Stimulation of δ subunit-containing GABAA receptor by DS1 increases GnRH receptor expression but reduces GnRH mRNA expression in GnRH-producing GT1-7 cells

Abstract

Acting via ionotropic GABAA receptors, the neurotransmitter γ-aminobutyric acid (GABA) is an important modulator of gonadotropin-releasing hormone (GnRH) neurons. In the present study, we examined the effect of DS1, a GABAA α4β3δ receptor agonist, on a strain of mouse hypothalamic immortalized GnRH neuronal cells, the GT1-7 cell line. DS1 increased the activities of serum-response element (SRE) and cAMP-response element (CRE) promoters, which reflect the activities of extracellular signal-regulated kinase and cAMP/protein kinase A (PKA) pathways, respectively. In G protein-coupled receptor 54 (GPR54)-overexpressing GT1-7 cells, both DS1 and kisspeptin-10 stimulated SRE promoter activity, and combined treatment with DS1 and kisspeptin further increased SRE promoter activity compared with DS1 or kisspeptin alone. Pituitary adenylate cyclase-activating polypeptide (PACAP) increased CRE promoter activity in PACAP type I receptor-overexpressing GT1-7 cells, with an effect similar to that of DS1 alone, and combined stimulation with PACAP and DS1 potentiated their individual effects. DS1 stimulated the transcriptional activity of GnRH receptor, and DS1 induced GnRH receptor mRNA and protein expression. PACAP-increased GnRH receptor expression was enhanced in the presence of DS1. However, DS1 significantly inhibited the basal expression of GnRH mRNA in GT1-7 cells. Our current observations suggest that DS1 exerts its stimulatory effect on the intracellular signal transduction system via GABAA α4β3δ receptors in GnRH-producing neurons. Stimulation with DS1 increased the expression of GnRH receptor but decreased the basal expression of GnRH mRNA.