title: Mechanisms of protease-activated receptor 2-evoked hyperexcitability of nociceptive neurons innervating the mouse colon.
abstract: Agonists of protease-activated receptor 2 (PAR(2)) evoke hyperexcitability of dorsal root ganglia (DRG) neurons by unknown mechanisms. We examined the cellular mechanisms underlying PAR(2)-evoked hyperexcitability of mouse colonic DRG neurons to determine their potential role in pain syndromes such as visceral hyperalgesia. Colonic DRG neurons were identified by injecting Fast Blue and DiI retrograde tracers into the mouse colon. Using immunofluorescence, we found that DiI-labelled neurons contained PAR(2) immunoreactivity, confirming the presence of receptors on colonic neurons. Whole-cell current-clamp recordings of acutely dissociated neurons demonstrated that PAR(2) activation with a brief application (3 min) of PAR(2) agonists, SLIGRL-NH(2) and trypsin, evoked sustained depolarizations (up to 60 min) which were associated with increased input resistance and a marked reduction in rheobase (50% at 30 min). In voltage clamp, SLIGRL-NH(2) markedly suppressed delayed rectifier I(K) currents (55% at 10 min), but had no effect on the transient I(A) current or TTX-resistant Na(+) currents. In whole-cell current-clamp recordings, the sustained excitability evoked by PAR(2) activation was blocked by the PKC inhibitor, calphostin, and the ERK(1/2) inhibitor PD98059. Studies of ERK(1/2) phosphorylation using confocal microscopy demonstrated that SLIGRL-NH(2) increased levels of immunoreactive pERK(1/2) in DRG neurons, particularly in proximity to the plasma membrane. Thus, activation of PAR(2) receptors on colonic nociceptive neurons causes sustained hyperexcitability that is related, at least in part, to suppression of delayed rectifier I(K) currents. Both PKC and ERK(1/2) mediate the PAR(2)-induced hyperexcitability. These studies describe a novel mechanism of sensitization of colonic nociceptive neurons that may be implicated in conditions of visceral hyperalgesia such as irritable bowel syndrome.
authors.affiliation: Gastrointestinal Diseases Research Unit, Queen's University, Kingston, Ontario, Canada.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Kayssi
authors.forename: Silvia
authors.initials: S
authors.lastname: Amadesi
authors.forename: Francisco
authors.initials: F
authors.lastname: Bautista
authors.forename: Nigel W
authors.initials: NW
authors.lastname: Bunnett
authors.forename: Stephen
authors.initials: S
authors.lastname: Vanner
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003106:Colon / Q000294:innervation*; D004351:Drug Resistance; D004553:Electric Conductivity; D004789:Enzyme Activation / Q000502:physiology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009474:Neurons / Q000378:metabolism / Q000502:physiology; D009619:Nociceptors / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010766:Phosphorylation / Q000187:drug effects; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism; D044464:Receptor, PAR-2 / Q000819:agonists / Q000502:physiology*; D015222:Sodium Channels / Q000187:drug effects / Q000502:physiology; D013779:Tetrodotoxin / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
chemical_list: D009842:Oligopeptides; D015221:Potassium Channels; D044464:Receptor, PAR-2; D015222:Sodium Channels; C112356:seryl-leucyl-isoleucyl-glycyl--arginyl-leucinamide; D013779:Tetrodotoxin; D011493:Protein Kinase C; D048049:Extracellular Signal-Regulated MAP Kinases; D014357:Trypsin