1. Functional overexpression of vomeronasal receptors using a Herpes simplex virus type 1 (HSV-1)-derived amplicon
- Author
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Frank Zufall, Maria Teresa Alonso, Benjamin Stein, Pablo Chamero, Trese Leinders-Zufall, Department of Physiology, and Center for Integrative Physiology and Molecular Medicine, Saarland University [Saarbrücken], Instituto de Biología y Genética Molecular (IBGM), Universitad de Valladolid, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), 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, Center for Integrative Physiology and Molecular Medicine, Instituto de Biologia y Genetica Molecular, Universidad de Valladolid y Consejo Superior de Investigaciones Cientificas, Deutsche Forschungsgemeinschaft (SFB 894 A16 to TLZ and A17 to FZ, CH920/2-1), Volkswagen Foundation (TLZ), Saarland University, German Research Foundation, Volkswagen Foundation, Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), ProdInra, Archive Ouverte, Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), and European Chemoreception Research Organization (ECRO). INT.
- Subjects
0301 basic medicine ,Male ,Sensory Receptors ,Vomeronasal organ ,Epidemiology ,viruses ,Social Sciences ,lcsh:Medicine ,Herpesvirus 1, Human ,Disease Vectors ,souris ,Pathology and Laboratory Medicine ,Ligands ,Receptors, Odorant ,comportement animal ,social behaviour ,0302 clinical medicine ,Animal Cells ,Medicine and Health Sciences ,Psychology ,Receptor ,lcsh:Science ,Cells, Cultured ,Calcium signaling ,Neurons ,Multidisciplinary ,Organic Compounds ,Amplicon ,Chemoreceptor Cells ,3. Good health ,Cell biology ,Chemistry ,Medical Microbiology ,Viral Pathogens ,Second messenger system ,Viruses ,Physical Sciences ,Herpes Simplex Virus-1 ,Cell lines ,Sensory Perception ,Steroids ,Cellular Types ,Pathogens ,Viral Vectors ,Vomeronasal Organ ,Biological cultures ,Research Article ,Signal Transduction ,Autre (Sciences du Vivant) ,Herpesviruses ,[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT] ,mice ,Jacobson's organ ,virus de l'herpes ,vomeronasal ,Biology ,Microbiology ,Olfactory Receptor Neurons ,03 medical and health sciences ,Vomeronasal receptor ,GTP-Binding Proteins ,Virology ,Animals ,Humans ,organe de jacobson ,Calcium Signaling ,Molecular Biology Techniques ,phéromone ,Microbial Pathogens ,Molecular Biology ,comportement social ,signal chimique ,[SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT] ,HEK 293 cells ,Organic Chemistry ,Neuropeptides ,lcsh:R ,Organisms ,Chemical Compounds ,Biology and Life Sciences ,Afferent Neurons ,Cell Biology ,Molecular biology ,Herpes Simplex Virus ,Research and analysis methods ,Mice, Inbred C57BL ,030104 developmental biology ,HEK293 Cells ,Cellular Neuroscience ,lcsh:Q ,Heterologous expression ,DNA viruses ,030217 neurology & neurosurgery ,Viral Transmission and Infection ,Neuroscience ,Cloning - Abstract
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 Ca 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 Ca 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., This work was supported by Deutsche Forschungsgemeinschaft grants CH 920/2-1 (PC) and Sonderforschungsbereich 894 projects A16 (TL-Z) and A17 (FZ), a University of Saarland HOMFORexzellent grant (PC), the Volkswagen Foundation (TL-Z).
- Published
- 2016
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