New Insights in the Contribution of Voltage-Gated Nav Channels to Rat Aorta Contraction 英文参考文献.docVIP

NewInsightsintheContributionofVoltage-GatedNav ChannelstoRatAortaContraction Aure′lieFort1,MagaliCordaillat1,CatherineThollon2,GuillermoSalazar1,IlanaMechaly3 ,Nicole Villeneuve2,Jean-PaulVilaine2,SylvainRichard1*,AnneVirsolvy1 1Inserm U637, Universite′ Montpellier1 2, Montpellier, France, 2Cardiovascular Division, Institut de Recherches Servier, Suresnes, France, 3Inserm U583, Universite′ Montpellier2,Montpellier,France Abstract Background:Despiteincreasingevidenceforthepresenceofvoltage-gatedNa+channels(Nav)isoformsandmeasurements of Nav channel currents with the patch-clamp technique in arterial myocytes, no information is available to date as to whetherornotNavchannelsplayafunctionalroleinarteries.Theaimofthepresentworkwastolookforaphysiological roleofNavchannelsinthecontrolofrataorticcontraction. Methodology/Principal Findings: Nav channels were detected in the aortic media by Western blot analysis and double immunofluorescencelabelingforNavchannelsandsmoothmusclea-actinusingspecificantibodies.Inparallel,usingreal timeRT-PCR,weidentifiedthreeNavtranscripts:Nav1.2,Nav1.3,andNav1.5.OnlytheNav1.2isoformwasfoundintheintact mediaandinfreshlyisolatedmyocytesexcludingcontaminationbyothercelltypes.UsingthespecificNavchannelagonist veratridine and antagonist tetrodotoxin (TTX), we unmasked a contribution of these channels in the response to the depolarizing agent KCl on rat aortic isometric tension recorded from endothelium-denuded aortic rings. Experimental conditions excluded a contribution of Nav channels from the perivascular sympathetic nerve terminals. Addition of low concentrations of KCl (2–10mM), which induced moderate membrane depolarization (e.g., from 255.961.4mV to 245.961.2mV at 10mmol/L as measured with microelectrodes), triggered a contraction potentiated by veratridine + 2+ (100mM)andblockedbyTTX(1mM).KB-R7943,aninhibitorofthereversemodeoftheNa /Ca exchanger,mimickedthe effectofTTXandhadnoadditiveeffectinpresenceofTTX. Conclusions/Significa