Vanilloid receptor-1 (TRPV1)-dependent activation of inhibitory neurotransmission in spinal substantia gelatinosa neurons of mouse.
Inhibitory neurotransmission in spinal cord dorsal horn is mainly mediated by gamma-amino butyric acid (GABA) and glycine. By patch clamp recordings and correlative immunocytochemistry, we studied here the effect of 2 microM capsaicin-induced vanilloid receptor-1 (TRPV1) activation on IPSCs in spinal lamina II neurons from post-natal mice. Specificity was confirmed after pre-incubation with the competitive antagonist SB366791 (10 microM). After a single capsaicin pulse, an intense increase of spontaneous IPSC (sIPSC) frequency was observed in the presence of NBQX 10 microM (62/81 neurons; approximately 76%) or NBQX 10 microM + AP-5 20-100 microM (27/42 neurons; approximately 64%). Only a subpopulation (approximately 40%) of responsive neurons showed a significant amplitude increase. Seventy-two percent of the neurons displayed pure GABA(A) receptor-mediated sIPSCs, whereas the remaining ones showed mixed GABAergic/glycinergic events. After two consecutive capsaicin pulses, frequency rises were very similar, and both significantly higher than controls. When the second pulse was given in the presence of 4 microM L732,138, a selective antagonist of the substance P (SP) preferred receptor NK1, we observed a significant loss in frequency increase (63.90% with NBQX and 52.35% with NBQX + AP-5). TTX (1 microM) largely (approximately 81.5%) blocked the effect of capsaicin. These results show that TRPV1 activation on primary afferent fibers releases SP. The peptide then excites inhibitory neurons in laminae I, III and IV, leading to an increased release of GABA/glycine in lamina II via a parallel alternative pathway to glutamate.
Ferrini
Francesco
F
Department of Veterinary Morphophysiology, Via Leonardo da Vinci 44, 10095 Grugliasco, Italy.
Salio
Chiara
C
Vergnano
Angela Maria
AM
Merighi
Adalberto
A
D000813:Anilides / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D001640:Bicuculline / Q000494:pharmacology; D002211:Capsaicin / Q000494:pharmacology; D002934:Cinnamates / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D066298:In Vitro Techniques; D053444:Inhibitory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D051379:Mice; D009433:Neural Inhibition / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000379:methods; D013376:Substantia Gelatinosa / Q000166:cytology*; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*; D050916:TRPV Cation Channels / Q000037:antagonists & inhibitors / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014364:Tryptophan / Q000031:analogs & derivatives / Q000494:pharmacology
D000813:Anilides; D002934:Cinnamates; D018691:Excitatory Amino Acid Antagonists; C477659:N-(3-methoxyphenyl)-4-chlorocinnamanilide; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse; C086089:3,5-bis(trifluoromethyl)benzyl N-acetyltryptophan; D013779:Tetrodotoxin; D014364:Tryptophan; D002211:Capsaicin; D001640:Bicuculline