title: Role of anoctamin-1 and bestrophin-1 in spinal nerve ligation-induced neuropathic pain in rats.
abstract: BACKGROUND
  Calcium-activated chloride channels (CaCCs) activation induces membrane depolarization by increasing chloride efflux in primary sensory neurons that can facilitate action potential generation. Previous studies suggest that CaCCs family members bestrophin-1 and anoctamin-1 are involved in inflammatory pain. However, their role in neuropathic pain is unclear. In this investigation we assessed the involvement of these CaCCs family members in rats subjected to the L5/L6 spinal nerve ligation. In addition, anoctamin-1 and bestrophin-1 mRNA and protein expression in dorsal root ganglion (DRG) and spinal cord was also determined in the presence and absence of selective inhibitors.
  
  
  RESULTS
  L5/L6 spinal nerve ligation induced mechanical tactile allodynia. Intrathecal administration of non-selective CaCCs inhibitors (NPPB, 9-AC and NFA) dose-dependently reduced tactile allodynia. Intrathecal administration of selective CaCCs inhibitors (T16Ainh-A01 and CaCCinh-A01) also dose-dependently diminished tactile allodynia and thermal hyperalgesia. Anoctamin-1 and bestrophin-1 mRNA and protein were expressed in the dorsal spinal cord and DRG of naïve, sham and neuropathic rats. L5/L6 spinal nerve ligation rose mRNA and protein expression of anoctamin-1, but not bestrophin-1, in the dorsal spinal cord and DRG from day 1 to day 14 after nerve ligation. In addition, repeated administration of CaCCs inhibitors (T16Ainh-A01, CaCCinh-A01 or NFA) or anti-anoctamin-1 antibody prevented spinal nerve ligation-induced rises in anoctamin-1 mRNA and protein expression. Following spinal nerve ligation, the compound action potential generation of putative C fibers increased while selective CaCCs inhibitors (T16Ainh-A01 and CaCCinh-A01) attenuated such increase.
  
  
  CONCLUSIONS
  There is functional anoctamin-1 and bestrophin-1 expression in rats at sites related to nociceptive processing. Blockade of these CaCCs suppresses compound action potential generation in putative C fibers and lessens established tactile allodynia. As CaCCs activity contributes to neuropathic pain maintenance, selective inhibition of their activity may function as a tool to generate analgesia in nerve injury pain states.
authors.affiliation: Neurobiology of Pain Laboratory, Departamento de Farmacobiología, Centro de Investigación y de Estudios Avanzados (Cinvestav), Sede Sur, Calzada de los Tenorios 235, Colonia Granjas Coapa, 14330, México, D.F., México. jorgepinedafarias@yahoo.com.mx.
authors.forename: Jorge Baruch
authors.initials: JB
authors.lastname: Pineda-Farias
authors.affiliation: Neurobiology of Pain Laboratory, Departamento de Farmacobiología, Centro de Investigación y de Estudios Avanzados (Cinvestav), Sede Sur, Calzada de los Tenorios 235, Colonia Granjas Coapa, 14330, México, D.F., México. baip.neurofarm@gmail.com.
authors.forename: Paulino
authors.initials: P
authors.lastname: Barragán-Iglesias
authors.affiliation: Departamento de Fisiología, Biofísica y Neurociencias, Centro de Investigación y de Estudios Avanzados (Cinvestav), Zacatenco, México, D.F., México. loezayuc@fisio.cinvestav.mx.
authors.forename: Emanuel
authors.initials: E
authors.lastname: Loeza-Alcocer
authors.affiliation: Laboratorio de Mecanismos de Dolor, División Académica de Ciencias de la Salud, Universidad Juárez Autónoma de Tabasco, Villahermosa, Tabasco, México. jetorreslopez@hotmail.com.|Hospital Regional de Alta Especialidad "Dr. Juan Graham Casasús", Villahermosa, Tabasco, México. jetorreslopez@hotmail.com.
authors.forename: Jorge E
authors.initials: JE
authors.lastname: Torres-López
authors.affiliation: Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina, Instituto Politécnico Nacional, México, D.F., México. hector.isaac@gmail.com.
authors.forename: Héctor Isaac
authors.initials: HI
authors.lastname: Rocha-González
authors.affiliation: Departamento de Neuroquímica, Instituto Nacional de Neurología y Neurocirugía "Manuel Velasco Suárez", México, D.F., México. fseverian@yahoo.com.
authors.forename: Francisca
authors.initials: F
authors.lastname: Pérez-Severiano
authors.affiliation: Departamento de Fisiología, Biofísica y Neurociencias, Centro de Investigación y de Estudios Avanzados (Cinvestav), Zacatenco, México, D.F., México. rdelgado@fisio.cinvestav.mx.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Delgado-Lezama
authors.affiliation: Neurobiology of Pain Laboratory, Departamento de Farmacobiología, Centro de Investigación y de Estudios Avanzados (Cinvestav), Sede Sur, Calzada de los Tenorios 235, Colonia Granjas Coapa, 14330, México, D.F., México. vgranados@prodigy.net.mx.
authors.forename: Vinicio
authors.initials: V
authors.lastname: Granados-Soto
mesh_terms: D000818:Animals; D000075369:Anoctamin-1; D000075002:Bestrophins; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006930:Hyperalgesia / Q000150:complications / Q000473:pathology / Q000503:physiopathology; D007278:Injections, Spinal; D008026:Ligation; D009043:Motor Activity; D009431:Neural Conduction; D009437:Neuralgia / Q000150:complications / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017208:Rats, Wistar; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D013127:Spinal Nerves / Q000293:injuries / Q000473:pathology* / Q000503:physiopathology
chemical_list: C552942:ANO1 protein, rat; D000075369:Anoctamin-1; C508904:Best1 protein, rat; D000075002:Bestrophins; D018118:Chloride Channels; D012333:RNA, Messenger