Induction of c-fos and c-jun messenger ribonucleic acid expression by prostaglandin F2 alpha is mediated by a protein kinase C-dependent extracellular signal-regulated kinase mitogen-activated protein kinase pathway in bovine luteal cells

PGF2 alpha triggers the demise of the corpus luteum whereby progesterone sy nthesis is inhibited, the luteal structure regresses, and the estrus cycle resumes. Upon binding to its heterotrimeric G-protein-coupled receptors, PG F2 alpha initiates the phospholipase C/diacylglycerol and inositol-1,4,5-tr isphosphate/Ca2+-protein kinase C (PKC) signaling pathway. More recently, w e have demonstrated that PGF2 alpha activates extracellular signal-regulate d kinase (ERK) mitogen-activated protein (MAP) kinase signaling through a R af-dependent mechanism in bovine luteal cells. However, the relationship be tween PKC and ERK activation in PGF2a signaling has not been clearly define d. Moreover, the signaling pathway that PGF2 alpha uses to regulate gene ex pression is unknown. In this report, primary cultures of bovine luteal cell s were used to address the role of PKC in ERK activation and the signaling pathway for induction of c-fos and c-jun messenger RNA (mRNA) expression in response to PGF2 alpha. By using a PKC inhibitor and a PKC-deficient lutea l cell model, we observed that phorbol ester-responsive isoforms of PKC wer e required for ERK phosphorylation and activation by PGF2 alpha (1 muM) or phorbol 12-myristate 13-acetate (PMA) (20 nM). In PGF2 alpha- and PMA-treat ed cells, active ERK MAP kinase was localized in the nucleus. PGF2 alpha -i nduced ERK phosphorylation was dose-dependently inhibited by the MEK1 inhib itor PD098059 (1-50 muM). The expression of c-fos and c-jun mRNA in luteal cells was markedly increased by treatment with PGF2 alpha (1 muM) or PMA (2 0 nM) for 30 min. We also observed that activation of ERK MAP kinase was re quired for the expression of c-fos and c-jun mRNA in response to PGF2 alpha and PMA because it was abrogated by blocking the ERK pathway with PD098059 . In addition, PGF2 alpha and PMA-induced c-fos and c-jun mRNA expression w as abolished in the PKC-deficient cells. Taken together, our data demonstra te that a PKC-dependent ERK MAP kinase pathway mediates the expression of c -fos and c-jun mRNA in PGF2 alpha -treated bovine luteal cells.