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Isolation and Chaacterization of Human Inhibin in Follicular Fluid and Production of recombinant Inhibin.

Research Project

Project/Area Number 04670987
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionKitasao Univ.Sch.of Vet.Med.and Anim.Sci (1993)
Gunma University (1992)

Principal Investigator

HASEGAWA Yoshihisa  Kitasato Univ.Dept.Lab.Ani.Sci.Pro, 獣医畜産学部, 教授 (40092001)

Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsHuman Inhibin / CHO cell / Isoation / IRMA / Activin / ヒト卵胞液 / インヒビンの精製 / リコンビナントインヒビン / 免疫吸着クロマト
Research Abstract

Immunoaffinity chromatography is by far the most useful and successfully applied method for purification of the inhibin in follicular fluid. Therefore, we tried to develop the immunoaffinity chromatography with polyclonal antibody to 32-KDa bovine inhibin purified by the improved methods on large scale. Human inhibins were purified from 10 liter human follicular fluid by various kinds of chromatography including Matrex gel red A, immunoaffinity chromatography, gel chromatography and revesed-phase HPLC.
On SDS-PAGE under nonreducing conditions, bioactive inhibin protein had two band migrating at MW 34,000 and 29,000 and similarly, immunoreactive inhibin protein had two bands migrating at MW 30,000 and 26,000, respectively.
cDNA clone encoding human inhibin alpha-and betaB subunits were isolated from a human testicular cDNA library. The cDNAs were linked to an expression vector and transfected into Chinease hamsteer cells (CHO cells). The cells transfected with the betaB subunit cDNA alone and with both the alpha and betaB subunit cDNAs secreted betaB dimer and inhibin B and betaB homodimer could evoke their biological actions on the FSH secretion from rat pituitary cultured cells.

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (36 results)

All Other

All Publications (36 results)

  • [Publications] H.Kaneko,M.Yoshida,Hara,K.Taya,K.Arai,S.Sasamoto,et al: "Involvement of inhibin in the regulation of FSH secretion in prepubertal bulls." Journal of Endocrinology. 137. 15-19 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Nakamura,T.Minegishi,Y.Hasegawa,K.Nakamura,et al: "Effect of an activin A on follicle-stimulating hormone(FSH) receptor messenger ribonucleic acid levels and FSH receptor---" Endocrinology. 133. 538-544 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Yasuda,K.Inoue,H.Shibata,T.Takeuchi,Y.Eto,Y.Hasegawa: "Existence of activin-A in A- and D-cells of rat pancreatic islet." Endocrinology. 133. 624-630 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Yasuda,T.Mine,H.Shibata,Y.Eto,Y.Hasegawa,et al: "Activin A:An autocrine inhibitor of initiation of DNA synthesis in rat hepatocytes." J.Clin.Invest.92. 1491-1496 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Matsumoto,Y.Ishibashi,T.Ohtaki,Y.Hasegawa,et al: "Newly established murine pituitary folliculo-stelite-like cell line(TtT/GF) secretes potent potent pituitary glandlar cell---" Biochem.Biophys.Res.Commun.194. 909-915 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 長谷川喜久: "インヒビン・アクチビン" 産婦人科の実際. 42. 1835-1839 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Hasegawa,K.Miyamoto,H.Sugino,K.Takio,M.Inoue,Y.Ibuki: "Progress with human and rat inhibin characterization.in:Frontiers in Endocrinology" Ares Serono Symposia, 313(5-24) (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 長谷川喜久: "繁殖のホルモン,加藤征史郎編「家畜繁殖学」" 朝倉書店(印刷中), (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Doi, M.Igarashi, Y.Hasegawa, Y.Eto, H.Shibai, T.Miura, and Y.Ibuki: "In vivo action of activin-A on puitart-gonadal system. Endocrinology" 130. 139-144 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Sugino, T.Nakamura, K.Takio, K.Miamoto, Y.Hasegawa, M.Igarashi, K.Titani and H.Sugino: "Purification and charact6erization of high molecular weight forms of inhibin from bovine follicular fluid. Endocrinology" 130. 789-796 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Andoh, H.Mizunuma, Y.Hasegawa, K.Yamada, M.Michshita and Y.Ibuki: "Immunoreactive inhibin levels as a marker of luteal function" Hormone Research. 37(suppl 1). 37-40 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Takeuchi, M.Seki, Y.Hasegawa, K.Tuchiya, M.Itoh, I.Itoh, H.sHINOZAKI, T.Jujo, Y.Ibuki and M.Igarashi: Hormone Research. 37(suppl 1). 69-74 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Nakamura, Y.Hasegawa, K.Sugino, K.Kogawa, K.Titani and H.Sugino: "Follistatin inhibits activin-induced differentiation of rat follicular granulosa cells in vitro" Biochemica et Biophysica Acta. Vol.1135. 103-109 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Amano, K.Okuzawa, T.Yanagisawa, Y.Hasegawa and K.Aida: "Immunoreactive gonadotropin-releasing hormone (Ir-GnRh) in the carp Brain" Nippon Suisan Gakkaishi. 58. 593 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Kaneko, Y.Nakanishi, G.Watanabe, K.Taya, S.Sasamoto and Y.Hasegawa: "Evidence that inhibin is an important factor in the regulation of FSH secretion during the mid-luteal phase in the cows" Journal of Endocrinology. 136. 35-41 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Kaneko, M.Yoshida, Y.Hara, K.Taya, K.Araki, G.Watanabe, S.Sasamoto and Y.Hasegawa: "Involvement of inhibin in the regulation of FSH secretion in prepubertal bulls." Journal of Endocrinology. 137. 15-19 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Shinozaki, I.Ito, Y.Hasegawa, K.Nakamura, S.Igarashi, M.Nakamura, K.Miyamoto, Y.Etoh Y.Ibuki and T.Minegishi: "Cloning and sequencing of a rat type II activin receptor. FEBS Letter" 132. 53-56 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Nakamura, T.Minegishi, Y.Hasegawa, K.Nakamura, S.Igarashi, I.Itoh, H.H.shinozaki, K.Miyamoto, Y.Eto and Y.Ibuki: "Effectof an activin A on FSH receptor messenger ribonucleic acid levels and FSH receptor expression in cultured rat granulosa cells" Endocrinology. 133. 538-544 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Yasuda, K.Inoue, H.Shibata, T.Takeuchi, Y.Eto, Y.Hasegawa, N.Sekine, Y.Totsuka, T.Mine, E.Ogata and I.Kojima: "Existence of activin A in A- and D-cells of rat pancreatic islet" Endorinology. 133. 624-630 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Amano, K.Aida, N.Okumoto and Y.Hasegawa: "Changes in levels of GnRH in the brain and pituitary and GTH in the pituitary in male masu salmon, Oncorhynchus masou, from hatching to maturation" Fish Physiol. and Biochem.11. 233-240 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Yasuda, T.Mine, H.Shibata, Y.Eto, Y.Hasegawa, T.Takeuchi, S.Asano and I.Kojima: "Activin A : An autocrine inhibitor of initiation of DNA synthesis in rat hepatocytes" Journal Clinical Investigation. 92. 1491-1496 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Matsumoto, Y.Ishibashi, T.Ohtaki, Y.Hasegawa, C.Koyama and K.Inoue: "Newly established murine pituitary folliculostellate-like cell line(TtT/GF) secretes potent pituitary glandlar cell survival factors, one of which corresponds to metalloproteinase inhibitor" Biochemical Biophysical Research Communications. 194. 909-915 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] H.Kaneko,M.Yoshida,Y.Hara,K.Taya,K.Arai,: "Involvement of inhibin in the requlation of FSH secretion in prepubertal bulls." Journal of Endocrinology. 137. 15-19 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Nakamura,T.Minegishi,Y.Hasegawa,K.Nakamura,: "Effect of an activin A on follicle-stimulating hormone(FSH)receptor messenger ribonucleic acid levels and FSH receptor---" Endocrinology. 133. 538-544 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] H.Yasuda,K.Inoue,H.Shibata,T.Takeuti,Y.Eto,: "Existence of activin-A in A- and D-cells of rat pancreatic Islet." Endocrinology. 133. 624-630 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] H.Yasuda,T.Mine,H.Shibata,y.Eto,Y.Hasegawa: "Activin A: An autocrine inhibitor of initiation of DNA synthesis in rat hepatocytes." J.Clin.Invest.92. 1491-1496 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] H.Matsumoto,Y.Ishibashi,T.Ohtaki,Y.Hasegawa,: "Newly established murine pituitary Folliculo-stelite-like cell line(TtT/GF)secretes potent pituitary glandlar cell-----" Biochem.Biophys.Res.Commun.194. 909-915 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 長谷川喜久: "インヒビン・アクチビン" 産婦人科の実際. 42. 1835-1839 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.hasegawa,K.Miyamoto,H.sugino,k.Takio,M.Inoue,: "Progress with human and rat inhibin characterization.in;Frontiers in Endocrinology" Ares Serono Symposia, 313(5-24) (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 長谷川喜久: "繁殖のホルモン,加藤征史郎編「家畜繁殖学」" 朝倉書店, (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Sugini,T.Nakamura,K.Takio,K.Miyamoto,Y.Hasegawa,M.Igarashi: "Purification and characterization of high molecular weight forms of inhibin from bovine follicular fluid." Endocrinology. 130. 789-796 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Nakamura,Y.Hasegawa,K.Sugino,K.Kogawa,K.Titani,H.Sugino: "Follistatin inhibits activin-induced differentiation of rat follicular granulosa cells in vitro." Biochimica et Biophysica Acta. 1135. 103-109 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Andoh,H.Mizunuma Y.Hasegawa,K.Yamada M.Michishita,Y.Ibuki: "Association of immunoreactive-inhibin levels with luteal function." Hormone Research. 37. 37-40 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Takeuchi,M.Seki Y.Hasegawa,K.Tsuchiya M.Itoh,I.Itoh,H.Shinozaki: "Further observations on serial serum immunoreactive inhibin levels in the luteal phase and early gestation after ovarian stimulation for in vitro fertilization and embrio transfer." Hormone Research. 37. 69-74 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Shinozaki,I.Itoh,Y.Hasegawa,K.Nakamura,S.Igarashi,M.Nakamura: "Cloning and Sequencing of a rat type II activin receptor." FEBS letters. 312. 53-56 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Kaneko,Y.Nakanishi,K.Taya,H.Kishi,G.Watanabe S.Sasamoto,Y.Hasegawa: "Evidence that inhibin is an important factor in the regulation of FSH secretion during the mid-luteal phase in cows." Journal of Endocrinology. 136. 35-41 (1993)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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