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Pituitary Adenylate Cyclase-Activating Peptide Stimulates Acute Progesterone Production in Rat Granulosa/Lutein Cells via Two Receptor Subtypes¹

^a Department of Clinical Biochemistry, University of Copenhagen, Bispebjerg Hospital, DK-2400 Copenhagen NV, Denmark

ABSTRACT

Pituitary adenylate cyclase-activating peptide (PACAP) is transiently expressed in ovarian granulosa/lutein cells from eCG/hCG-treated rats, and in vitro immunoneutralization of endogenously released PACAP inhibits acute progesterone secretion and subsequent luteinization in such cells. This suggests that PACAP mediates locally some of the effects of the LH surge, but the putative PACAP receptor(s) involved in such an auto or paracrine activity is presently unknown. Reverse-transcription polymerase chain reaction with specific primers to the three cloned PACAP-binding receptors called PAC₁, VPAC₁, and VPAC₂ demonstrated both PAC₁ and VPAC₂ mRNA in extracts from preovulatory follicular cells. Radioligand-binding assays revealed the presence of high-affinity binding sites with characteristics of these two receptors on the intact cells, and autoradiography demonstrated that the binding was restricted to a minor proportion of the follicular cells as well as the oocytes. Pituitary adenylate cyclase-activating peptide and vasoactive intestinal peptide (VIP) dose-dependently stimulated cAMP accumulation and acute progesterone accumulation. Forskolin and db-cAMP also stimulated acute progesterone accumulation, and the protein kinase A inhibitor H89 dose-dependently inhibited peptide induced acute progesterone accumulation, suggesting involvement of cAMP and the protein kinase A pathway in the process. In conclusion, two of the three PACAP binding receptors are present on preovulatory follicular cells and are involved in the effects of PACAP on acute progesterone production. The data provide further evidence to establish PACAP as an auto- or paracrine regulator of LH-induced acute progesterone production in rat preovulatory follicles.

FOOTNOTES

First decision: 2 November 2000.

¹ The study was supported by the Danish Biotechnology Center for Cellular Communication.

² Correspondence: Søren Gräs, Department of Clinical Biochemistry, University of Copenhagen, Bispebjerg Hospital, Bispebjerg Bakke 23, DK-2400 Copenhagen NV, Denmark. FAX: 45 35 31 39 55; s.gras{at}dadlnet.dk

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