title: Transient receptor potential vanilloid 4 mediates protease activated receptor 2-induced sensitization of colonic afferent nerves and visceral hyperalgesia.
abstract: Protease-activated receptor (PAR(2)) is expressed by nociceptive neurons and activated during inflammation by proteases from mast cells, the intestinal lumen, and the circulation. Agonists of PAR(2) cause hyperexcitability of intestinal sensory neurons and hyperalgesia to distensive stimuli by unknown mechanisms. We evaluated the role of the transient receptor potential vanilloid 4 (TRPV4) in PAR(2)-induced mechanical hyperalgesia of the mouse colon. Colonic sensory neurons, identified by retrograde tracing, expressed immunoreactive TRPV4, PAR(2), and calcitonin gene-related peptide and are thus implicated in nociception. To assess nociception, visceromotor responses (VMR) to colorectal distension (CRD) were measured by electromyography of abdominal muscles. In TRPV4(+/+) mice, intraluminal PAR(2) activating peptide (PAR(2)-AP) exacerbated VMR to graded CRD from 6-24 h, indicative of mechanical hyperalgesia. PAR(2)-induced hyperalgesia was not observed in TRPV4(-/-) mice. PAR(2)-AP evoked discharge of action potentials from colonic afferent neurons in TRPV4(+/+) mice, but not from TRPV4(-/-) mice. The TRPV4 agonists 5',6'-epoxyeicosatrienoic acid and 4alpha-phorbol 12,13-didecanoate stimulated discharge of action potentials in colonic afferent fibers and enhanced current responses recorded from retrogradely labeled colonic dorsal root ganglia neurons, confirming expression of functional TRPV4. PAR(2)-AP enhanced these responses, indicating sensitization of TRPV4. Thus TRPV4 is expressed by primary spinal afferent neurons innervating the colon. Activation of PAR(2) increases currents in these neurons, evokes discharge of action potentials from colonic afferent fibers, and induces mechanical hyperalgesia. These responses require the presence of functional TRPV4. Therefore, TRPV4 is required for PAR(2)-induced mechanical hyperalgesia and excitation of colonic afferent neurons.
authors.affiliation: Department of Pediatrics, University of California San Francisco, San Francisco, CA 94143-0660, USA.
authors.forename: Walter E B
authors.initials: WE
authors.lastname: Sipe
authors.forename: Stuart M
authors.initials: SM
authors.lastname: Brierley
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Martin
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Phillis
authors.forename: Francisco Bautista
authors.initials: FB
authors.lastname: Cruz
authors.forename: Eileen F
authors.initials: EF
authors.lastname: Grady
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Liedtke
authors.forename: David M
authors.initials: DM
authors.lastname: Cohen
authors.forename: Stephen
authors.initials: S
authors.lastname: Vanner
authors.forename: L Ashley
authors.initials: LA
authors.lastname: Blackshaw
authors.forename: Nigel W
authors.initials: NW
authors.lastname: Bunnett
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis; D003106:Colon / Q000294:innervation / Q000503:physiopathology; D004576:Electromyography; D005260:Female; D005727:Ganglia, Spinal / Q000737:chemistry / Q000187:drug effects / Q000502:physiology; D006930:Hyperalgesia / Q000503:physiopathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology*; D009619:Nociceptors / Q000737:chemistry / Q000187:drug effects / Q000502:physiology; D010703:Phorbol Esters / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D044464:Receptor, PAR-2 / Q000819:agonists / Q000032:analysis / Q000502:physiology*; D012430:Ruthenium Red / Q000494:pharmacology; D012704:Serous Membrane / Q000294:innervation; D050052:TRPC Cation Channels / Q000819:agonists / Q000037:antagonists & inhibitors / Q000502:physiology*; D014781:Viscera / Q000294:innervation / Q000503:physiopathology
chemical_list: D010703:Phorbol Esters; D044464:Receptor, PAR-2; D050052:TRPC Cation Channels; C489571:TRPC4 ion channel; D012430:Ruthenium Red; C008876:phorbol-12,13-didecanoate; C040776:5,6-epoxy-8,11,14-eicosatrienoic acid; D015126:8,11,14-Eicosatrienoic Acid; D015740:Calcitonin Gene-Related Peptide