Functional characterisation of neuropeptides that act as ligands for both calcitonin-type and pigment-dispersing factor-type receptors in a deuterostome

The calcitonin (CT) family of related peptides exerts diverse physiological effects in mammals via two G-protein coupled receptors: CTR and the CTR-like receptor CLR. Phylogenetic analysis of CT-type signaling has revealed the presence of CT-type peptides and CTR/CLR-type proteins in both deuterostome and protostome invertebrates. Furthermore, experimental studies have demonstrated that in the protostomeDrosophila melanogasterthe CT-like peptide DH₃₁can act as a ligand for a CTR/CLR-type receptor and a pigment-dispersing factor (PDF) receptor. Here we investigated the signaling mechanisms and functions of CT-type neuropeptides in a deuterostome invertebrate, the sea cucumberApostichopus japonicus(phylum Echinodermata). InA. japonicus,a single gene encodes two CT-type peptides (AjCT1 and AjCT2) and both peptides act as ligands for a CTR/CLR-type receptor (AjCTR) and two PDF-type receptors (AjPDFR1, AjPDFR2), but with differential activation of downstream cAMP/PKA, Gαq/Ca²⁺/PKC and ERK1/2 signaling pathways. Transcriptic analysis of the AjCT1/AjCT2-encoding gene revealed expression in the central nervous system and a variety of organ systems, which was visualized immunohistochemically using an antiserum to a starfish CT-type peptide (ArCT).In vitropharmacological experiments demonstrated that AjCT1 and/or AjCT2 cause dose-dependent relaxation of longitudinal muscle and intestine preparations. Furthermore,in vivopharmacological experiments, combined with gain- and loss-of-function experiments, revealed a potential physiological role for AjCT2/AjPDFR2 signaling in promoting feeding and growth inA. japonicus. To our knowledge, this is the first study to obtain evidence that CT-type peptides can act as ligands for both CTR/CLR-type and PDF-type receptors in a deuterostome. Moreover, it provides the first evidence for appetite-stimulating and growth-promoting effects of CT-type neuropeptides in bilaterians. Given the economic importance ofA. japonicusas a foodstuff, the discovery of CT-type peptides as potential regulators of feeding and growth in this species may offer novel strategies for aquaculture applications.