Transient receptor potential vanilloid type 1 activation down-regulates voltage-gated calcium channels through calcium-dependent calcineurin in sensory neurons.
Calcium influx through voltage-activated Ca(2+) channels (VACCs) plays a critical role in neurotransmission. Capsaicin application inhibits VACCs and desensitizes nociceptors. In this study, we determined the signaling mechanisms of the inhibitory effect of capsaicin on VACCs in primary sensory neurons. Whole-cell voltage clamp recordings were performed in acutely isolated rat dorsal root ganglion neurons. Capsaicin caused a profound decrease in the Ca(2+) current (I(Ca)) density in capsaicin-sensitive, but not -insensitive, dorsal root ganglion neurons. At 1 mum, capsaicin suppressed about 60% of N-, P/Q-, L-, and R-type I(Ca) density. Pretreatment with iodoresiniferatoxin, a specific transient receptor potential vanilloid type 1 (TRPV1) antagonist, or intracellular application of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid blocked the inhibitory effect of capsaicin on I(ca). However, neither W-7, a calmodulin blocker, nor KN-93, a CaMKII inhibitor, attenuated the inhibitory effect of capsaicin on I(Ca). Furthermore, intracellular dialysis of deltamethrin or cyclosporin A, the specific calcineurin (protein phosphatase 2B) inhibitors, but not okadaic acid (a selective protein phosphatase 1/protein phosphatase 2A inhibitor), abolished the effect of capsaicin on I(Ca). Interestingly, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, deltamethrin, cyclosporin A, and okadaic acid each alone significantly increased the I(Ca) density and caused a depolarizing shift in the voltage dependence of activation. Immunofluorescence labeling revealed that capsaicin induced a rapid internalization of Ca(V)2.2 channels on the membrane. Thus, this study provides novel information that VACCs are tonically modulated by the intracellular Ca(2+) level and endogenous phosphatases in sensory neurons. Stimulation of TRPV1 by capsaicin down-regulates VACCs by dephosphorylation through Ca(2+)-dependent activation of calcineurin.
Wu
Zi-Zhen
ZZ
Department of Anesthesiology, Department of Neural and Behavioral Sciences, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
Chen
Shao-Rui
SR
Pan
Hui-Lin
HL
D000818:Animals; D019703:Calcineurin / Q000494:pharmacology*; D015220:Calcium Channels / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology*; D007473:Ion Channels / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009475:Neurons, Afferent / Q000187:drug effects / Q000378:metabolism*; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2; D051381:Rats; D017207:Rats, Sprague-Dawley; D050916:TRPV Cation Channels
D015220:Calcium Channels; D007473:Ion Channels; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat; D019703:Calcineurin; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2