The Gα-provoked induction of Akr1c18 in murine luteal cells is mediated by phospholipase C.

Towards the end of gestation prostaglandin F2α (PGF2α) stimulates the expression of Akr1c18 in the murine corpus luteum. Akr1c18 codes for 20α-hydroxysteroid dehydrogenase, an enzyme that precipitates parturition by catabolizing progesterone. Previous results from our laboratory have shown that this effect of PGF2α is mediated by the activation of Gα, but the downstream effector(s) of Gα that elicit the increase in Akr1c18 expression have not been identified. The physiological effects of Gα are mediated by its ability to interact with phospholipase Cβ, p63RhoGEF, and PKCζ. In the experiments described herein we used biochemical and pharmacological approaches, as well as adenoviral-mediated expression of a constitutively active form of Gα and mutants thereof, to examine the role of each of these effectors as potential mediators of the increased expression of luteal Akr1c18. By measuring the effects of PGF2α on the activation of RhoA (activated by p63RhoGEF) and the effects of activators and inhibitors of RhoA on the PGF2α-induced expression of luteal Akr1c18, we determined that RhoA is neither activated by PGF2α or involved in the PGF2α-induced expression of luteal Akr1c18. The potential involvement of PKCζ was ruled out by the inability of a mutant of a constitutively active Gα that prevents PKCζ binding to block the increased expression of Akr1c18. Furthermore, PGF2α does not increase the phosphorylation of ERK-5, the only known downstream target of PKCζ. On the other hand, three different mutants of a constitutively active Gα that prevent phospholipase C activation blocked the induction of luteal Akr1c18. We conclude that the induction of luteal Akr1c18 by Gα is mediated by the activation of phospholipase C.