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Effect of Nitric Oxide on the Control of Ovarian functions

Research Project

Project/Area Number 06671633
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionTokyo Medical and Dental University

Principal Investigator

KUBOTA Toshiro  Tokyo Medical and Dental Univ.Lecturer, 医学部, 講師 (50126223)

Co-Investigator(Kenkyū-buntansha) ASO Takeshi  Tokyo Medical and Dental Univ.Professor, 医学部, 教授 (60093176)
KAMADA Shusaku  Tokyo Medical and Dental Univ.Lecturer, 医学部, 講師 (80169606)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1994: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsNitric Oxide / Porcine granulosa cell / Steroidogenesis / LNMMA / NO synthase / Cell culture / Aromatase activity / cGMP / NO合成酵素 / cGMP / ゴナドトロピン / 細胞増殖
Research Abstract

Nitric oxide (NO) has been recently implicated in a variety of physiological precesses, including reproductive functions. The present study was undertaken to investigate whether NO might significantly induce any change in steroidogenesis in cultured porcine granulosa cells (PGC).
Cells were obtained from medium follicles by the needle aspiration method. After pre-incubation, cells were incubated with N^G-monomethyl-L-arginine (LNMMA), a competitive blocker of NO synthase (NOS), and the NO donor, NOC 18, either alone or in the presence of FSH (200ng/ml) / hCG (5IU/ml). Estradiol (E_2) and progesterone (P) concentrations in media were measured by EIA.LNMMA (0.01- 1 mM) significantly (P<0.05) stimulated the basal E_2 release and dosedependently enhanced the E_2 and P release from FSH-stimulated PGC for 24-h culture. LNMMA also significantly (P<0.001) suppressed the NO^- concentrations in media, measured by chemiluminescence. NOC 10 (0.1 - 1 mM) significantly (P<0.01) suppressed the E_2 release from basal and gonadotropin stimulated PGC for 2-h culture. Cells were immunostained with an avidin-biotin-peroxidase (ABC) kit, and the localization of constitutive (c) NOS and inducible (i) NOS was immunohistochemically comfirmed. Additionnally, we evaluated the possible involvement of cGMP and aromatase activity on NO-induced inhibition of steroidogenesis. The cGMP analog, 8-bromo-cGMP,had no significant effect on the accumulation of E_2 and P in basal and gonadotropin simulated PGC during 24-h culture. However, NOC 18 (0.1 - 1 mM) significantly (P<0.01) inhibited the aromatase activity of basal and gonadotropin stimulated PGC,measured by a modified tritiated water method.
These results show that no inhibits E_2 secretion independent of cGMP by directly inhibiting aromatase activity in moderately mature PGC,and suggest that NO is an important local modulator of the follicular development.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Kubota T. et al.: "Autocrine/Paracrine function of transforming growth factor-Bl in porcine granulosa cells" Human Reproduction. 9. 2118-2122 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M., Kubota T. et al.: "The effect of nitric oxide on steroidogenesis in culture porcine granulosa cells" J. Assisted Reproduction and Gynetics. 12. 13- (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kubota T. et al.: "Endothelin synthesis and receptors in human endo-metrium throughout the rormal menstroual cycle." Human Reprduction. 10. 2204-2208 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kamada S. et al.: "The role of endoehelin-regulating human granulosa cell proliferation and steroidogonesis in vitro" Journal of Clinical Endocrinology and Metabolism. 80. 3708-3714 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 増田美香子、久保田俊郎他: "ブタ顆粒膜細胞のステロイド産生能及び細胞増殖能に及ぼす一酸化窒素の影響" 日本産科婦人科学会雑誌. 47. 277- (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 増田美香子、久保田俊郎他: "Sodium Nitroprussicdeの培養ブタ顆粒膜脂肪ステロイド産生能に及ぼす影響について" 日本内分泌学会雑誌. 71. 366- (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M. Kubota T. et al: "New Achievements in Research of Ovarian Function Froneiers in Endocrinology, volume 13" Ares-Serono Sympisia Publication, Rome Iealy, 347 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M., Kubota T. et al.: "IX World Congress on IN VITRO FERTILIZATION and ASSiSTED REPrOPUCTION" Mondu zzi Editore, Bologna Italy, 772 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kubota, T., Kamada, S., Taguchi, M.and Aso, T: "Autocrine/paracrine function of transforming growth factor-beta1 in porcine granulosa cells." Human Reproduction. 9 : (11). 2118-2122 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kubota T., Taguchi M., Kamada S., Imai T., Hirata Y.and Aso T: "Endothelin synthesis and receptors in human endometrium throughout the normal menstrual cycle." Molecular Human Reproduction. 1, (Human Reproduction 10). 2204-2208 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kamada S., Blackmore P., Kubota T., Oehninger S., Asada Y., Gordon K.and Hodgen G.D.: "The role of endothelin-1 regulating human granulosa cell proliferation and steroidogenesis in vitro." Journal of Clinical Endocrinology and Metabolism. 80. 3708-3714 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M., Kubota T., Kamada S.and Aso T: "The effect of nitric oxide on steroidogenesis in culture porcine granulosa cells." Journal of Assisted Reproduction and Gynetics. 12 (3). 13 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M., Kubota T., Kamada S., and Aso T: "Nitric oxide inhibits steroidogenesis in porcine granulosa cells." 10th International Congress of Endocrinology, June 12-15, San Francisco, CA,USA,1996. (submitting.).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M., Kubota T., Kamada S., and Aso T: The effect of nitric oxide on steroidogenesis in cultured porcine granulosa cells. New Achievements in Research of Ovarian Function. Frontiers in Endocrinology, Volume 13.Fujimoto S., Straus III J.F., Hsueh A.J.W.and Tanaka T., Serono Symposia Publication, Rome Italy, 280-284 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Masuda M., Kubota T., Kamada S.and Aso T: The effect of nitric oxide on steroidogenesis in cultured porcine granulosa cells. In IX World Congress on IN VITRO FERTILIZATION AND ASSISTED REPRODUCTION.Aburumieh H., Bernat E Fischl F.et al.editors, Monduzzi Editore, Bologna Italy, 159-163 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Kubota T.,et al.: "Autocrine/paracrine function of transforming growth factor-B1 in purcine granulosa cells" Human Reproduction. 9. 2118-2122 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kubota T.,et al.: "Endothelin synthesis and receptors in human endometrium throughout the normal menstrual cycle" Human Reproduction. 10. 2204-2208 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Komada S.,et al.: "The role of endothelin-1 regulating human granulosa cell proliferation and steroidgenesis in vitro" Journal of Clinical Endocrinology and Metabolism. 80. 3708-3714 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Masuda M.,Kubota T.,et al.: "The effect of nitric oxide on steroidogenesis in culture porcine granulosa cells." J.Assisted Reproduction and Gynetics.12. 13 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 増田 美香子,久保田 俊郎,他: "ブタ顆粒膜細胞のステロイド産生能及び細胞増殖能に及ぼす一酸化窒素の影響" 日本産科婦人科学会雑誌. 47. 277 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 増田 美香子,久保田 俊郎,他: "Sodium nitroprussideの培養ブタ顆粒膜細胞ステロイド産生能に及ぼす影響について" 日本内分泌学会雑誌. 71. 366 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Masuda M,Kubota T,et al.: "New Achievements in Research of Ovarian Function Frontiers in Endocrinology,volume13" Ares-Serono Symposia Publication,Rome Italy, 347 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Masuda M,Kubota T,et al.: "IX World Congress on IN VITRO FERTILIZATION and ASSISTED REPRODUCTION" Monduzzi Editore,Bologna Italy, 772 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Kubota et al.: "Autocrine/paracrine function of transfurming growth factor-β1 in porcine granulosa cells" Human Reproduction. 9. 2118-2122 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Kubota et al.: "Endothelin-1 as a local ovarian regulator in porcine granulosa cells" Hormone Research. 41. 29-35 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Masuda,T.Kubota et al.: "The effect of nitric oxide on steroidogenesis in porcine granulosa cells." 3rd International Symposium on Warian Function. (Abseract). 38 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 増田美香子,久保田俊郎 他: "ブタ顆粒膜細胞のステロイド産生能および細胞増殖能におよぼすNO影響" 日本不妊学会雑誌. 39. 530 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 増田美香子,久保田俊郎 他: "ブタ顆粒膜細胞のステロイド産生能および細胞増殖能に及ぼす一酸化窒素(NO)の影響" 第47回日本産科婦人科学会総会. 4月(発表予定). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Kubota et al: "Frontiers in Endocrinology vol.4 Pergpectives on assisted reproduction" Ares-Serono Symposia Publication,Rome,I, 749 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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