MEK inhibitors block AICAR‐induced maturation in mouse oocytes by a MAPK‐independent mechanism

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Abstract
The present study was carried out to assess the possible role of mitogen‐activated protein kinase (MAPK) in the meiosis‐inducing action of the AMP‐activated protein kinase (AMPK) activator, 5‐aminoimidazole‐4‐carboxamide 1‐β‐ribofuranoside (AICAR). Cumulus cell‐enclosed oocytes (CEO) or denuded oocytes (DO) from immature, eCG‐primed mice were cultured 4 hr in Eagle's minimum essential medium containing dbcAMP plus increasing concentrations of AICAR or okadaic acid (OA). OA is a phosphatase inhibitor known to stimulate both meiotic maturation and MAPK activation and served as a positive control. Both OA and AICAR were potent inducers of meiotic resumption in mouse oocytes and brought about the phosphorylation (and thus, activation) of MAPK, but by different kinetics: MAPK phosphorylation preceded GVB in OA‐treated oocytes, while that resulting from AICAR treatment appeared only after GVB. The MEK inhibitors, PD98059 and U0126, blocked the meiotic resumption induced by AICAR but not that induced by OA. Although the MEK inhibitors suppressed MAPK phosphorylation in both OA‐ and AICAR‐treated oocytes, meiotic resumption was not causally linked to MAPK phosphorylation in either group. Furthermore, AICAR‐induced meiotic resumption in Mos‐null oocytes (which are unable to stimulate MAPK) was also abrogated by PD98059 treatment. A non‐specific effect of the MEK inhibitors on AICAR accessibility to the oocyte was discounted by showing that they failed to suppress either nucleoside uptake or AICAR‐stimulated phosphorylation of acetyl CoA carboxylase (ACC), a substrate of AMPK. The suppression of AICAR‐induced maturation by MEK inhibitors must, therefore, be occurring by actions unrelated to MEK stimulation of MAPK; consequently, it would be prudent to consider this possible non‐specific action of the inhibitors when they are used to block MAPK activation in mouse oocytes. Mol. Reprod. Dev. 70: 235–245, 2005.