Developmental and protein kinase-dependent regulation of ovarian connexin mRNA and oocyte maturational competence in Atlantic croaker

The acquisition of oocyte maturational competence (OMC) in ovarian follicles of Atlantic croaker is associated with increased gap junction (GJ) contacts and increased levels of ovarian connexin (Cx) 32.2 mRNA. However, the developmental control of ovarian Cx gene expression and the mechanisms of OMC acquisition are unknown. Ovarian Cx32.2 and Cx32.7 mRNA levels were determined in fish with gonadosomatic indices (GSI; gonad weight-to-body weight ratio) ranging from 0.1 to 13%. The mRNA level for both Cx increased from a low level in previtellogenic ovaries (GSI, <1%) to a peak level during the midstage of ovarian growth (GSI, 6-7%). Levels of Cx32.2 mRNA, but not Cx32.7 mRNA, declined markedly during late ovarian vitellogenic growth (GSI, 7-13%), and increased again upon stimulation of OMC by human chorionic gonadotropin (hCG). These changes in ovarian Cx32.2 mRNA seem to parallel previously reported changes in the incidence of oocyte-granulosa cell GJ during follicular growth and early maturation. In vitro treatment with hCG and protein kinase A (PKA) activators (dbcAMP and forskolin) induced ovarian Cx32.2 mRNA levels and OMC. The effects of hCG were blocked by PKA inhibitors (H89, H7). Protein kinase C (PKC) inhibitors (GF 109207X) had little effect on hCG-induced Cx32.2 mRNA or OMC, whereas PKC activators (PMA) blocked both events. There was no association between changes in Cx32.7 mRNA levels and OMC status in these experiments. In conclusion, changes in Cx32.2 gene expression seem to be involved in the regulation of oocyte-granulosa cell GJ during growth and differentiation of the croaker ovarian follicle. Also, the stimulation of OMC and Cx32.2 mRNA levels by hCG is mediated by PKA-dependent pathways and antagonized by PKC-dependent mechanisms.