膜片钳技术的发展及其应用

第27卷　第2期　　运城学院学报Vol.27　No.22009年4月　JournalofYunchengUniversityApr.2009膜片钳技术的发展及其应用曹建斌＊(,044000)　　　:,。,,、、、。,,。:;;;:Q424　:A　　:1008-8008(2009)02-0053-03　　。,,、。,。,HuxleyKatz[1],,,。。1.(patch-clamptechnique),,()。,,(),(1)。1976,NeherSakmann[2],(10-12A)。1981,HamillNeher[3],1pA,10μs1μm,1　[4]。,:、、、[4](2)。2　[4]2.[5]2.1根据实验的需要,拉制微电极,,200-400μm,,,1-2μm。·53·＊:2009-02-16:(1978-),,,,,。DOI:10.15967/j.cnki.cn14-1316/g4.2009.02.015,,,,,。,,,。2.2充灌微电极,。,,,。,,,,,,。,,,。2.3高阻封接的形成,,,,。,,,,,。2.4全细胞记录模式的形成,,(ZAP),,C-Fast、τ-FastC-Slow、τ-Slow,。,,,。3.1976NeherSakmann,,,。3.1膜片钳技术与荧光技术的结合Fura-2[6-7]、Fluo-3[8-9]、Fluo-4[10],,,,,,、。3.2穿孔膜片钳技术1988Horn[11],(perforatedpatchclamptechnique):,,。,0.4～0.8nm,Na+、K+、Li+、Cs+、Cl-,,。,,[12]。,4℃。,,。,,。3.3在体膜片钳技术。,2090,Pei[13],。,Blan-ton[14]Margrie[15]“”,,。Margrie[16],TPTP(two-photontargetedpatching)。,,,、[17],。3.4全自动膜片钳技术,、[18]。,,,,,。SophionBiosci-enceQPatch96,,[19-20]。,,[21]。3.5膜片钳技术与分子生物学技术的结合,,,。,mRNA。,,mRNAcDNA,PCR,PCRDNA[22]。,,,。·54·4.,。,、,、、、、、、、。,,,。,,50μm[23]。,,、,,[24]。,。:[1]Hodgkin,AL,Huxley,AFandKatz,B.Measurementofcurrent-voltagerelationsinthemembraneofthegiantax-onofLoligo[J].JPhysiol.1952,116(4):424-448.[2]Neher,EandSakmann,B.Single-channelcurrentsre-cordedfrommembraneofdenervatedfrogmusclefibres[J].Nature.1976,260(5554):799-802.[3]Hamill,OP,Marty,A,Neher,E,etal.Improvedpatch-clamptechniquesforhigh-resolutioncurrentrecordingfromcellsandcell-freemembranepatches[J].PflugersArch.1981,391(2):85-100.[4].[M].:,2001.[5].[D].:,2006.[6]Callewaert,G,Cleemann,LandMorad,M.EpinephrineenhancesCa2+current-regulatedCa2+releaseandCa2+reuptakeinratventricularmyocytes[J].ProcNatlAcadSciUSA.1988,85(6):2009-2013.[7]Das,A,Garner,DP,DelRe,AM,etal.Calpeptinpro-videsfunctionalneuroprotectiontoratretinalganglioncellsfollowingCa2+influx[J].BrainRes.2006,1084(1):146-157.[8]Garcia,J,Tanabe,TandBeam,KG.Relationshipofcal-ciumtransientstocalciumcurrentsandchargemovementsinmyotubesexpressingskeletalandcardiacdihydropyridinereceptors[J].JGenPhysiol.1994,103(1):125-147.[9]Collier,ML,Thomas,APandBerlin,JR.RelationshipbetweenL-typeCa2+currentandunitarysarcoplasmicre-ticulumCa2+releaseeventsinratventricularmyocytes[J].JPhysiol.1999,516(Pt1):117-128.[10]Sheridan,DC,Carbonneau,L,Ahern,CA,etal.Ca2+-dependentexcitation-contractioncouplingtrig-geredbytheheterologouscardiac/brainDHPRbeta2a-subunitinskeletalmyotubes[J].BiophysJ.2003,85(6):3739-3757.[11]Horn,RandMarty,A.Muscarinicactivationofioniccurrentsmeasuredbyanewwhole-cellrecordingmethod[J].JGenPhysiol.1988,92:145-159.[12],,,.[J]..2006,28(2):62-65.[13]Pei,X,Volgushev,M,Vidyasagar,TR,etal.Wholecellrecordingandconductancemeasurementsincatvisualcortexin-vivo[J].Neuroreport.1991,2(8):485-488.[14]Blanton,MG,LoTurco,JJandKriegstein,AR.Wholecellrecordingfromneuronsinslicesofreptilianandmammaliancerebralcortex[J].JNeurosciMethods.1989,30(3):203-210.[15]Margrie,TW,Sakmann,BandUrban,NN.Actionpo-tentialpropagationinmitralcelllateraldendritesisdecre-mentalandcontrolsrecurrentandlateralinhibitioninthemammalianolfactorybulb[J].ProcNatlAcadSciUSA.2001,98(1):319-324.[16]Margrie,TW,Meyer,AH,Caputi,A,etal.Targetedwhole-cellrecordingsinthemammalianbraininvivo[J].Neuron.2003,39(6):911-918.[17],,,.[J]..2005,28(2):91-92.[18],,,.-[J]..2007(2):47-50.[19]Asmild,M,Oswald,N,Krzywkowski,KM,etal.Up-scalingandautomationofelectrophysiology:towardhighthroughputscreeninginionchanneldrugdiscovery[J].ReceptorsChannels.2003,9(1):49-58.[20]Mathes,C.QPatch:thepast,presentandfutureofauto-matedpatchclamp[J].ExpertOpinTherTargets.2006,10(2):319-327.[21]Brown,AM.Highthroughputfunctionalscreeningofanionchannellibraryfordrugsafetyandefficacy[J].EurBiophysJ.2008.[22],,,.[J]..2001,8(2):24-26.[23],,,.[J].().2006,25(5):100-103.[24],,,.-[J]..2008,27(2):127-130.【　　】·55·