Activation of Akt (protein kinase B) stimulates metaphase I to metaphase II transition in bovine oocytes.
In somatic cells, the serine/threonine kinase Akt (or protein kinase B) was shown to contribute to processes linked to cellular growth, cell survival and cell cycle regulation. In contrast to these findings, the function of Akt during the meiosis of mammalian oocytes remains to be investigated. We analysed the phosphorylation pattern and the activity of Akt during meiotic maturation (transition from prophase I to metaphase II) of bovine oocytes. The oocytes were matured in vitro (IVM) for 0, 10 and 24 h to reach the germinal vesicle (GV), metaphase I (M I) and metaphase II (M II) stages respectively. The abundance and phosphorylation pattern of Akt was revealed by Western blotting using total Akt or phosphoso-Akt-specific antibodies. The activity of this particular kinase was determined by an in vitro kinase assay. Furthermore, functional properties were analysed by cultivating oocytes in the presence of the Akt inhibitor SH6. The results showed that the overall abundance of Akt did not change significantly during IVM. On the other hand, Akt became phosphorylated at Thr 308 and Ser 473, reaching its maximum at the M I phase. In the GV and M II stages, only low basal phosphorylation levels were observed on both sides. This phosphorylation profile corresponded strictly to the activity of the kinase. The cultivation of oocytes in the presence of the phosphatidylinositol analogue SH6 for 24 h showed that, with higher concentrations, up to 65% of the oocytes were arrested in the M I stage. This result indicated that Akt is involved in the M I/M II transition during the meiotic maturation of bovine oocytes. The physiological aspects of the Akt function will be discussed.
Tomek
Wolfgang
W
Unit of Reproductive Biology, FBN Research Institute for the Biology of Farm Animals, Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany. tomek@fbn-dummerstorf.de
Smiljakovic
Tatjana
T
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002417:Cattle; D018797:Cell Cycle Proteins / Q000032:analysis; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005260:Female; D020935:MAP Kinase Signaling System / Q000502:physiology; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008677:Metaphase / Q000502:physiology*; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D010716:Phosphatidylinositols / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000032:analysis; D051057:Proto-Oncogene Proteins c-akt / Q000032:analysis / Q000378:metabolism*
D018797:Cell Cycle Proteins; D010716:Phosphatidylinositols; D011494:Protein Kinases; C102333:histone H1 kinase; D051057:Proto-Oncogene Proteins c-akt; D016200:Maturation-Promoting Factor