A novel inhibitory gonadotropin-releasing hormone-related neuropeptide in the ascidian, Ciona intestinalis

The gonadotropin-releasing hormone (GnRH) family peptides are most widely distributed neuropeptides and/or neurophysial hormones. GnRH is involved in diverse neuroendocrine, paracrine, autocrine, and neurotransmitter/neuromodulatory functions in the central and peripheral nervous system as well as peripheral tissues. In the present study, we show the identification of a novel GnRH-related peptide, Ci-GnRH-X, in the ascidian, Ciona intestinalis. Intriguingly, Ci-GnRH-X possesses a unique primary sequence consisting of 16 amino acids, although typical GnRH family peptides are composed of 10 amino acids. On the other hand, Ci-GnRH-X shares the GnRH consensus motifs, including the N-terminal pQHWS (‘pQ’ indicates a pyro-glutamic acid) and C-terminal Gly-amide. Reverse transcription (RT)-PCR analysis shows that the Ci-GnRH-X gene is expressed exclusively in the central nervous system. Moreover, in situ hybridization demonstrated that the Ciona GnRH-1 gene encoding Ciona GnRHs (t-GnRH-3, -5 and -6) was co-expressed with the Ci-GnRH-X gene in neurons of the cerebral ganglion. Of particular interest is that Ci-GnRH-X exhibited moderate (10–50%) inhibitory activity against t-GnRHs at their cognate receptors. Ci-GnRH-X repressed the elevation of the intracellular calcium and cAMP production by t-GnRH-6 at Ci-GnRHR-1, and cAMP production by t-GnRH-3, and t-GnRH-5 via Ci-GnRHR-3 was also inhibited by Ci-GnRH-X. In contrast, no inhibitory effect of Ci-GnRH-X at Ci-GnRHR-2 was observed. The localization and biochemical assays revealed that Ci-GnRH-X acts as an endogenous antagonist for the Ciona GnRHergic system. This is the first molecular and functional characterization of an endogenous inhibitor of GnRHs in an animal species.