In vitro effects of tongue sole LPXRFa and kisspeptin on relative abundance of pituitary hormone mRNA and inhibitory action of LPXRFa on kisspeptin activation in the PKC pathway

Results of previous studies indicated the existence of LPXRFa, the piscine ortholog of gonadotropin-inhibitory hormone (GnIH), and kisspeptin (Kiss2) in tongue sole ( Cynoglossus semilaevis ), and that LPXRFa exerts an inhibitory effect on Kiss2 activation in the protein kinase A (PKA) pathway. The functions in the control of reproduction and whether LPXRFa antagonizes the action of Kiss2 by inhibiting the protein kinase C (PKC) pathway, however, are still unknown. In the present study, there was an initial investigation of the direct effects of LPXRFa and Kiss2 on relative abundance of pituitary hormone mRNA transcripts using a whole pituitary culture system. Results indicated that LPXRFa-1 specifically functioned to increase relative abundance of lhβ mRNA when there were comparisons with the control, without any effect on relative abundance of gh, gthα and fshβ mRNA. Treatment with LPXRFa-2 resulted in a reduction in relative abundance of gthα and lhβ mRNA, and did not alter relative abundance of fshβ mRNA. Treatment of LPXRFa-2 resulted in a greater relative abundance of gh mRNA. Treatment with Kiss2, however, resulted in an increase in relative abundance of gthα and fshβ mRNA transcripts, without altering relative abundances of gh and lhβ mRNA. Subsequently, there was valuation of the potential interaction between LPXRFa and kisspeptin in COS-7 cells transfected with the cognate receptors. Both LPXRFa-1 and LPXRFa-2 suppressed serum responsive element-dependent luciferase (SRE-luc) activity when compared to stimulation with Kiss2 alone, indicating an inhibitory effect of LPXRFa on kisspeptin activation on the PKC pathway. Overall, data from the present study provide novel evidence for differential actions of LPXRFa and kisspeptin on pituitary hormone synthesis as well as for the interaction between LPXRFa and kisspeptin systems in teleosts.