2003 Fiscal Year Final Research Report Summary
Basic study for the production of nock-out pig by nucleiar transfer using recombinant somatic cell
| Project/Area Number |
12480248
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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 |
Laboratory animal science
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
MORI Tadashi Hokkaido Univ., Grad.School of Agriculture, Inst., 大学院・農学研究科, 助手 (30230072)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Yoshiyuki Hokkaido Univ., Grad.School of Veterinary Med., Prof., 大学院・獣医学研究科, 教授 (70167485)
MATSUMOTO Kenichi Hokkaido Univ., Grad.School of Pharm.Sci., Asso.Prof., 大学院・薬学研究科, 助教授 (30202328)
YOSHIDA Ikuya Hokkaido Univ., Center of Advan Sci.and Tech., Inst., 先端科学技術研究センター, 助手 (90240275)
SUZUKI Keita Hokkaido Univ., Field Scientific Center, Inst., 北方生物圏フィールド科学センター, 助手 (60261335)
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| Project Period (FY) |
2000 – 2003
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| Keywords | Pig / Oocytes / Embryos / IP3 / Ca^<2+> concentration / PhospholipaseCζ / Nuclear transfer / EGFP gene |
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| Research Abstract |
Inositol 1,4,5-trisphosphate (IP3) is considered to be important for activation of mammalian oocytes at the time of fertilization, and that the activation induces a rise in intracellular Ca^<2+> concentration. Therefore, IP3 can be useful as an artificial activator of oocytes. In this study, we injected IP3 into porcine matured oocytes and demonstrated that injected IP3 promotes the development of the matured oocytes and that their developmental ability is positively correlated with the rise in Ca^<2+> concentration induced by IP3. Naturally, mammalian oocytes are activated with sperm penetration, and it is suggested that some unknown sperm factor must activate the oocytes. For several years, it has been argued what is a real sperm factor to activate oocytes. Recently, phospholipaseC(PLC)ζ cDNA is cloned and is expected to be the most useful candidate to activate the oocytes. We originally succeed to clone the porcine PLCζcDNA. In addition to these activation works, and finally in vitro developmental study of porcine nuclear transfer embryos from foreign gene-transfected somatic cells was investigated. We used enhanced green fluorescent protein(EGFP) gene as a foreign gene maker. The result indicated that EGFP gene transfected into oocytes has no deleterious effect in the embryonic development to blastocyst stage. However, the clear transfer fetus which is expressing EGFP gene could not be obtained so far.
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