GTP up-regulated persistent Na+ current and enhanced nociceptor excitability require NaV1.9.
Persistent tetrodotoxin-resistant (TTX-r) sodium currents up-regulated by intracellular GTP have been invoked as the site of action of peripheral inflammatory mediators that lower pain thresholds, and ascribed to the Na(V)1.9 sodium channel. Here we describe the properties of a global knock-out of Na(V)1.9 produced by replacing exons 4 and 5 in SCN11A with a neomycin resistance cassette, deleting the domain 1 voltage sensor and introducing a frameshift mutation. Recordings from small (< 25 microm apparent diameter) sensory neurones indicated that channel loss eliminates a TTX-r persistent current. Intracellular dialysis of GTP-gamma-S did not cause an up-regulation of persistent Na(+) current in Na(V)1.9-null neurones and the concomitant negative shift in voltage-threshold seen in wild-type and heterozygous neurones. Heterologous hNa(V)1.9 expression in Na(V)1.9 knock-out sensory neurones confirms that the human clone can restore the persistent Na(+) current. Taken together, these findings demonstrate that Na(V)1.9 underlies the G-protein pathway-regulated TTX-r persistent Na(+) current in small diameter sensory neurones that may drive spontaneous discharge in nociceptive nerve fibres during inflammation.
Ostman
Johan A R
JA
Centre for Neuroscience, Institute of Cell and Molecular Science, Queen Mary University of London, Newark Street, London E1 2AT, UK.
Nassar
Mohammed A
MA
Wood
John N
JN
Baker
Mark D
MD
D000200:Action Potentials / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D005091:Exons / Q000235:genetics; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D062558:NAV1.9 Voltage-Gated Sodium Channel; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009475:Neurons, Afferent / Q000166:cytology / Q000378:metabolism*; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism*; D009619:Nociceptors / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D062558:NAV1.9 Voltage-Gated Sodium Channel; D009479:Neuropeptides; C491885:Scn11a protein, mouse; D015222:Sodium Channels; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D012964:Sodium