1997 Fiscal Year Final Research Report Summary
active substance regulating differentiation of germ lines
| Project/Area Number |
08456147
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Tohoku University (1997)
The University of Tokyo (1996) |
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Principal Investigator |
SATO Eimei Tohoku Univ.Fac.Agri., Professor, 農学部, 教授 (80093243)
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| Project Period (FY) |
1996 – 1997
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| Keywords | fertilization / oogenesis / spermatogenesis / oocytes / sperm / primordial germ cell / embryos / in vitro culture |
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| Research Abstract |
1) RT/PCR technique coupled with Southern blot hybridization analysis showed that highest level of FasL mRNA was demonstrated in murine ovaries and granulosa cells 1 day after the administration of PMSG,while the level of FasL mRNA became very weak on the day 5, respectively. The observed gradual decrease in FasL mRNA could not be attributed to a generalized degradation of cellular RNA during atresia, as evidenced by the presence of constitutive expression of elongation factorla (EF-1a) mRNA in murine ovaries and granulosa cells treated with PMSG.Furthermore, in situ hybridization analysis with a FasL-specific probe confirmed that FasL was specifically localized in the granulosa cells of most follicles and its expression was regulated by PMSG administration.
2) We found the the inactive MAPK was already present in immature oocytes arrested at the GZ stage and that this inactive kinase was localized exclusively in the cytoplasm. At the GZ/M transition stage, part of the MAPK moved into the germinal vesicle (GV) before germinal vesicle breakdown (GVBD). In addition, immunoblot analysis showed that the nuclear MAPK existed in an active form. To determine whether this active could induce GVBD,we microinjected active MAPK into immature porcine oocytes. The active MAPK injected into the cytoplasm was quickly inactivated and could not accelerate GVBD.In contrast, MAPK injection into the GV markedly accelerated GVBD
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[Publications] Matsumoto, H., Shoji, N., Sugawara, S., Umezu, M., Sato, E.: "Microscopic analysis of enzyme activity, mitochondrial distribution and hydrogen peroxide in two-cell rat embryos." J.Reprod.Fert.(in press).
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