Pituitary Adenylate Cyclase-activating Polypeptide and Cyclic Adenosine 3′,5′-Monophosphate Stimulate the Promoter Activity of the Rat Gonadotropin-releasing Hormone Receptor Gene via a Bipartite Response Element in Gonadotrope-derived Cells

Abstract

Specific type I receptors for pituitary adenylate cyclase-activating polypeptide (PACAP) are present in gonadotrope cells of the anterior pituitary gland. By transient transfection of mouse gonadotrope-derived αT3-1 cells, which are direct targets for PACAP and express gonadotropin-releasing hormone receptor (GnRH-R), a marker of the gonadotrope lineage, we provide the first evidence that PACAP stimulates rat GnRH-R gene promoter activity. The EC 50 of this stimulation is compatible with a mediation via activation of the cyclic AMP-dependent signaling pathway and, consistently, co-transfection of an expression vector expressing the protein kinase A inhibitor causes reduction in PACAP as well as cholera toxin-stimulated promoter activity. Deletion and mutational analyses indicate that PACAP activation necessitates a bipartite response element that consists of a first region (-272/-237) termed PACAP response element (PARE) I that includes a steroidogenic factor-1 (SF-1)-binding site and a second region (-136/-101) referred to as PARE II that contains an imperfect cyclic AMP response element. Gel shift experiments indicate the specific binding of the SF-1 and a potential SF-1-interacting factor to PARE I while a protein immunologically related to the cyclic AMP response element-binding protein interacts with PARE II. These findings suggest that PACAP might regulate the GnRH-R gene at the transcriptional level, providing novel insights into the regulation of pituitary-specific genes by hypothalamic hypophysiotropic signals.