Protein kinase C (PKC) role in bovine oocyte maturation and early embryo development.
The aims of the present study were to determine the role of protein kinase C (PKC) on meiotic resumption and its effects on pronuclear formation and cleavage in the bovine. Oocytes were matured in the presence of 0, 1, 10 and 100 nM of phorbol 12-myristate 13-acetate (PMA), to evaluate the percentage of germinal vesicle breakdown. To study pronuclear formation and cleavage, oocytes were randomly distributed in four groups and matured in modified TCM-199 with LH and FSH (negative control); 10% of estrous cow serum (positive control); 100 nM of PMA (treatment); 100 nM of 4alpha-PDD (phorbol ester control). Oocytes were also matured in positive control medium, fertilized and transferred to KSOM with increasing concentrations of a PKC inhibitor. The protein profile and the presence of PKC at the end of maturation period were determined by SDS-PAGE followed by Silver Stain and Western blot, respectively. PMA stimulated meiotic resumption in a concentration-dependent manner. PKC stimulation during oocyte maturation caused an increase in pronuclear formation and did not cause parthenogenetic activation. Inhibitor of PKC (MyrPKC) inhibited cleavage in a dose-dependent and irreversible manner. A protein band around 74 kDa was not detected in PMA-treated oocytes and PKC was not detected by Western blot at the end of the maturation period. In conclusion, meiotic resumption was accelerated and the rate of oocytes with two pronuclei was increased when PKC was activated during oocyte maturation. Moreover, cleavage was inhibited in the presence of PMA.
Mondadori
R G
RG
Animal Reproduction Laboratory, Department of Veterinary Medicine, UPIS-Faculdades Integradas, SEPS 712/912, Brasilia, DF 70390-125, Brazil. mondadori@upis.br
Neves
J P
JP
Gonçalves
P B D
PB
D000818:Animals; D015153:Blotting, Western; D002417:Cattle / Q000196:embryology*; D002478:Cells, Cultured; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D020869:Gene Expression Profiling; D008540:Meiosis / Q000502:physiology; D009865:Oocytes / Q000166:cytology / Q000201:enzymology*; D010703:Phorbol Esters / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction
D010703:Phorbol Esters; C008874:phorbol-12,13-diacetate; D011493:Protein Kinase C