title: Activation of TRPA1 by membrane permeable local anesthetics.
abstract: BACKGROUND
  Low concentrations of local anesthetics (LAs) suppress cellular excitability by inhibiting voltage-gated Na⁺ channels. In contrast, LAs at high concentrations can be excitatory and neurotoxic. We recently demonstrated that LA-evoked activation of sensory neurons is mediated by the capsaicin receptor TRPV1, and, to a lesser extent by the irritant receptor TRPA1. LA-induced activation and sensitization of TRPV1 involves a domain that is similar, but not identical to the vanilloid-binding domain. Additionally, activation of TRPV1 by LAs involves PLC and PI(4,5)P₂-signalling. In the present study we aimed to characterize essential structural determinants for LA-evoked activation of TRPA1.
  
  
  RESULTS
  Recombinant rodent and human TRPA1 were expressed in HEK293t cells and investigated by means of whole-cell patch clamp recordings. The LA lidocaine activates TRPA1 in a concentration-dependent manner. The membrane impermeable lidocaine-derivative QX-314 is inactive when applied extracellularly. Lidocaine-activated TRPA1-currents are blocked by the TRPA1-antagonist HC-030031. Lidocaine is also an inhibitor of TRPA1, an effect that is more obvious in rodent than in human TRPA1. This species-specific difference is linked to the pore region (transmembrane domain 5 and 6) as described for activation of TRPA1 by menthol. Unlike menthol-sensitivity however, lidocaine-sensitivity is not similarly determined by serine- and threonine-residues within TM5. Instead, intracellular cysteine residues known to be covalently bound by reactive TRPA1-agonists seem to mediate activation of TRPA1 by LAs.
  
  
  CONCLUSIONS
  The structural determinants involved in activation of TRPA1 by LAs are disparate from those involved in activation by menthol or those involved in activation of TRPV1 by LAs.
authors.affiliation: Department of Anesthesiology, Friedrich-Alexander-University Erlangen-Nuremberg, Krankenhausstrasse 12, 91054 Erlangen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Leffler
authors.forename: Anja
authors.initials: A
authors.lastname: Lattrell
authors.forename: Sergej
authors.initials: S
authors.lastname: Kronewald
authors.forename: Florian
authors.initials: F
authors.lastname: Niedermirtl
authors.forename: Carla
authors.initials: C
authors.lastname: Nau
mesh_terms: D000779:Anesthetics, Local / Q000494:pharmacology*; D000818:Animals; D017487:Ankyrins / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D015640:Ion Channel Gating / Q000187:drug effects*; D008012:Lidocaine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D009149:Mustard Plant; D050505:Mutant Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010938:Plant Oils / Q000494:pharmacology; D051381:Rats; D000074025:TRPA1 Cation Channel; D050052:TRPC Cation Channels; D050053:TRPM Cation Channels / Q000378:metabolism; D050051:Transient Receptor Potential Channels / Q000378:metabolism*
chemical_list: D000779:Anesthetics, Local; D017487:Ankyrins; D015220:Calcium Channels; D050505:Mutant Proteins; D009419:Nerve Tissue Proteins; D010938:Plant Oils; D000074025:TRPA1 Cation Channel; C473350:TRPA1 protein, human; D050052:TRPC Cation Channels; D050053:TRPM Cation Channels; D050051:Transient Receptor Potential Channels; C475685:Trpa1 protein, mouse; C480976:Trpa1 protein, rat; C498947:Trpm8 protein, rat; C012647:QX-314; D008012:Lidocaine; D002118:Calcium; C027793:mustard oil