title: Modulation of Gq-protein-coupled inositol trisphosphate and Ca2+ signaling by the membrane potential.
abstract: Gq-protein-coupled receptors (GqPCRs) are widely distributed in the CNS and play fundamental roles in a variety of neuronal processes. Their activation results in phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis and Ca2+ release from intracellular stores via the phospholipase C (PLC)-inositol 1,4,5-trisphosphate (IP3) signaling pathway. Because early GqPCR signaling events occur at the plasma membrane of neurons, they might be influenced by changes in membrane potential. In this study, we use combined patch-clamp and imaging methods to investigate whether membrane potential changes can modulate GqPCR signaling in neurons. Our results demonstrate that GqPCR signaling in the human neuronal cell line SH-SY5Y and in rat cerebellar granule neurons is directly sensitive to changes in membrane potential, even in the absence of extracellular Ca2+. Depolarization has a bidirectional effect on GqPCR signaling, potentiating thapsigargin-sensitive Ca2+ responses to muscarinic receptor activation but attenuating those mediated by bradykinin receptors. The depolarization-evoked potentiation of the muscarinic signaling is graded, bipolar, non-inactivating, and with no apparent upper limit, ruling out traditional voltage-gated ion channels as the primary voltage sensors. Flash photolysis of caged IP3/GPIP2 (glycerophosphoryl-myo-inositol 4,5-bisphosphate) places the voltage sensor before the level of the Ca2+ store, and measurements using the fluorescent bioprobe eGFP-PH(PLCdelta) (enhanced green fluorescent protein-pleckstrin homology domain-PLCdelta) directly demonstrate that voltage affects muscarinic signaling at the level of the IP3 production pathway. The sensitivity of GqPCR IP3 signaling in neurons to voltage itself may represent a fundamental mechanism by which ionotropic signals can shape metabotropic receptor activity in neurons and influence processes such as synaptic plasticity in which the detection of coincident signals is crucial.
authors.affiliation: Department of Cell Physiology and Pharmacology, Medical Sciences Building, University of Leicester, Leicester LE1 9HN, United Kingdom. db84@le.ac.uk
authors.forename: Daniela
authors.initials: D
authors.lastname: Billups
authors.forename: Brian
authors.initials: B
authors.lastname: Billups
authors.forename: R A John
authors.initials: RA
authors.lastname: Challiss
authors.forename: Stefan R
authors.initials: SR
authors.lastname: Nahorski
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured / Q000502:physiology; D002531:Cerebellum / Q000166:cytology; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000502:physiology*; D007295:Inositol Phosphates / Q000528:radiation effects; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000196:embryology; D008564:Membrane Potentials / Q000502:physiology*; D008856:Microscopy, Fluorescence; D009447:Neuroblastoma / Q000473:pathology; D009473:Neuronal Plasticity; D009474:Neurons / Q000502:physiology*; D009543:Nifedipine / Q000494:pharmacology; D010095:Oxotremorine / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D054803:Phospholipase C delta; D010782:Photolysis; D051381:Rats; D043587:Receptor, Muscarinic M3 / Q000819:agonists / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology; D019284:Thapsigargin / Q000494:pharmacology; D014162:Transfection; D010738:Type C Phospholipases / Q000235:genetics / Q000378:metabolism
chemical_list: C087840:1-(glycerophosphoryl)-myo-inositol 4,5-diphosphate P(4(5))-1-(2-nitrophenyl)ethyl ester; D007295:Inositol Phosphates; D007527:Isoenzymes; D019269:Phosphatidylinositol 4,5-Diphosphate; D043587:Receptor, Muscarinic M3; D011993:Recombinant Fusion Proteins; D010095:Oxotremorine; D019284:Thapsigargin; D015544:Inositol 1,4,5-Trisphosphate; D010738:Type C Phospholipases; D054803:Phospholipase C delta; D009543:Nifedipine; D015056:1-Methyl-3-isobutylxanthine