title: The cannabinoid WIN 55,212-2 inhibits transient receptor potential vanilloid 1 (TRPV1) and evokes peripheral antihyperalgesia via calcineurin.
abstract: Cannabinoids can evoke antihyperalgesia and antinociception at a peripheral site of action. However, the signaling pathways mediating these effects are not clearly understood. We tested the hypothesis that certain cannabinoids directly inhibit peripheral capsaicin-sensitive nociceptive neurons by dephosphorylating and desensitizing transient receptor potential vanilloid 1 (TRPV1) via a calcium calcineurin-dependent mechanism. Application of the cannabinoid WIN 55,212-2 (WIN) to cultured trigeminal (TG) neurons or isolated skin biopsies rapidly and significantly inhibited capsaicin-activated inward currents and neuropeptide exocytosis by a mechanism requiring the presence of extracellular calcium. The inhibitory effect did not involve activation of G protein-coupled cannabinoid receptors, because neither pertussis toxin nor GDPbetaS treatments altered the WIN effect. However, application of WIN-activated calcineurin, as measured by nuclear translocation of the nuclear factor of activated T cells (NFAT)c4 transcription factor, dephosphorylated TRPV1. The WIN-induced desensitization of TRPV1 was mediated by calcineurin, because the application of structurally distinct calcineurin antagonists (calcineurin autoinhibitory peptide and cyclosporine/cyclophilin complex) abolished WIN-induced inhibition of capsaicin-evoked inward currents and neuropeptide exocytosis. This mechanism also contributed to peripheral antinociceptive/antihyperalgesic effects of WIN because pretreatment with the calcineurin antagonist calcineurin autoinhibitory peptide (CAIP) significantly reduced peripherally mediated WIN effects in two behavioral models. Collectively, these data demonstrate that cannabinoids such as WIN directly inhibit TRPV1 functional activities via a calcineurin pathway that represents a mechanism of cannabinoid actions at peripheral sites.
authors.affiliation: Department of Pharmacology, University of Texas Health Science Center, San Antonio, TX 78229, USA.
authors.forename: Amol M
authors.initials: AM
authors.lastname: Patwardhan
authors.forename: Nathaniel A
authors.initials: NA
authors.lastname: Jeske
authors.forename: Theodore J
authors.initials: TJ
authors.lastname: Price
authors.forename: Nikita
authors.initials: N
authors.lastname: Gamper
authors.forename: Armen N
authors.initials: AN
authors.lastname: Akopian
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Hargreaves
mesh_terms: D000818:Animals; D048588:Benzoxazines; D019703:Calcineurin / Q000378:metabolism / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D006930:Hyperalgesia / Q000188:drug therapy*; D008297:Male; D009025:Morpholines / Q000494:pharmacology*; D050778:NFATC Transcription Factors / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology*; D009474:Neurons / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012867:Skin / Q000187:drug effects; D050916:TRPV Cation Channels / Q000037:antagonists & inhibitors*
chemical_list: D048588:Benzoxazines; D002121:Calcium Channel Blockers; D009025:Morpholines; D050778:NFATC Transcription Factors; D009281:Naphthalenes; D010455:Peptides; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat; C070417:(3R)-((2,3-dihydro-5-methyl-3-((4-morpholinyl)methyl)pyrrolo-(1,2,3-de)-1,4-benzoxazin-6-yl)(1-naphthalenyl))methanone; D019703:Calcineurin; D002118:Calcium