PMID: 3760756Jun 1, 1986Paper

Protein requirements for germinal vesicle breakdown in ovine oocytes

Journal of Embryology and Experimental Morphology
R M Moor, I M Crosby

Abstract

The regulation of the cell cycle during the transition from prophase to metaphase I was studied by analysing protein changes and introducing protein blocks during the transition phase. The results show that the progression to metaphase in ovine oocytes is totally dependent on new protein synthesis. By delaying the addition of the inhibitor, cycloheximide, for progressively longer periods after the resumption of meiosis it was established that the required synthesis occurs in the 1-2 h immediately preceding germinal vesicle breakdown (GVBD). The action of cycloheximide was fully reversible: removal of the drug resulted in GVBD between 3 and 4 h later. The synthesis and modification of proteins during these first few hours of maturation were studied by short-term radiolabelling of oocytes with [35S]methionine and [32P]phosphate followed by rapid assessment of their precise nuclear configuration. Changes in phosphorylation of two polypeptides were detected 4-5 h after the beginning of culture, but these changes were not dependent upon protein synthesis. The earliest change in synthesis was the appearance of a new polypeptide 6-8 h after explantation, immediately before GVBD. This polypeptide (Mr 47 X 10(3), pI 5.8) was not signifi...Continue Reading

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