Protein phosphorylation patterns during in vitro maturation of the goat oocyte.
Protein phosphorylation patterns were studied by radiolabelling goat cumulus oocyte complexes with [32P]orthophosphate for various periods of time. The radiolabelled denuded oocytes were assessed for nuclear status and were used individually for gel electrophoresis. This study demonstrated that specific changes in protein phosphorylations were programmed during goat oocyte maturation. One of the most prominent changes was a general increase in the phosphorylation rate at germinal vesicle breakdown (GVBD). From 8 hr of culture, dominant phosphoprotein bands with apparent molecular weights of 27, 31, 40, and 50 kD were observed; they remained at this level until the metaphase II stage. In the molecular weight range of 65-80 kD, the protein phosphorylation pattern exhibited characteristic differences, with a complex series of phosphoproteins appearing and disappearing, duration maturation. Addition of 6-dimethylaminopurine (6-DMAP) at the onset of culture blocked the maturation process after GVBD and induced a dramatic condensation of chromatin. When added at different times after GVBD, 6-DMAP invariably induced chromosome condensation. This inhibition was partly reversible; i.e., after removal of the drug, oocytes were able to progress only until metaphase I.
Gall
L
L
Unité de Biologie de la Fécondation, Station de Physiologie Animale, INRA, Jouy-en-Josas, France.
Le Gal
F
F
De Smedt
V
V
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D004527:Egg Proteins / Q000378:metabolism*; D005260:Female; D006041:Goats; D066298:In Vitro Techniques; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation
D004527:Egg Proteins; D010710:Phosphates; C001020:N(6),N(6)-dimethyladenine; D000225:Adenine