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Submitted on March 16, 2005
Accepted on June 27, 2005
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China; Graduate School, Chinese Academy of Sciences, Beijing 100080, China; Department of Veterinary Pathobiology, University of Missouri-Columbia, Columbia, MO 65211
* To whom correspondence should be addressed. E-mail: sunqy1{at}yahoo.com.
Mitogen-activated protein kinase (MAPK) plays important roles during meiotic maturation in mammalian oocytes, while the necessity of MAPK during meiotic resumption in porcine oocytes is still controversial. Here, by applying the method of ultracentrifugation to move the opaque lipid droplets to the edge of the oocyte therefore allowing clear visualization of porcine GVs, oocytes just before GVBD and those that had just undergone GVBD were selected for the assay of MAPK activation. Our results showed that phosphorylation of MAPK in oocytes occurred after GVBD in all three different culture models: spontaneous maturation model, inhibition-induction maturation model and normal maturation model. Moreover, we found that activation of MAPK in cumulus cells but not in oocytes was essential for GVBD in cumulus-enclosed oocytes. Then the crosstalk between cAMP and MAPK in cumulus cells was investigated by using cell-type specific phosphodiesterase (PDE) isoenzyme inhibitors. Our results showed that PDE3 subtype existed in oocytes whereas PDE4 subtype existed in cumulus cells. PDE3 inhibitor prevented meiotic resumption of oocytes, while PDE4 inhibitor enhanced the ability of FSH or forskolin to activate MAPK in cumulus cells. We propose that increased cAMP resulting from inhibition of PDE3 in oocytes blocks GVBD whereas increased cAMP resulting from inhibition of PDE4 activates MAPK pathway in cumulus cells, which is essential for GVBD induction.
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