Association between Tetrodotoxin Resistant Channels and Lipid Rafts Regulates Sensory Neuron Excitability 英文参考文献.docVIP

AssociationbetweenTetrodotoxinResistantChannels andLipidRaftsRegulatesSensoryNeuronExcitability AlessandroPristera`1,MarkD.Baker2,KenjiOkuse1* 1Division of Cell and Molecular Biology, Faculty of Natural Sciences, Imperial College London, London, United Kingdom, 2Neuroscience and Trauma Centre, Blizard Institute,QueenMaryUniversityofLondon,BartsandTheLondonSchoolofMedicineandDentistry,London,UnitedKingdom Abstract Voltage-gatedsodiumchannels(VGSCs)playakeyroleintheinitiationandpropagationofactionpotentialsinneurons. NaV1.8isatetrodotoxin(TTX)resistantVGSCexpressedinnociceptors,peripheralsmall-diameterneuronsabletodetect noxious stimuli. NaV1.8 underlies the vast majority of sodium currents during action potentials. Many studies have highlighted a key role for NaV1.8 in inflammatory and chronic pain models. Lipid rafts are microdomains of the plasma membrane highly enriched in cholesterol and sphingolipids. Lipid rafts tune the spatial and temporal organisation of proteinsandlipidsontheplasmamembrane.Theyarethoughttoactasplatformsonthemembranewhereproteinsand lipids can be trafficked, compartmentalised and functionally clustered. In the present study we investigated NaV1.8 sub- cellular localisation and explored the idea that it is associated with lipid rafts in nociceptors. We found that NaV1.8 is distributed in clusters along the axons of DRG neurons in vitro and ex vivo. We also demonstrated, by biochemical and imaging studies, that NaV1.8 is associated with lipid rafts along the sciatic nerve ex vivo and in DRG neurons in vitro. Moreover,treatmentswithmethyl-b-cyclodextrin(MbCD)and7-ketocholesterol(7KC)ledtothedissociationbetweenrafts and NaV1.8. By calcium imaging wedemonstrated that the lack of association between rafts and NaV1.8 correlated with impaired neuronal excitability, highlighted by a reduction in the number of neurons able to conduct mechanically- and chemically-evokeddepolarisations.Thesefindingsrevealthesub-cellularlocalisationofNaV1.8innociceptor