha): 高橋 迪雄 東京大学, 農学部, 教授 (30011943) ; 岸本 健雄 東京工業大学, 生命理工学部, 教授 (00124222) ; 大島 博幸 東京医科歯科大学, 医学部, 教授 (60013934) ; 石居 進 早稲田大学, 教育学部, 教授 (90063528) ; 安部 真一 熊本大学, 理学部, 教授 (90109637)
• Keywords: 精子形成 / 卵成熟 / セルトリ細胞 / X精子とY精子 / サイクリン / SRY遺伝子 / 雄性不妊 / 生殖腺刺激ホルモン

Research Abstract

今年度における本研究班の研究実績は次の通りである。
1.X,Y精子の分離については、ヒトおよびマウスで無担体電気泳動を行なった各分画をPCR法にかけ、両者の分離が確かなことを示した。
2.イモリ精細胞の核伸長の機構に関連して、尾鞘の微小管束より微小管結合タンパク質を抽出し、微小管との相互作用を明らかにした。
3.ラット精巣におけるセルトリ細胞の細胞間コミュニケ-ションは軽度で、造精過程の同調性は精原細胞間の同調性によるものと考えられる。
4.哺乳類精子の成熟機構に関し、精巣上体分泌液による精子表層の修飾やイオン輸送活性の変化を解析し、アデニルシクラ-ゼを精製した。
5.Sl,W遺伝子がマウス精原細胞の増殖と分化に重要な役割を果すことを明らかにした。セルトリ細胞は活性をもつSl遺伝子産物を産生する。
6.XX男性では外陰部が正常な例でSRY遺伝子の転座を認めたが、尿道下裂の例ではなかった。真性半陰陽と多睾丸でSRYを検出した。
7.亜種間雑種の雄性不妊F1と正常間キメラを解析し、不妊精巣では生殖細胞の接合期において細芯分化が阻害されていることを明かにした。
8.ヒトデ卵を用いて減数分裂周期制御の特異性をサイクリンについて検討し、第一ではBタイプ、第二ではA、B両タイプの関与を認めた。
9.単一卵・胚の成熟過程の生化学的分析法として、microimmunoblottingおよびmicroslab gel電気泳動法を開発し、酵素等の変動を見た。
10.リボヌクレア-ゼと部分的相同な分子量35KDの卵成熟抑制因子を分離し、また卵母細胞の退行抑制作用をもつ因子の作用を明らかにした。
11.ウズラの黄体形成ホルモンのαとβサブユニットのcDNAのクロ-ニングを行い、αに対してβは変化に富んだものであることを示した。
12.アクチビンがゴナドトロ-フの分化調節因子であること、およびプロラクチンの作用がTGFーβで仲介されることを明らかにした。

Research Products

• [Publications] Tsujimura,T.,Hirota,S.,Nomura,S.,Niwa,Y.,Yamazaki,M.,Tono,T.,Morii,E.,Kim,H.,Kondo,K.,Nishimune,Y.,Kitamura,Y.: "Characterization of Ws mutant allele of rats:A 12ーbase deletion in tyrosine kinase domain of cーkit gene." Blood. 78. 1942-1946 (1991)
• [Publications] Koshimizu,U.,Sawada,K.,Tajima,Y.,Watanabe,D.,Nishiune,Y.: "Whiteーspotting mutations affect the regenerative differentiation of testicular germ cells:Demonstration by experimental cryptorchidism and its surgical reversal." Biology of Reproduction. 45. 642-648 (1991)
• [Publications] MaeKawa,M.,Nishimune,Y.: "Inーvitro proliferation of germ cells and supporting cells in the neonatanl mouse testis." Cell and Tissue Research. 265. 551-554 (1991)
• [Publications] Sawada,K.,Sakamaki,K.,Nishimune,Y.: "Effect of the W mutation,for white belly spot,on testicular germ cell differentiation in mice." Journal of Reproduction and Fertility. (1992)
• [Publications] Tajima,Y.,Onoue,H.,Kitamura,Y.,Nishimune,Y.: "Biologically active kit ligand growth factor is produced by mouse Sertoli cells and is defective in Sl^d mutant mice." Development. 113. 1031-1035 (1991)
• [Publications] Koshimizu,U.,Watanabe,D.,Tajima,Y.,Nishimune,Y.: "Effects of W (cーkit) gene mutation on gemetogenesis in male mice.Inborn loss of germ cells causes the partial agametic segments of seminiferous tubules in W^f /W^ftestis." Development. (1992)
• [Publications] Matsuyama,S.,Shiota,K.,Nishihara,M.,Takahashi,M.: "Macrophages in the corpus luteum mediate luteotropic action of prolactin by producing transforminq,growtr.facforーβ." Proceeding of Japanese Society of Reproductive Immunology. (1992)
• [Publications] Saito,S.,Matsuyama,S.,Yamanouchi,K.,Takahashi,M.: "Changes in ratios of phagocytotic macrophages in the splenic adherent cells during esterous cycle and pseudopregnancy in rat." Journal of Reproduction and Development. 38. (1992)
• [Publications] Yamanouchi,K.Matsuyama,S.Nishihara,M.,Shiota,K.,Tachi,C.Takahashi,M.: "Splenic macrophages enhance prolactinーinduced progestin secretion from mature rat granulosa cells in vitro." Biology of Reproduction. 46. (1992)
• [Publications] Nakagome,Y.et al.: "PCR detection of distal Yp sequences in an XX true hermaphrodite." American Journal of Medical Genetics. 41. 112-114 (1991)
• [Publications] Tajima,Y.,Sakamaki,K.,Watanabe,D.,Koshimizu,U.,Matsuzawa,T.,Nishimune,Y.: "SteelーDickie (sl^d) mutation affects both maintenance and differentiation of testicular germ cells in mice." Journal of Reproduction and Fertility. 91. 441-449 (1991)
• [Publications] Niwa,Y.,Kasuga,T.,Ohno,K.,Morimoto,M.,Yamazaki,M.,Dohmae,K.,Nishimune,Y.,Kondo,K.,Kitamura,Y.: "Anemia and mast cell depletion in mutant rat that are homozygous at “white spotting (Ws)" locus." Blood. 78. 1936-1941 (1991)
• [Publications] Katayama,T.,Shiota,K.,Takahashi,M.: "Effects of activin A on anterior pituitary cells fractionated by centrifugal elutriation." Molecular and Cellular Endocrinology. 77. 167-173 (1991)
• [Publications] Ogawa,K.Kurohmaru,M.,Shiota,K.,Takahashi,M.,Nishida,T.Hayashi,Y.: "Histochemical localization of inhibin and activin α, βA and βB subunits in rat gonads." Journal of Veterinary Medical Science. 53. 207-212 (1991)
• [Publications] Saito,S.,Sugino,K.,Yamanouchi,K.,Kogawa,K.,Titani,K.Shiota,K.,Takahashi,M.,Sugino,H.: "Characterization of antisera directed against follistatin/activinbinding protein peptides." Endocrinologia Japonica. 38. 377-382 (1991)
• [Publications] Suzuki,K.,Kamei,T.,Hakamata,Y.,Kikukawa,K.,Shiota,K.,Takahashi,M.: "Basic fibroblst growth factorーlike substance in nuclei of male germ cells undergoing meiosis." Proceeding of the Society for Experimental Biology and Medicine. 198. 728-731 (1991)
• [Publications] Lu,R.ーZ.,Shiota,K.,Tachi,C.,Takahashi,M.: "Histochemical demonstration of activin/inhibin βAー,βBー and αーsubunits in the early embryos and oviducts of different strains of mice." Journal of Reproduction and Development. 38. (1992)
• [Publications] Matsuyama,S.,Shiota,K.,Nashihara,M.,Tachi,C.,Takahashi,M.: "Splenic macrophage enhance luteotropic action of prolactin and luteinizing hormone in rat luteal cell cultures." Endocrinologia Japonica. 39. (1992)
• [Publications] Nishikawa,S.,Kusakabe,M.,Yoshinaga,K.,Ogawa,M.,Hayashi,S.ーI.,Kunisada,T.,Era,T.,Sakakura,T.,Nishikawa,S.ーI.: "In utero manipulation of coat formation by a monoclonal anticーkit antibody:Two distinct waves of cーkitーdependenc during melanocyte development." EMBO Journal. 10. 2111-2118 (1991)
• [Publications] Takenaka,T.,Saga,S.,Miyaishi,O.,Imai,M.,Yokoi,T.,Wajjwalku,W.,Kusakabe,M.,Taguchi,O.,Matsuyama,M.,Hoshino,M.: "Establishment and characterization of immortalized nonーtransplantable mouse mammary cell lines cloned from a MMTVーinduce tumor cell line cultured for a long duration." Nagoya Journal of Medical Science. 53. 9-21 (1991)
• [Publications] Saga,Y.,Tsukamoto,T.,Jing,N.,Kusakabe,M.,Sakakura,T.: "Murine tenascin: cDNA cloning,structure and temporal expression of isoforms." Gene. 104. 177-185 (1991)
• [Publications] 佐藤 英明: "ウシ卵母細胞の成熟と死の制御" 日本畜産学会報. 62. 978-989 (1991)
• [Publications] Sakakibara,R.,Hashida,K.,Tomonaga,N.,Sakai,K.,Ishiguro,M.,Imamura,S.,Ohmatsu,F.,Sato,E.: "A putativ mouse oocyte maturation inhibitory protein from urine of pregnant women:Nーterminal sequence homology with human nonsecrtory ribonuclease." Chem. Pharm. Bull.39. 146-149 (1991)
• [Publications] Sato,E.,Tanaka,T.,Takeya,T.,Miyamoto,H.,Koide,S.S.: "Ovarian glycosaminoglycans potentiate angiogenic activity of epidermal growth factor in mice." Endocrinology. 128. 2402-2406 (1991)
• [Publications] Akabayashi,A.,Kato,T.: "An enzymatic microdetermination method for hydroxylase cofactor." Anal.Lett.24. 1967-1981 (1991)
• [Publications] Akabayashi,A.,Kato,T.: "Microdetermination of 2ーdeoxyglucose and 2ーdeoxyglucose 6ーphosphate to determine glucose utilization rates in single neurons and small CNS regions after injecting nonーtracer amounts of 2ーdeoxyglucose." Journal of Neurochemistry. (1992)
• [Publications] Ookata,K.,Hisanaga,S.,Okano,T.,Tachibana,K.,Kishimoto,T.: "Relocation and distinct subcellular localization of p34^<cdc2>/cyclin B complex at meicsis reinitiation in starfish oocytes." EMBO Journal. 11. (1992)
• [Publications] Lee,G.H.,Nomura,K.,Kanda,H.Kusakabe,M.,Yoshiki,A.,Sakakura,T.,Kitagawa.T.: "Strain specific sensitivity to diethylnitosamineーinduce carcinogenesis is maintained in hepatocytes of C3H/NeNーC57BL/6N chimeric mice." Cancer Research. 51. 3257-3260 (1991)
• [Publications] Yoshiki,A.,Hanazono,M.,Oda,S.ーI.,Wakasugi,N.,Sakakura,T.,Kusakabe,M.: "Developmental analysis of the eye lens Obsolescence (Elo) gene in the mouse:Cell proliferation and Elo gene expression in the aggregation chimera." Development. 113. 1293-1304 (1991)
• [Publications] Tsukamot,T.,Kusakabe,M.,Saga,Y.: "In situ hybridization with nonーradioactive digoxigeninー11ーUTPーlabeled cRNA probes: Loclization of developmentally regulated mouse tenascin mRNAs." International Jouranl of Developmental Biology. 35. 25-32 (1991)
• [Publications] Okamura,N.,Dacheux,F.,Venien,A.,Onoe,S.,Huet,J.ーC.,Dacheux,J.ーL.: "Localization of a maturation dependent epididymal sperm surface antigen recognized by a monoclonal antibody raised against a 135kDa protein in porcine epididymal fluid." Biology of Reproduction. (1992)
• [Publications] Dacheux,J.ーL.,Okamura,N.,Chevrier,C.,Dacheux,F.,Jeulin,C.,Gatti,J.ーL.,Paquignon,M.: "The epididymal environment and sperm maturation." Comparative Spermatology. (1992)
• [Publications] Wada,S.,Okuno,M.,Mohri,H.: "Inner arm dynein ATPase fraction of sea urchin sperm flagella causes active sliding of axonemal outer doublet microtubule." Biochemical and Biophysical Research Communications. 175. 173-178 (1991)
• [Publications] IWamoto,T.,Tsang,A.,Luterman,M.,Dickson,J.,Lamirande,E.,Okuno,M.,Mohri,H.,Gagnon,C.: "Purification and characterization of a sperm motilityーdynein ATPase inhibitor from boar seminal plasma." Molecular Reproduction and Development. 31. 55-62 (1991)
• [Publications] Okuno,M.,Wada,S.,Hayashi,S.,Morisawa,M.: "Enzymatic dissection of the initiation mechanism of flagellar movement in salmonid fish spermatozoa demembranated with Triton Xー100." Cell Motility and Cytoskeleton. (1992)
• [Publications] 北原 聡,大島 博幸: "リプロダクションと夫の年齢" 周産期医学. 21. 1823-1826 (1991)
• [Publications] Abe,S.,Uemura,M.: "How is the flagellar length of mature sperm determined? III. Comparison of initial growth rate of flagella in spermatids from Cynopus and Xenopus in the presence and absence of cycloheximide." Experimental Cell Research. (1992)
• [Publications] Kikuchi,M.,Ishii,S.: "Changes in luteinizing hormone receptors in the granulos and theca layers of the ovarian follicle during follicular maturation in the Japanese quail." General and Comparative Endocrinology. 85. 124-137 (1992)
• [Publications] Ishii,S.,Itoh,M.: "Amplexus induces surge of luteinizing hormone in male toads,Bufo japonicus." General and Comparative Endocrinology. 85. (1992)
• [Publications] Ishii,S.,Wada,M.,Wakabayashi,S.,Sakai,H.,Kurodera,Y.,Yamaguchi,N.,Kikuchi,M.: "Endocrinological studies for artificial breeding of the Japanese ibis,Nipponia nippon,an endangered avian species in Asia." Proceedings of the International Symposium on Environmental and Hormcnal Approaches to Ornithology (1992).(1992)
• [Publications] Okamura,N.,Tajima,Y.,Onoe,S.,Sugita,Y.: "Purification of bicarbonateーsensitive sperm adenylcyclase by 4ーacetamidoー4'ーisothiocyanostilbeneー2,2'ーdisulfonic acidーaffinity chromatoーgraphy." Journal of Biological Chemistry. 266. 17754-17759 (1991)
• [Publications] Okamura,N.,Cnoe,S.,Sugita,Y.,Paquignon,M.,Dacheux,F.,Dacheux,J.ーL.: "Water insoluble fraction of egg yolk maintains porcine sperm motility by activating adenylate cyclase." Molecular Reproduction and Development. 28. 136-142 (1991)
• [Publications] Ishijima,S.A.,Okuno,M.,Mohri,H.: "Zeta potential of human Xー and Yーbearing spermatozoa." International Journal of Andrology. 14. 340-347 (1991)
• [Publications] Ishijima,S.A.,Okuno,M.,Nakagome,Y.,Odagiri,H.,Mohri,T.,Mohri,H.: "Separation of Xー and Yーchromosomeーbearing murine sperm by freeーflow electrophoresis:Evaluation of separation using PCR." Zoological Science. 9. (1992)
• [Publications] Ishijima,S.A.,Okuno,M.,Nakagome,Y.,Nakahori,Y.,Seki,S.,Kaneko,S.,Mohri,H.: "Identification of Xー and Yーchromosomeーbearing human sperm separated by freeーflow electrophoresis using Yーchromosomeーspecific polymerase chain reaction." Biomedical Research. (1992)
• [Publications] Abe,S.,Hiyoshi,H.: "Synthesis of spermーspecific basic nuclear proteins (SPs) in cultured spermatids from Xenopus laevis." Experimental Cell Research. 194. 90-94 (1991)
• [Publications] Hiyoshi,H.,Uno,S.,Yokota,T.,Katagiri,Ch.,Nishida,H.,Takai,M.,Agata,K.,Eguchi,G.,Abe,S.: "Isolation of cDNA for a Xenopus spermーspecific basic nuclear protein (SP4) and evidence for expression of SP4 mRNA in primary spermatocytes." Experimental Cell Research. 194. 95-99 (1991)
• [Publications] Abe,S.,Uno,S.,Asakura,S.,kubokawa,K.: "Inhibition of in vitro spermatogenesis from Xenopus laevis by theoーphylline." Zoological Science. 8. 533-540 (1991)
• [Publications] 岸本 健雄: "新生化学実験講座 第14巻「発生、分化および老化」第13章 卵成熟とcdc2キナ-ゼ" 東京化学同人, (1992)
• [Publications] Ishii,S.: "Gonadotropin in “Vertebrate Endocrinology: Fundamentals and Biomedical Implications",Volume 4,Part B, eds.by P.K.T.Pang, M.P.Schreibman" Academic Press Inc., 33-66 (1991)
• [Publications] 毛利 秀雄: "精子の生物学" 東京大学出版会, 157 (1991)
• [Publications] 毛利 秀雄,森沢 正昭,星 元紀: "精子学" 東京大学出版会, (1992)