Shark rectal gland vasoactive intestinal peptide receptor: cloning, functional expression, and regulation of CFTR chloride channels

Department of Medicine, Yale University School of Medicine, New Haven, Connecticut; and the Mount Desert Island Biological Laboratory, Salisbury Cove, Maine Submitted 29 January 2006 ; accepted in final form 8 May 2006 Vasoactive intestinal peptide (VIP) is a secretagogue that mediates chloride secretion in intestinal epithelia. We determined the relative potency of VIP and related peptides in the rectal gland of the elasmobranch dogfish shark and cloned and expressed the VIP receptor (sVIP-R) from this species. In the perfused rectal gland, VIP (5 nM) stimulated chloride secretion from 250 ± 66 to 2,604 ± 286 µeq·h –1 ·g –1 ; the relative potency of peptide agonists was VIP > PHI = GHRH > PACAP > secretin, where PHI is peptide histidine isoleucine amide, GHRH is growth hormone-releasing hormone, and PACAP is pituitary adenylate cylase activating peptide. The cloned sVIP-R from shark rectal gland (SRG) is only 61% identical to the human VIP-R1. It maintains a long, extracellular NH 2 terminus with seven cysteine residues, and has three N -glycosylation sites and eight other residues implicated in VIP binding. Two amino acids considered important for peptide binding in mammals are not present in the shark orthologue. When sVIP-R and the CFTR chloride channel were coexpressed in Xenopus oocytes, VIP increased chloride conductance from 11.3 ± 2 to 127 ± 34 µS. The agonist affinity for activating chloride conductance by the cloned receptor was VIP > GHRH = PHI > PACAP > secretin, a profile mirroring that in the perfused gland. The receptor differs from previously cloned VIP-Rs in having a low affinity for PACAP. Expression of both sVIP-R and CFTR mRNA was detected by quantitative PCR in shark rectal gland, intestine, and brain. These studies characterize a unique G protein-coupled receptor from the shark rectal gland that is the oldest cloned VIP-R. Squalus acanthias ; molecular cloning; Xenopus oocytes Address for reprint requests and other correspondence: J. N. Forrest, Jr., Dept. of Internal Medicine, Yale Univ. School of Medicine, New Haven, Connecticut 06510 (e-mail: john.forrest{at}yale.edu )