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Biochemical and endocrinological analysis of oocytes and embryos

Research Project

Project/Area Number 03670771
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionUniversity of Tokyo

Principal Investigator

TSUTSUMI Osamu  Tokyo University, Dept Ob.Gyn.Assistant Professor, 医学部(病), 講師 (60134574)

Co-Investigator(Kenkyū-buntansha) FUJIWARA Toshihiro  Tokyo University, Dept Ob.Gyn.Assistant, 医学部(病), 助手 (80219063)
AYABE Takuya  Tokyo University, Dept Ob.Gyn.Assistant, 医学部(病), 助手 (50184247)
YANO Tetsu  Tokyo University, Dept Ob.Gyn.Assistant, 医学部(病), 助手 (50182390)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Keywordsoocyte / embryo / glucose / glucose transporter / hexokinase / epidermal growth factor / activin / 卵 / 初期胚 / 卵成熟 / epidermal growth factor(EGF) / グルコ-ス / グルコ-ス取込み能 / カルシウムイオン / 酵素活性
Research Abstract

Microtechniques and highly sensitive methods such as enzymatic cycling, micro-Western analysis, reverse transcription polymerase chain reaction and so on were employed to study these processes. Low hexokinase activity and high activities of enzymes in the phosphate pathway were characteristic of immature oocytes. During maturation, the activities of hexokinase and phosphofructokinase increased significantly. These changes were used to analyze involvement of epidermal growth factor (EGF) and prostaglandins (PG) in oocyte maturation. EGF is shown to stimulate maturation by increasing PG production in granulosa cells. Electro-physiologically, the sensitivity of oocyte to inositol triphosphate increased and Ca^<2+> release system developed during maturation. Progesterone production of oocyte and embryos are shown by enzymatic cycling and other methods using radiometry. This hormone produced by embryos themselves may play a role in embryonic development in intracrine fashion. There is 100-fold increase in glucose uptake from oocyte to blastocyst in mice. A switch in substrate preference of the embryo from pyruvate to glucose during preimplantation development may be explained by increases in the activity of hexokinase and expression of glucose transporter, GLUT1. Hexokinase activities determined by NADP cycling increased 20-fold while expression of GLUT1 assessed by micro-Western method 10-fold. GLUT1 expression was also analyzed by RT-PCR, which indicated that the expression is regulated at transcription level. There is a delay in the developmental changes in glucose uptake, hexokinase activity and GLUT1 expression when the embryos are developed in vitro. It is of interest to note that EGF stimulates these developmental changes in vitro. In conclusion, expanding knowledge in the process of oocyte maturation and embryonic development may serve not only as significant clinical implications for in vitro fertilization techniques but as breakthroughs in gynecology.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (39 results)

All Other

All Publications (39 results)

  • [Publications] O.Tsutsumi,K.Satoh,Y.Taketani,T.Kato: "Determination of enzyme activities of energy metabolism in the maturing rat oocyte." Molecular Reproduction and Development. 33. 333-337 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Morita,O.Tsutsumi,et al.: "Expression and possible function of glucose transporter protein GLUT1 during preimplantation mouse development from oocytes to blastocysts." Biochem.Biophys.Res.Commun.188. 8-15 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Kamei,O.Tsutsumi,et al.: "Intrauterine growth retardation and fetal losses are caused by epidermal growth factor deficiency in mice." American Journal of Physiology. 264. R597-R600 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Sadatsuki,O.Tsutsumi et al.: "Presence and possible function of activin like substance in human follicular fluids." Human Reproduction. 8. 1392-1395 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Sadatsuki,O.Tsutsumi et al.: "Local regulatory effects of activin A and follistatin on meiotic maturation of rat oocytes." Biochem.Biophys.Res.Commun.196. 388-395 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] O.Tsutsumi,Y.Taketani and T.Oka: "The uterine growth promoting action of epidermal growth factor and its function in the fertility of mice." Journal of Endocrinology. 138. 437-443 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 堤 治 他: "生殖医学実験マニュアル" 南江堂, 314 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 堤 治 他: "産婦人科内視鏡手術マニュアル" 金原出版, 115 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] O.Tsutsumi, K.Satoh, Y.Taketani and T.Kato: "Determination of enzyme activities of energy metabolism in the maturing rat oocyte" Molecular Reproduction and Development. 33. 333-337 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Morita, O.Tsutsumi, I.Hosoya, Y.Taketani, Y.Oka and T.Kato: "Expression and possible function of glucose transporter protein GLUT1 during preimplantation mouse development from oocytes to blastocysts" Biochemical and Biophysical Research Communications. 188. 8-15 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] O.Tsutsumi, Y.Taketani and T.Oka: "The uterine growth promoting action of epidermal growth factor and its function in the fertility of mice" Journal of Endocrinology. 138. 437-443 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Kamei, O.Tsutsumi, Y.Kuwabara and Y.Taketani: "Intrauterine growth retardation and fetal losses are caused by epidermal growth factor deficiency in mice" American Journal of Physiology. 264. R597-R600 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Sadatsuki, O.Tsutsumi, R.Yamada, M.Muramatsu and Y.Taketani: "Local regulatory effects of activin A and follistatin on meiotic maturation of rat oocytes" Biochemical and Biophysical Research Communications. 196. 388-395 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Sadatsuki, O.Tsutsumi, R.Sakai, Y.Eto, N.Hayashi and Y.Taketani: "Presence and possible function of activin like substance in human follicular fluids" Human Reproduction. 8. 1392-1395 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] O.Tsutsumi, Y.Morita, I.Hosoya, T.Yano and Y.Taketani: "Delayd expression of hexokinase and glucose transporter causes decreased glucose incorporation in mouse embryos developed in vitro" Serono Symposia Review. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] O.Tsutsumi, T.Yano and Y.Taketani: "Phosphofructokinase activity as a measure of maturation of rat oocytes developed in vivo and in vitro" Hormone Research. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Kamei,O.Tsutsumi,et al.: "Intrauterine growth retardation and fetal losses are caused by epidermal growth factor deficiency in mice." American Journal of Physiology. 264. R597-R600 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Sadatsuki,O.Tsutsumi et al.: "Presence and possible function of activin like substance in human follicular fluids." Human Reproduction. 8. 1392-1395 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Sadatsuki,O.Tsutsumi et al.: "Local regulatory effects of activin A and follistatin on meiotic maturation of rat oocytes." Biochem.Biophys.Res.Commun.196. 388-395 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] O.Tsutsumi,Y.Taketani and T.Oka: "The uterine growth promoting action of epidermal growth factor and its function in the fertility of mice." Journal of Endocrinology. 138. 437-443 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] O.Tsutsumi,Y.Taketani and T.Oka: "Evidence for the involvement of epidermal growth factor in fertility decline in aging female mice." Hormone Research. 39(suppl 1). 32-36 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] O.Tsutsumi,et al.: "Delayed expresson of hexokinase and glucose transporter causes decreased glucose incorporation in mouse embryos developed in vitro." Serono Symposia Review. (199)

    • Related Report
      1993 Annual Research Report
  • [Publications] 堤 治他: "生殖医学実験マニュアル" 南江堂, 314 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 堤 治他: "産婦人科内視鏡手術マニュアル" 金原出版, 115 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] O.Tsutsumi,K.Satoh,Y.Taketani,T.Kato: "Determination of enzyme activities of energy metabolism in the maturing rat oocyte." Mol.Reprod.Dev.33. 333-337 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Morita,O.Tsutsumi,et al.: "Expression and possible function of glucose transporter protein GLUT1 during preimplantation mouse development from oocytes to blastocysts." Biochem.Biophys.Res.Comm.188. 8-15 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Kamei,O.Tsutsumi,Y.Kuwabara,Y.Taketani: "Intrauterine growth retardation and fetal losses are caused by epidermal growth factor deficiency in mice." Am.J.Physiol.(1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] O.Tsutsumi,Y.Taketani,T.Oka:"Evidence for the involvement of epidermal growth factor in fertility decline in aging female mice." Hormone Research. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 細谷 岩生,堤 治,他: "マウス初期胚糖取込み能への成長因子の影響" 哺乳動物卵子学会雑誌. 9. (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 森田 豊,堤 治,武谷 雄二,水野 正彦: "卵の成熟分化にともなうエネルギー代謝の発達" 産科と婦人科. 59. 871-876 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.Mizuno,S.G.Liang,S.Ishihara,O.Tsutsumi,Y.Taketani: "Recent advances in ovarian function:Basic and clinical researches." Fujimoto & Mizuno(eds) Christengraf,Rome, 12 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 堤 治: "生殖医学実験マニュアル" 南江堂, 16 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] TーM Kuo,Y. Taketani,T.Ayabe,O.Tsutumi M. Mizuno: "Stimulatory effect of epidermal growth factor on the development of mouse early embryo in vitro." Endoctinol. Japon.38. 485-490 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] O. Tsutsumi,K. Satoh,M. Mizuno,T. Kato: "Determination of enzyme activities of energy metabolism in maturing rat oocyte" J. Reprod. Fert.

    • Related Report
      1991 Annual Research Report
  • [Publications] 堤 治,矢野 哲,綾部 琢哉,藤原 敏博,細谷 岩生: "卵成熟機構の生化学的解析" 日本受精着床学会雑誌. 8. 14-17 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 成 鑑非,堤 治,藤原 敏博,三橋 直樹,水野 正彦: "体外受精時卵胞液に検出されるhCG濃度と卵成熟の関連" 日本産科婦人科学会雑誌. 43. 789-790 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 細谷 岩生,堤 治,藤原 敏博,三橋 直樹,水野 正彦: "微小培養法を用いた単一卵分析によるマウス初期胚糖取込み能発達の解析" 日本産科婦人科学会雑誌. 43. 1579-1580 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 百枝 幹雄,堤 治,水野 正彦也: "高感度尿中LH測定キットの臨床有用性の検討" 日本受精着床学会雑誌. 8. 67-71 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 我妻 尭,前原 澄子 編,堤 治: "助産学講座2生殖の形態・機能「卵巣機能」" 医学書院, (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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