JARB Journal of Animal Reproduction and Biotehnology

OPEN ACCESS pISSN: 2671-4639
eISSN: 2671-4663

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Journal of Embryo Transfer 2002; 17(1): 13-21

Published online April 30, 2002

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Development of Bovine Embryos Produced by Intracytoplasmic Sperm Injection (ICSI)

Ock, S.A., Kwack, D.O., Cho, S.R., Cho, S.K., Yeao, E.H., Yoo, J.G., Lee, Y.R., Lee, H.J., Choe, S.Y., Rho, G.J.

Apllied Animal Science, Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University

Abstract

Intracytoplasmic Sperm Injection (ICSI) has been widely used fur both human infertility and basic research. However, the high incidence of chromosomal abnormality is severe problem in cattle. Various oocyte activation stimuli, therefore, were compared by assessment of developmental capacity and chromosome analysis. Motile sperm selected by Percoll-density gradient were treated with 5 mM dithiothreitol (DTT) and injected into an oocyte matured fur 24 h. Eggs were then allocated into 5 treatment groups. Group 1 (control), sperm injection was performed without any further activation stimuli to the oocytes. Group 2 (handled control), sham injection was performed without sperm. In Group 3, oocytes exposed to 5 (M ionomycin for 5 min at 39(C. Group 4. ionomycine + 1.9 mM demethylaminopurine (DMAP, 3 h) and Group 5, ionomycine + 3 h culture in Ml99 + DMAP. Cleavage and the later development rate in Groups 1, 2 and 3 were significantly (P<0.05) lower than those in Groups 4 and 5. The incidence of chromosomal abnormality in the embryos treated directly with DMAP after ionomycine was relatively higher than in the embryo of Group 3 h, delayed DMAP treatment. From this results DMAP caused to be arrested the release of the 2nd polar body, resulting in changes of chromosomal pattern. Therefore, the time interval between ionomycin and DMAP is a crucial role in bovine ICSI.

Keywords: ploidy, activation, ICSI, bovine oocyte

Article

Journal of Embryo Transfer 2002; 17(1): 13-21

Published online April 30, 2002

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Development of Bovine Embryos Produced by Intracytoplasmic Sperm Injection (ICSI)

Ock, S.A., Kwack, D.O., Cho, S.R., Cho, S.K., Yeao, E.H., Yoo, J.G., Lee, Y.R., Lee, H.J., Choe, S.Y., Rho, G.J.

Apllied Animal Science, Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University

Abstract

Intracytoplasmic Sperm Injection (ICSI) has been widely used fur both human infertility and basic research. However, the high incidence of chromosomal abnormality is severe problem in cattle. Various oocyte activation stimuli, therefore, were compared by assessment of developmental capacity and chromosome analysis. Motile sperm selected by Percoll-density gradient were treated with 5 mM dithiothreitol (DTT) and injected into an oocyte matured fur 24 h. Eggs were then allocated into 5 treatment groups. Group 1 (control), sperm injection was performed without any further activation stimuli to the oocytes. Group 2 (handled control), sham injection was performed without sperm. In Group 3, oocytes exposed to 5 (M ionomycin for 5 min at 39(C. Group 4. ionomycine + 1.9 mM demethylaminopurine (DMAP, 3 h) and Group 5, ionomycine + 3 h culture in Ml99 + DMAP. Cleavage and the later development rate in Groups 1, 2 and 3 were significantly (P<0.05) lower than those in Groups 4 and 5. The incidence of chromosomal abnormality in the embryos treated directly with DMAP after ionomycine was relatively higher than in the embryo of Group 3 h, delayed DMAP treatment. From this results DMAP caused to be arrested the release of the 2nd polar body, resulting in changes of chromosomal pattern. Therefore, the time interval between ionomycin and DMAP is a crucial role in bovine ICSI.

Keywords: ploidy, activation, ICSI, bovine oocyte