| Project/Area Number |
05660334
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied animal science
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| Research Institution | KINKI UNIVERSTIY |
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Principal Investigator |
IRITANI Akira KINKI UNIVERSITY,FACULTY OF BIOLOGY ORIENTED SCIENCE & TECHNOLOGY,PROFESSOR, 生物理工学部, 教授 (80026385)
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| Co-Investigator(Kenkyū-buntansha) |
OISHI Takeshi KINKI UNIVERSITY,FACULTY OF AGRICULTURE,ASSOCICTED PROFFSSOR, 農学部, 助教授 (00088189)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Microfertilization / Inmotile sperm / Embryo development / 外来遺伝子導入技術 |
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| Research Abstract |
The technique of microfertilization is a promising tools for investigation of the events involved in normal sperm-egg interactions and establishement of fertilization with immotlile spermatozoa. In case of electroporation or lipofectin treatment for exogenous DNA transplantation, movement of spermatozoa was damaged severly. In our experiments, the possibility of fertilization of pig or rooster oocytes matured in culture by microfertilization of single spermatozoon into cytoplasm was examined.
Ejaculated spermotozoa washed once and killed by freeze-thawing were used as a dead sperm. Spermatozoa incubated for 10 hours in m-Tyrode solution were used as a weak motile sperm. Fresh ejaculated spermatozoa were used as a good motile sperm. Sixty four percent of the injected oocytes survived, and 52% of them were fertilized. There was no difference of fertilization rate among the treatments for spermatozoa. Injection volume of sperm and seminal plasm affected the viability of injected oocytes. Seminal plasm itself was known as a activation factor for mature oocyte. However even small amount of that had lethal effect for the injectedoocyte. It may be completely different from an oocyte activating factor extracted from spermatozoa.
Poultry spermatozoa has an ability of pronuclear formation in hamster oocytes. It is clear that there was a possibility for normal fertilization by sperm injection.
According to our results, this sperm injection method is useful for embryo production for spermatozoa with poor motility and it will be applicable for sperm transmittted DNA transfer.
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