Kisspeptin induces expression of gonadotropin-releasing hormone receptor in GnRH-producing GT1–7 cells overexpressing G protein-coupled receptor 54

• We found that kisspeptin, but not GnRH increases GnRHR promoter activity in GnRH-producing GT1–7 cells.
• Kisspeptin stimulates both ERK and cAMP/PKA signaling pathways, but GnRH increases only ERK pathway.
• When GT1–7 cells were stimulated by GnRH in the presence of cAMP, GnRHR promoters were significantly increased.
• Kisspeptin is a potent stimulator of GnRHR expression in GT1–7 cells in association with ERK and the cAMP pathways.

Kisspeptin signaling through its receptor is crucial for many reproductive functions. However, the molecular mechanisms and biomedical significance of the regulation of GnRH neurons by kisspeptin have not been adequately elucidated. In the present study, we found that kisspeptin increases GnRH receptor (GnRHR) expression in a GnRH-producing cell line (GT1–7). Because cellular activity of G protein-coupled receptor 54 (GPR54) and GnRHR was limited in GT1–7 cells, we overexpressed these receptors to clarify receptor function. Using luciferase reporter constructs, the activity of both the serum response element (Sre) promoter, a target for extracellular signal-regulated kinase (ERK), and the cyclic AMP (cAMP) response element (Cre) promoter were increased by kisspeptin. Although GnRH increased Sre promoter activity, the Cre promoter was not significantly activated by GnRH. Kisspeptin, but not GnRH, increased cAMP accumulation in these cells. Kisspeptin also increased the transcriptional activity of GnRHR; however, the effect of GnRH on the GnRHR promoter was limited and not significant. Transfection of GT1–7 cells with constitutively active MEK kinase (MEKK) and protein kinase A (PKA) increased GnRHR expression. In addition, GnRHR expression was further increased by co-overexpression of MEKK and PKA. The Cre promoter, but not the Sre promoter, was also further activated by co-overexpression of MEKK and PKA. GnRH significantly increased the activity of the GnRHR promoter in the presence of cAMP. The present findings suggest that kisspeptin is a potent stimulator of GnRHR expression in GnRH-producing neurons in association with ERK and the cAMP/PKA pathways.