title: TRPV4 forms a novel Ca2+ signaling complex with ryanodine receptors and BKCa channels.
abstract: Vasodilatory factors produced by the endothelium are critical for the maintenance of normal blood pressure and flow. We hypothesized that endothelial signals are transduced to underlying vascular smooth muscle by vanilloid transient receptor potential (TRPV) channels. TRPV4 message was detected in RNA from cerebral artery smooth muscle cells. In patch-clamp experiments using freshly isolated cerebral myocytes, outwardly rectifying whole-cell currents with properties consistent with those of expressed TRPV4 channels were evoked by the TRPV4 agonist 4alpha-phorbol 12,13-didecanoate (4alpha-PDD) (5 micromol/L) and the endothelium-derived arachidonic acid metabolite 11,12 epoxyeicosatrienoic acid (11,12 EET) (300 nmol/L). Using high-speed laser-scanning confocal microscopy, we found that 11,12 EET increased the frequency of unitary Ca2+ release events (Ca2+ sparks) via ryanodine receptors located on the sarcoplasmic reticulum of cerebral artery smooth muscle cells. EET-induced Ca2+ sparks activated nearby sarcolemmal large-conductance Ca2+-activated K+ (BKCa) channels, measured as an increase in the frequency of transient K+ currents (referred to as "spontaneous transient outward currents" [STOCs]). 11,12 EET-induced increases in Ca2+ spark and STOC frequency were inhibited by lowering external Ca2+ from 2 mmol/L to 10 micromol/L but not by voltage-dependent Ca2+ channel inhibitors, suggesting that these responses require extracellular Ca2+ influx via channels other than voltage-dependent Ca2+ channels. Antisense-mediated suppression of TRPV4 expression in intact cerebral arteries prevented 11,12 EET-induced smooth muscle hyperpolarization and vasodilation. Thus, we conclude that TRPV4 forms a novel Ca2+ signaling complex with ryanodine receptors and BKCa channels that elicits smooth muscle hyperpolarization and arterial dilation via Ca2+-induced Ca2+ release in response to an endothelial-derived factor.
authors.affiliation: Department of Pharmacology, University of Vermont College of Medicine, Burlington, VT 05405, USA. Scott.Earley@uvm.edu
authors.forename: Scott
authors.initials: S
authors.lastname: Earley
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Heppner
authors.forename: Mark T
authors.initials: MT
authors.lastname: Nelson
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Brayden
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D002536:Cerebral Arteries / Q000737:chemistry; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000502:physiology*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000737:chemistry / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019837:Ryanodine Receptor Calcium Release Channel / Q000502:physiology*; D050916:TRPV Cation Channels / Q000032:analysis / Q000502:physiology*; D014664:Vasodilation / Q000187:drug effects
chemical_list: D051036:Large-Conductance Calcium-Activated Potassium Channels; D019837:Ryanodine Receptor Calcium Release Channel; D050916:TRPV Cation Channels; C486577:Trpv4 protein, rat; C046783:11,12-epoxy-5,8,14-eicosatrienoic acid; D015126:8,11,14-Eicosatrienoic Acid; D002118:Calcium