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¹ Laboratoire de Neuroimmunologie des Annélides, UMR 8017 CNRS, SN3, Université des Sciences et Technologies de Lille, 59655 Villeneuve d’Ascq Cedex, France
² U469 INSERM, CCIPE, 141 rue de la Cardonille, 34094 Montpellier Cedex 5, France
³ Laboratoire de Neuroendocrinologie du Développement, UPRES-EA 2701, SN4, Université des Sciences et Technologies de Lille, 59655 Villeneuve d’Ascq Cedex, France

(Requests for offprints should be addressed to C Breton; Email: Christophe.Breton{at}univ-lille1.fr)

A Levoye is now at Institut Cochin, UMR 8104 CNRS, U567 INSERM, Université Paris V, Département de Biologie Cellulaire, 22 rue Méchain, 75014 Paris, France

^* (A Levoye and B Mouillac contributed equally to this work)

In annelids, it has been established that arginine-vasopressin (AVP)/oxytocin (OT) superfamily peptides are involved in the maintenance of water and electrolyte homeostasis as well as reproduction. At present, there is little information on their receptors. In this study, we report the characterization of a 1.7 kb cDNA for an AVP-related receptor from the leech Theromyzon tessulatum. The open reading frame encodes a 435-aminoacid transmembrane protein that displays seven segments of hydrophobic amino acids, typical of G-protein-coupled receptors. The overall predicted protein exhibits about 30% amino-acid identities to other invertebrate, as well as vertebrate, AVP/OT receptor family members, and displays conserved characteristic features belonging to the AVP/OT receptor superfamily. RT-PCR expression experiments showed that mRNA is expressed in the genital tract, the ovary and the brain. The receptor expression is stage specific, showing a weak expression after the two first blood meals, increasing dramatically after the last blood meal during the period of sexual maturation and disappearing after egg laying. Thus, the leech AVP-related receptor may mediate reproductive functions. When expressed in COS-7 cells, the receptor binds ligands with the following rank order of potency: AVP= Arg-vasotocin >Arg-conopressin >mesotocin = OT = Lys-conopressin=isotocin>annetocin. This shows an AVP-like pharmacological profile. The transfected receptor mediates AVP-induced accumulation of inositol phosphates, indicating that the leech AVP-related receptor is functional. This study describes the characterization of a novel AVP/OT superfamily receptor in annelids, which are considered the most distant group of coelomate metazoans possessing a functional AVP/OT-related endocrine system.

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