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1999 Fiscal Year Final Research Report Summary

Studies on production of pig embryos from growing oocytes

Research Project

Project/Area Number 09556061
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Applied animal science
Research InstitutionKOBE UNIVERSITY

Principal Investigator

KATO Seishiro  Kobe University, Faculty of Agriculture, Professor, 農学部, 教授 (90026386)

Co-Investigator(Kenkyū-buntansha) MIYANO Takashi  Kobe University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (80200195)
MIYAKE Masashi  Kobe University, Graduate School of Science and Technology, Professor, 大学院・自然科学研究科, 教授 (60093316)
KANNAN Yasuyuki  Kobe University, Faculty of Agriculture, Professor, 農学部, 教授 (60031192)
TOMINAGA Keiichiro  Hyogo Prefectural Agricultural Institute, Senior Investigator, 主任研究員
HARAYAMA Hiroshi  Kobe University, Graduate School of Science and Technology, Associate Professor, 大学院・自然科学研究科, 助教授 (30281140)
Project Period (FY) 1997 – 1999
Keywordspig / oogenesis / folliculogenesis / oocyte growth / oocyte maturation / in vitro culture / in vitro fertilization / embryo production
Research Abstract

Several hundred thausand primordial follicles are contained in the ovaries of domestic animals. The development of in vitro culture systems for the growth of small oocytes in primordial or small follicles is expected to provide a new source of a large population of oocytes for livestock production. This study was conducted to develop Culture systems for small oocytes from pig ovaries.
Preantral follicles containing oocytes with various sizes were isolated from pig ovaries and cultured in collagen gel for up to 16 days in the presence of serum, FSH and oestradiol. Waymouth MB7 52/1 was used as a basic culture medium for oocyte growth. After culture, the oocytes enclosed by one or two layers of cumulus cells were cultured for meiotic maturation and inseminated with spermatozoa. The results obtained were as follows.
1. Pig growing oocytes grew to their final size, acquired meiotic competence du ring in vitro culture, and were penetrated by spcermatozoa in vitro, although there was a very lo … More w level of success in fertilization. In contrast to the growing oocytes, no culture methods were available for much smaller oocytes. We tried to culture primordial follicles which contained small oocytes 30 μm in diameter, but the integrity of the follicles was soon broken and the follicles survived for only a few days.
2. Using culture systems, we demonstrated that granulosa cells regulate oocyte growth, and that antrum formation of granulosa cells was induced only by meiotic-competent oocytes, indicating that the oocytes secrete a substance(s) that induces differentiation of granulosa cells.
3. Of four glycosaminoglycans examined, hyalronic acids and chondroitin sulfate A supported the development of in vitro-matured and -fertilized pig oocytes to the blastocyst stage.
4. In addition, degradation of cyclin B1 molecules and MAP kinase deph osphorylation during fertilization, localisation of phosphorylated MAP kinase during the transition from meiosis I to meiosis II, assembly and transformation of the spindle during the progression through meiotic cell cycle, and the level and localization of CENP-E and kinetochore numaber during maturation were examined. Less

  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Miyano,T.: "Acquisition of meiotic competence during oocyte growth in mammals"J.Mamm.Ora Res.. 14. 117-124 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shen,X: "In vitro antrum formation of oocyte-cumulus-granulosa cell complexes from pig early antral follicles"J.Mamm.Ora.Res.. 14. 183-190 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shen,X: "Promotion of follicular antrum formation by pig oocytes in vitro"Zyote. 6. 47-54 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kano,K.: "Effects of glycosaminoglycans on the development of in vitro-matured and-fertilized porcine oocytes to the blastocyst stage in vitro"Biol.Reprod.. 58. 1226-1232 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Harayama,H.: "Effects of calcium and bicarbonate on head-to-head agglutination in ejaculated boar spermatozoa"Reprod.Fertil.Dev.. 10. 445-450 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyano,T.: "In vitro culture of pig oocytes collected from early antral follicles"J.Mamm.Ora Res.. 15. 161-166 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lee,J.: "Thyrosine phosphorylation of p34^<cdc2> in metaphase II-arrested pig oocytes results in pronucleus formation without chromosome segregation"Mol.Reprod.Dev.. 52. 107-116 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Harayama,H: "Changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during capacitation in vitro"Reprod.Fertil.Dev.. 11. 193-199 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lee,J.: "Localisation of phosphorylated MAP kinase during the transition from meiosis I to meiosis II in pig oocytes"Zygote. 8. 119-125 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyano,T.: "Degradation of pig cyclin B1 molecules precedes MAP kinase dephosphorylation during fertilization of the oocytes"Zygote. 8. 153-158 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lee,J.: "Spindle formation and dynamics of r-tubulin and nuclear mitotic apparatus protein distribution during meiosis inpig and mouse oocytes"Biol.Reprod.. 62. 1184-1192 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Harayama,H.: "Biochemical characterization of sialoprotein "Anti-Agglutinin" purified from boar epididymal and seminal plasma"Mol.Reprod.Dev.. 55. 96-103 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kure-bayasi,S.: "Successful implantation of in vitro-activated oocytes in the pig"Theriogenology. 53. 1105-1119 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lee,J.: "Specific regulation of CENP-E and kinetochores during meiosis I/meiosis II transition in pig oocytes"Mol.Reprod.Dev.. 56. 51-62 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Jsuji-Goto,Y.: "Meiotic resumption of pig oocytes by cAMP-dependent protein kinase inhibitor H89"J.Mamm.Ora Res.. 17. 58-63 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miyano, T.: "Acquisition of meiotic competence during oocyte growth in mammals"J.Mamm.Ova Res.. 14. 117-124 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shen, X.: "In vitro antrum formation of oocyte-cumulus-granulosa cell complexes from pig early antral follicles"J.Mamm.Ova Res.. 14. 183-190 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shen, X.: "Promotion of follicular antrum formation by pig oocytyes in vitro"Zygote. 6. 47-54 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kano, K: "Effects of glycosaminoglycans on the development of in vitro-matured and -fertilized porcine oocytes to the blastocyst stage in vitro"Biol. Reprod.. 58. 1226-1232 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Harayama, H.: "Effects of calcium and bicarbonate on head-to-head agglutination in ejaculated boar spermatozoa"Reprod. Fertil. Dev.. 10. 445-450 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miyano, T.: "In vitro culture of pig oocytes collected from early antral follicles"J.Mamm. Ova Res.. 15. 161-166 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lee, J.: "Thyrosine phosphorylation of p34^<cdc2> in metaphase II-arrested pig oocytes results in pronucleus formation without chromosome segregation"Mol. Reprod. Dev.. 52. 107-116 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Harayama, H.: "Changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during capacitation in vitro"Reprod. Fertil. Dev.. 11. 193-199 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lee, J.: "Localisation of phosphorylated MAP kinase during the transition from meiosis I to meiosis II in pig oocytes"Zygote. 8. 119-125 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miyano, T.: "Degradation of pig cyclin B1 molecules precedes MAP kinase dephosphorylation during fertilization of the oocytes"Zygote. 8. 153-158 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lee, J.: "Spindle formation and dynamics of r-tubulin and nuclear mitotic apparatus protein distribution during meiosis inpig and mouse oocytes"Biol. Reprod.. 62. 1184-1192 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Harayama, H.: "Biochemical characterization of sialoprotein "Anti-Agglutinin" purified from boar epididymal and seminal plasma"Mol. Reprod. Dev.. 55. 96-103 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kure-bayashi, S.: "Successful implantation of in vitro-activated oocytes in the pig"Theriogenology. 53. 1105-1119 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lee, J.: "Specific regulation of CENP-E and kinetochores during meiosis I / meiosis II transition in pig oocytes"Mol. Reprod. Dev.. 56. 51-62 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsuji-Goto, Y.: "Meiotic resumption of pig oocytes by cAMP-dependentprotein kinase inhibitor H89"J.Mamm. Ova Research.. 17. 58-63 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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