Effect of roscovitine, a cdk1 inhibitor, and of the presence of oocyte on bovine cumulus cell expansion and cyclooxygenase-2 expression.
Oocyte maturation is accompanied by differentiation of surrounding cumulus cells. These cells produce hyaluronic acid (HA) and its storage in intercellular spaces results in expansion of the cells. The cumulus cells also accumulate cyclooxygenase-2 (cox-2) during maturation. Both expansion and cox-2 storage are regulated by FSH and EGF. The aim of this study was to determine whether oocyte meiotic resumption is involved in the regulation of cumulus differentiation or not. We investigated the effects of roscovitine, a reversible inhibitor of meiosis resumption of cattle oocytes on EGF induced expansion and cox-2 expression at the transcript and protein levels respectively (RT-PCR and Western blot), in cumulus oocyte complexes (COCs) and cumulus complexes alone (CCs). EGF induced expansion and cox-2 expression in both COCs and CCs. These effects were prevented by roscovitine, whether in the presence or in the absence of oocyte. However, the oocyte was essential for the reversibility of inhibition by roscovitine. In conclusion, our results indicate that i) oocyte secreted-factors are not essential for cumulus expansion, and ii) roscovitine mediated inhibition of meiotic resumption also respects the functionality of the surrounding somatic cells.
Vigneron
C
C
Institut National de la Recherche Agronomique, Unité de Physiologie de la Reproduction et des Comportements, Nouzilly, France.
Nuttinck
F
F
Perreau
C
C
Reinaud
P
P
Charpigny
G
G
Mermillod
P
P
D000818:Animals; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors; D002417:Cattle; D051546:Cyclooxygenase 2; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D006080:Ovarian Follicle / Q000187:drug effects*; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D011687:Purines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D000077546:Roscovitine
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D011687:Purines; D012333:RNA, Messenger; D000077546:Roscovitine; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D016203:CDC2 Protein Kinase