title: Intracellular alkalization causes pain sensation through activation of TRPA1 in mice.
abstract: Vertebrate cells require a very narrow pH range for survival. Cells accordingly possess sensory and defense mechanisms for situations where the pH deviates from the viable range. Although the monitoring of acidic pH by sensory neurons has been attributed to several ion channels, including transient receptor potential vanilloid 1 channel (TRPV1) and acid-sensing ion channels (ASICs), the mechanisms by which these cells detect alkaline pH are not well understood. Here, using Ca2+ imaging and patch-clamp recording, we showed that alkaline pH activated transient receptor potential cation channel, subfamily A, member 1 (TRPA1) and that activation of this ion channel was involved in nociception. In addition, intracellular alkalization activated TRPA1 at the whole-cell level, and single-channel openings were observed in the inside-out configuration, indicating that alkaline pH activated TRPA1 from the inside. Analyses of mutants suggested that the two N-terminal cysteine residues in TRPA1 were involved in activation by intracellular alkalization. Furthermore, intraplantar injection of ammonium chloride into the mouse hind paw caused pain-related behaviors that were not observed in TRPA1-deficient mice. These results suggest that alkaline pH causes pain sensation through activation of TRPA1 and may provide a molecular explanation for some of the human alkaline pH-related sensory disorders whose mechanisms are largely unknown.
authors.affiliation: Section of Cell Signaling, Okazaki Institute for Integrative Bioscience, National Institute of Natural Sciences, Okazaki, Japan.
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Fujita
authors.forename: Kunitoshi
authors.initials: K
authors.lastname: Uchida
authors.forename: Tomoko
authors.initials: T
authors.lastname: Moriyama
authors.forename: Asako
authors.initials: A
authors.lastname: Shima
authors.forename: Koji
authors.initials: K
authors.lastname: Shibasaki
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Inada
authors.forename: Takaaki
authors.initials: T
authors.lastname: Sokabe
authors.forename: Makoto
authors.initials: M
authors.lastname: Tominaga
mesh_terms: D000818:Animals; D001522:Behavior, Animal; D002118:Calcium / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008817:Mice, Mutant Strains; D017288:Pain Threshold; D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels / Q000235:genetics / Q000378:metabolism*
chemical_list: D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels; C475685:Trpa1 protein, mouse; D002118:Calcium