Journal Article

Functional overexpression of vomeronasal receptors using a Herpes simplex virus type 1 (HSV-1)-derived amplicon

Stein, Benjamin, Instituto de Biología y Genética Molecular (IBGM) ; Universitad de Valladolid, Zufall, Frank, Alonso, Maria Teresa, Leinders-Zufall, Trese, Physiologie de la reproduction et des comportements [Nouzilly] (PRC) ; Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Department of Physiology, and Center for Integrative Physiology and Molecular Medicine ; Universität des Saarlandes [Saarbrücken] = Saarland University [Saarbrücken], Chamero-Benito, Pablo, Deutsche Forschungsgemeinschaft grants CH 920/2-1 (PC); Sonderforschungsbereich 894 projects A16 (TL- Z) and A17 (FZ); University of Saarland HOMFORexzellent grant (PC); the Volkswagen Foundation (TL-Z). TL-Z is a Lichtenberg Professor of the Volkswagen Foundation

2016

Overview

In mice, social behaviors such as mating and aggression are mediated by pheromones and related chemosignals. The vomeronasal organ (VNO) detects olfactory information from other individuals by sensory neurons tuned to respond to specific chemical cues. Receptors expressed by vomeronasal neurons are implicated in selective detection of these cues. Nearly 400 receptor genes have been identified in the mouse VNO, but the tuning properties of individual receptors remain poorly understood, in part due to the lack of a robust heterologous expression system. Here we develop a herpes virus-based amplicon delivery system to overexpress three types of vomeronasal receptor genes and to characterize cell responses to their proposed ligands. Through Ca2+ imaging in native VNO cells we show that virus-induced overexpression of V1rj2, V2r1b or Fpr3 caused a pronounced increase of responsivity to sulfated steroids, MHC-binding peptide or the synthetic hexapeptide W-peptide, respectively. Other related ligands were not recognized by infected individual neurons, indicating a high degree of selectivity by the overexpressed receptor. Removal of G-protein signaling eliminates Ca2+ responses, indicating that the endogenous second messenger system is essential for observing receptor activation. Our results provide a novel expression system for vomeronasal receptors that should be useful for understanding the molecular logic of VNO ligand detection. Functional expression of vomeronasal receptors and their deorphanization provides an essential requirement for deciphering the neural mechanisms controlling behavior.

• Publisher: CCSD,Public Library of Science,Public Library of Science (PLoS)
• Subject: Animal care / Animals / Biology and Life Sciences / Calcium imaging / Calcium ions / Calcium Signaling / Calcium signalling / Cells, Cultured / Chemical stimuli / Chemoreception / Chemoreceptor Cells - metabolism / Development and progression / Gene expression / Genes / Genetic aspects / GTP-Binding Proteins - metabolism / HEK293 Cells / Herpes simplex / Herpes simplex virus 1 / Herpes viruses / Herpesvirus 1, Human - genetics / Herpesvirus infections / Humans / Life Sciences / Ligands / Major histocompatibility complex / Male / Medicine / Medicine and Health Sciences / Mice / Mice, Inbred C57BL / Neurons / Neuropeptides - metabolism / Odorant receptors / Other / Peptides / Pheromones / Physical Sciences / Physiological aspects / Physiology / Proteins / Receptor mechanisms / Receptors / Receptors, Odorant - genetics / Receptors, Odorant - metabolism / Research and analysis methods / Sensory neurons / Social aspects / Social behavior / Social Sciences / Steroid hormones / Steroids / Steroids - metabolism / Virology / Virus receptors / Viruses / Vomeronasal organ / Vomeronasal Organ - cytology / Vomeronasal Organ - metabolism / Vomeronasal system
• ISBN: 0003762911001