Roles of cAMP in regulation of both MAP kinase and p34cdc2 kinase activity during meiotic progression, especially beyond the MI stage

Abstract

Time‐dependent changes in the level of adenosine cyclic AMP (cAMP) in porcine oocytes during meiotic progression from the germinal vesicle stage (GV stage) to the metaphase II stage (MII stage) were examined using reversed‐phase HPLC with UV detection. The concentration of cAMP in oocytes reached a peak at 8 hr of cultivation of cumulus–oocyte complexes (COCs), but it was dramatically decreased after 12‐hr cultivation. After a 28‐hr cultivation period, the level of cAMP in the oocytes had significantly reduced further, and the basal level of cAMP was observed in oocytes cultured at 32 hr and for up to 48 hr. When phosphatidylinositol 3‐kinase (PI 3‐kinase) or protein kinase C (PKC) in cumulus cells [which were required for meiotic progression to the MII stage in porcine oocytes (Shimada and Terada, 2001: Biol Reprod 64:1106–1114)] was suppressed by each specific inhibitor following initial 24‐hr cultivation of COCs, cAMP level in the oocytes was significantly increased. After 24‐hr cultivation in the maturation medium, COCs, which were cultured for an additional 24 hr in the presence of either forskolin or 3‐isobutyl‐1‐methylxanthine (IBMX), exhibited a significant increase in the oocyte cAMP level to the similar level of that in oocytes cultured with PI 3‐kinase inhibitor or PKC inhibitor, and the addition of each agent significantly suppressed meiotic progression from the MI to the MII stage and the activity of mitogen‐activated protein kinase (MAPK) and p34^cdc2 kinase. These results demonstrated that when transported into oocytes from the cumulus cells via gap junctions, cAMP plays an important role not only in meiotic resumption, but also in the regulation of meiotic progression beyond the MI stage in porcine oocytes. Mol. Reprod. Dev. 62: 124‐131, 2002.