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Animal Reproduction and Physiology
Asian-Australasian Journal of Animal Sciences 1999;12(1): 22-27.
DOI: https://doi.org/10.5713/ajas.1999.22    Published online February 1, 1999.
Effect of Cycloheximide on Bovine Oocyte Nuclear Progression and Sperm Head Transformation after Fertilization In Vitro
L. Liu, H. W. Zhang, J. F. Qian, N. Fujihara
Abstract
Bovine oocytes with compact and complete cumulus cells were cultured in 6 groups for up to 24h in TCM199 buffered with 25 mmol/1 HEPES and supplemented with 10% FCS, 1 mg/ml 17 棺-Estradiol, 20 IU/ml hCG. Half of the oocytes at each group cultured in the presence of 25 關g/ml cycloheximide at different times during maturation (0, 6, 12, 18, 20, 22 h) were fixed at 24 h of maturation to examine the nuclear progression. The rests of them were inseminated with frozen-thawed spermatozoa in medium BO with 10 mg/ml BSA and 10 mg/ml heparin and fixed after additional 18-20 h culture to evaluate the sperm head transformation. When a protein synthesis inhibitor was added at the onset of the maturation., the oocytes were prevented to proceed GVBD. A few of the oocytes (16%) were able to be penetrated and sperm head decondensation was inhibited either. Addition of cycloheximide after 6-12 h of culture resulted in an increasing percentage of GVBCD (more than 80%), but the oocytes became arrested in M-I (69.2%). More than half of the oocytes was penetrated with a decondensing sperm head. Formation of male pronucleus was first obtained at 12 h of culture in the presence of cycloheximide. When cycloheximide was added from 18 h of culture onwards, nuclear progression to M-II was increasingly restored (80.4-85.5%). The proportion of male and female pronuclear formation increased from 17.9% to 46.2%. It is concluded that protein synthesis is necessary not only for GVBD and development from M-I to M-II, but also for sperm head decendensation and male pronuclear formation in bovine oocytes.
Keywords: Protein Synthesis; Bovine Oocytes; Maturation; Sperm Nucleus
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