1999 Fiscal Year Final Research Report Summary
Study on Activation of Oocytes Which Are Injected With a Round Spermatid
| Project/Area Number |
10671549
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | the University of Tokushima |
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Principal Investigator |
YAMANO Shuji The University of Tokushima, University Hospital, Associate Professor, 医学部・附属病院, 助教授 (30166772)
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| Co-Investigator(Kenkyū-buntansha) |
KOMATSU Junko The University of Tokushima, University Hospital, medical staff, 医学部・附属病院, 医員(臨床)
YAMASITA Mizuho The University of Tokushima, University Hospital, medical staff, 医学部・附属病院, 医員(臨床)
NAKAGAWA Koji The University of Tokushima, University Hospital, Research Associate, 医学部・附属病院, 助手 (40314869)
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| Project Period (FY) |
1998 – 1999
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| Keywords | fertilization / oocyte activation / parthenogenone / MPF / MAPK |
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| Research Abstract |
Recent reports suggest that when unfertilized oocytes with a spermatozoon after intracytoplasmic sperm injection (ICSI) are properly activated, the activated oocytes develop normally like fertilized oocytes. The aim of the study is to develop the activation method which is able to produce embryos with normal karyotype from unfertilized oocytes with a spermatozoon, the activation method which produces one pronucleus with extrusion of the second polar body in oocytes without a spermatozoon is necessary.
1) When human unfertilized oocytes 22 h after insemination were treated with 5 M Calcium ionophore for 5 min and incubated in HTF medium with 10 g/ml puromycin for 5 h, the activation rate was about 90% and the proportion of parthenogenones displaying one pronucleus (PN) with extrusion of the second polar body (PB) was about 80%.
2) When mouse ovulated oocytes were treated with 5 M Calcium ionophore for 5 min and incubated in HTF medium with 10 g/ml puromycin for 4 h, the activation rate and the proportion of parthenogenones displaying one PN with extrusion of the second PB were similar to those of human aged oocytes treated with the same method.
3) When human unfertilized oocytes following ICSI were activated by the method, two PN were formed with extrusion of the second PB in only 30% of the parthenogenones and 64% of the parthenogenones with 2PN2PB cleaved. Four clcaved parthenogenones showed normal karyotype.
4) When maturation promoting facor (MPF) and mitogen activated protein kinase (MAPK) in mouse oocytes treated with the method were measured. MPF significantly decreased first in a time-dependent manner, and sequentially MAPK decreased.
These findings suggest that the activation method decreased MPF and MAPK like fertilized oocytes, and extrudes the second PB and forms a female PN. However, puromycin may suppress the formation of a male PN.
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[Publications] Nakagawa, K., Yamano, S., Kamada, M., Hinokio, K., Maegawa, M., Aono, T.: "Quality of embryo does not affect the implantation rate of in vitro fertilization embryo transfer in infertile women with antisperm antibody"Fertil Steril. 72. 1055-1060 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Nakasaka, H., Yamano, S., Hinokio, K., Nakagawa, K., Yoshizawa, M., Aono, T.: "Effective activation method with A23187 and puromycin to produce haploid parthenogenones from freshly ovulated mouse oocytes"Zygote. 8. 203-208 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Quan, S., Yamano, S., Nakasaka, H., Hinokio, K., Nakagawa, K., Aono, T.: "Effects of preservation of mouse spermatozoa in electrolite-free solution at 4℃ on the outcome of mouse, in vitro fertilization"J Assist Reprod Genet. 17. 384-388 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Takagawa, K., Yamano, S., Nakasaka, H., Hinokio, K., Yoshizawa, M., Aono, T.: "A combination of Ca ionophore and puromycin effectively produces human parthenogenones with noe haploid pronucleus"Zygote. (in press).
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[Publications] Takikawa M., Kamada M., Maegawa M., Yamano S., Irahara M., Aono T., Futaki S., Ohmoto Y., Koide S.S.: "Evaluation of two sperm antigens, rSMP-B and YWK-II, as targets for immuno-contraception"Zygote. (in press).
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「研究成果報告書概要(欧文)」より
