Browse > Article

Effect of $Ca^{2+}$ Concentration in Fusion Medium on the Fusion, Nuclear Morphology and Development of Bovine Somatic Cell Nuclear Transfer Embryos  

조재원 (강원대학교 동물자원과학대학)
김정익 (강원대학교 동물자원과학대학)
박춘근 (강원대학교 동물자원과학대학)
양부근 (강원대학교 동물자원과학대학)
정희태 (강원대학교 동물자원과학대학)
Publication Information
Korean Journal of Animal Reproduction / v.26, no.1, 2002 , pp. 33-39 More about this Journal
Abstract
This study was conducted to investigate the effect of $Ca^{2+}$ concentration in fusion medium on the fusion, nuclear morphology and the development of bovine somatic cell nuclear transfer embryos. Bovine skin cells were transferred into an enucleated oocyte and fused with cytoplasm in the fusion medium containing with 0.05 to 1.0 mM Cacl$_2$. Nuclear transfer embryos were activated with a combination of A23187 and cycloheximide. Nuclear transfer embryos were fixed at 3 h after fusion or cultured for 7 ~8 days. Fusion rate was significantly (P＜0.01) increased by increasing the $Ca^{2+}$ concentrations in the fusion medium from 0.05 mM (56.6%) to 0.5 mM (50.1%) and 1.0 mM (84.3%). More than 80% of reconstituted embryos underwent premature chromosome condensation (PCC) with 0.05, 0.1 mM CaCl$_2$, whereas 54.5% and 59.3% of embryos formed pronucleus (PN) directly without PCC in the 0.5 and 1.0 mM CaCl$_2$, groups. Blastocyst formation rates were significantly (P＜0.05) different between 0.1 mM and 1.0 mM CaCl$_2$groups. From the present result, it is suggested that the elevated $Ca^{2+}$ concentrations in fusion medium can enhance the fusion and blastocyst formation rates of bovine nuclear transfer embryos.bryos.
Keywords
Somatic cell nuclear transfer; $Ca^{2+}$ concentration; Fusion rate; In vitro development; Bovine;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Aoyagi, Y. and Konish, M. 1994. Studies on development into blastocyst of in vitro matured and artificial bovine oocytes-mixed activation method by use of electric pulse, $Ca^{2+}$-ionophore and cycloheximide. J. Reprod. Dev. 40:5-11
2 Cheong, H. T., Park, K. W., Im, G. S., Lai, L., Sun, Q. Y., Day, B. N. and Prather, R. S. 2002. Effect of elevated $Ca^{2+}$ concentration in fusion/activation medium on the fusion and development of porcine fetal fibroblast nuclear transfer embryos. Mol. Reprod. Dev. in press
3 Lavoir, M. C., Rumph, N., Moens, A., King, W. A., Plante, Y., Tohnson, W. H., Ding, J. and Betteridge, K. J. 1997. Development of bovine nuclear transfer embryos made with oogonia. Biol. Reprod. 56:194-199   DOI   ScienceOn
4 Wells, D. N., Misica, P. M. and Tervit, H. R. 1999. Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells. Biol. Reprod. 60:996-1005   DOI   ScienceOn
5 Wilmut, I., Schnieke, A. E., McWhir, J., Kind, A. J. and Campbell, K. H. S. 1997. Viable off-spring derived from fetal and adult mammalian cells. Nature 385:810-813
6 최종엽, 권대진, 김정익, 박춘근, 양부근, 정희태. 2000. 세포의 휴면처리가 소 태아섬유아세포 유래 핵이식란의 핵상변화와 체외발육에 미치는 영향. 한국가축번식학회지 24:217-222   과학기술학회마을
7 Barnes, F. L., Collas, P., Powell, R., King, W. A., Westhusin, M. and Shepherd, D. 1993. In-fluence of recipient oocytes cell cycle stage on DNA synthesis, nuclear envelop breakdown, chromosome constitution, and development in nuclear transplant bovine embryos. Mol. Reprod. Dev. 36:33-41
8 Presicce, G. A. and Yang, X. 1993. Dynamics of activation of in vitro matured bovine follicu-lar oocytes following combined ethanol and cycloheximide. Mol. Reprod. Dev. 38:380-385
9 Wells, D. N., Misica, P. M., Day, A. M. and Tervit, H. R. 1997. Production of cloned lambs from an established embryonic cell line: A comparison between in vivo- and in vitro-ma-tured cytoplasts. Biol. Reprod. 57 : 385-393   DOI   ScienceOn
10 Collas, P., Fissore, R., Robl, J. M., Sullivan, E. J. and Barnes, F. L. 1993. Electrically induced calcium elevation, activation, and parthenogene-tic development of bovine oocytes. Mol. Reprod. Dev. 34:212-223   DOI   ScienceOn
11 First, N. L., Leibfried-Rutledge, M. L., Nor-they, D. L. and Nuttleman, P. R. 1992. Use of in vitro matured oocytes 24hr of age in bovine nuclear transfer. Theriogenology 37:211(abstr.)
12 Campbell, K. H. S., Loi, P., Otaegui, P. J. and Wilmut, I. 1996. Cell cycle co-ordination in embryo cloning by nuclear transfer. Rev. Reprod. 1:40-46
13 Sun, F. Z., Hoyland, J., Huang, X., Mason, W. and Moor, R. M. 1992. A comparison of intra-cellular changes in porcine eggs after fertiliza-tion and electroactivation. Development 115:947-956   PUBMED
14 Cheong, H. T., Takahashi, Y. and Kanagawa, H. 1993. Birth of mice after transplantation of early cell-cycle-stage embryonic nuclei into e-nucleated oocytes. Biol. Reprod. 48:958-963   DOI   ScienceOn
15 Cheong, H. T., Takahashi, Y. and Kanagawa, H. 1994. Relationship between nuclear remod-eling and subsequent development of mouse embryonic nuclei transferred to enucleated oo-cytes. Mol. Reprod. Dev. 37:138-145   DOI   ScienceOn
16 Westhusin, M. E., Levanduski, M. J., Scar-borough, R., Looney, C. R. and Bondioli, K. R. 1992. Viable embryos and normal calves after nuclear transfer into Hoechst stained enucleated demi-oocytes of cow. J. Reprod. Fertil. 95:475-480   DOI   PUBMED   ScienceOn