Effect of activation time on the nuclear remodeling and in vitro development of nuclear transfer embryos derived from bovine somatic cells.
This study was conducted to investigate the effect of recipient activation time on the chromatin structure and development of bovine nuclear transfer embryos. Serum-starved skin cells were electrofused to enucleated oocytes, activated 1-5 hr after fusion, and cultured in vitro. Some fused eggs were fixed at each time point after fusion without activation, or 3 or 7 hr after activation. Some nocodazole treated zygotes were fixed to analyze their chromosome constitutions. The proportion of eggs with a morphologically normal premature chromosome condensation (PCC) state increased 1-2 hr after fusion. Whereas eggs with elongated chromosome plate increased as activation time was prolonged to 3 hr, and 5 hr after fusion, 58.1% of eggs showed more than two scattered chromosome sets. The proportion of eggs with a single chromatin mass (40.6-56.7%) significantly increased when eggs were activated within 2.5 hr after fusion (P < 0.05). Only 23.3% of reconstituted embryos activated 5 hr after fusion formed one pronucleus-like structure (PN), whereas, 64.5-78.3% of embryos activated 1-2.5 hr after fusion formed one PN. The proportion of embryos with normal chromosome constitutions decreased as activation time was prolonged. Development rates to the blastocyst stage were higher in eggs activated within 2 hr after fusion (17.3-21.7%) compared to those of others (0-8.6%, P < 0.05). The result of the present study suggests that activation time can affect the chromatin structure and in vitro development of bovine nuclear transfer embryos.
Choi
Jong-Yeob
JY
Faculty of Animal Resource, College of Animal Resource Science, Kangwon National University, Chuncheon, Korea.
Kim
Choung-Ik
CI
Park
Choon-Keun
CK
Yang
Boo-Keun
BK
Cheong
Hee-Tae
HT
D000818:Animals; D030801:Animals, Genetically Modified; D002417:Cattle; D002467:Cell Nucleus / Q000502:physiology*; D002843:Chromatin / Q000502:physiology*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology*; D004622:Embryo, Mammalian / Q000502:physiology*; D053652:Nuclear Transfer Techniques; D013997:Time Factors
D002843:Chromatin