Modality-specific mechanisms of protein kinase C-induced hypersensitivity of TRPV1: S800 is a polymodal sensitization site.
TRPV1 is a nociceptive ion channel activated by polymodal stimuli such as capsaicin, proton, and noxious heat. Multiple inflammatory mediators activate protein kinases, especially protein kinase C (PKC), which phosphorylates TRPV1. Emerging evidence suggests that phosphorylation of TRPV1 constitutes specific signals underpinning pathological nociception. Although the mechanisms of hypersensitivity of TRPV1 to capsaicin are well studied, the phosphorylation residues that contribute to hypersensitivity to heat or acid have not been identified. In this study, we investigated modality-specific mechanisms of PKC-induced hypersensitivity using mutagenic ablation of PKC-associated phosphorylation sites in TRPV1. In heterologous systems, TRPV1 S502 and S800, but not T704, are known to be involved in hypersensitivity to capsaicin after the application of phorbol myristate acetate (PMA), a PKC agonist. Unlike capsaicin, PMA-induced hypersensitivity to heat was attenuated in TRPV1 mutants T704A and S800A, but not in S502A. In contrast, PMA-induced hypersensitivity to acid was attenuated only in S800A. To examine the roles of these phosphorylation sites in more physiologically relevant conditions, TRPV1 and mutants were tested in sensory neurons from TRPV1-null mice. In sensory neurons expressing mutated TRPV1, we found that alanine mutation of S800 commonly attenuates PMA-induced hypersensitivity to capsaicin, heat, and acid. Moreover, bradykinin-induced hypersensitivity to capsaicin was largely attenuated by the S800A mutation. These results suggest that mechanisms of PKC-induced hypersensitivity of TRPV1 are modality specific and that S800 is a polymodal sensitization site integrating multiple inflammatory signals in nociceptors. Our data provide a rationale for a novel approach targeting TRPV1 S800 for antihyperalgesia.
Department of Neural and Pain Sciences, the University of Maryland, Program in Neuroscience, School of Dentistry, Baltimore, MD, USA.
Sen
S
Wang
John
J
Joseph
Jin Y
JY
Ro
Man-Kyo
MK
Chung
D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002211:Capsaicin / Q000494:pharmacology; D002461:Cell Line, Transformed; D057809:HEK293 Cells; D006358:Hot Temperature / Q000009:adverse effects; D006801:Humans; D006967:Hypersensitivity / Q000378:metabolism* / Q000503:physiopathology; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D059225:Nociception / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D010766:Phosphorylation / Q000187:drug effects*; D011493:Protein Kinase C / Q000378:metabolism*; D011984:Sensory Receptor Cells / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002211:Capsaicin; D001920:Bradykinin