ATP Enhances Spontaneous Calcium Activity in Cultured Suburothelial Myofibroblasts of the Human Bladder 英文参考文献.docVIP

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ATP Enhances Spontaneous Calcium Activity in Cultured Suburothelial Myofibroblasts of the Human Bladder 英文参考文献.doc

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ATP Enhances Spontaneous Calcium Activity in Cultured Suburothelial Myofibroblasts of the Human Bladder 英文参考文献

ATPEnhancesSpontaneousCalciumActivityinCultured SuburothelialMyofibroblastsoftheHumanBladder ShengCheng1,2.,FrankPeterScigalla1.,PietroSperonidiFenizio3,ZhiGengZhang2 ,Jens-Uwe Stolzenburg1,JochenNeuhaus1* 1Department of Urology, University of Leipzig, Leipzig, Germany, 2Department of Urology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou,China,3DepartmentofInformaticsEngineering,CISUC,UniversityofCoimbra,Coimbra,Portugal Abstract Background: Suburothelial myofibroblasts (sMF) are located underneath the urothelium in close proximity to afferent nerves.TheyexpresspurinergicreceptorsandshowcalciumtransientsinresponsetoATP.Thereforetheyaresupposedto beinvolvedinafferentsignalingofthebladderfullness.SinceATPconcentrationislikelytobeverylowduringtheinitial 2+ fillingphase,wehypothesizedthatsMFCa activityisaffectedevenatverylowATPconcentrations.WeinvestigatedATP inducedmodulationofspontaneousactivity,intracellularcalciumresponseandpurinergicsignalinginculturedsMF. Methodology/Principal Findings: Myofibroblast cultures, established from cystectomies, were challenged by exogenous ATPinpresenceorabsenceofpurinergicantagonist.Fura-2calciumimagingwasusedtomonitorATP(10216to1024 mol/l) induced alterations of calcium activity. Purinergic receptors (P2X1, P2X2, P2X3) were analysed by confocal immunofluorescence. We found spontaneous calcium activity in 55.18%61.65 of the sMF (N=48 experiments). ATP significantly increased calcium activity even at 10216mol/l. The calcium transients were partially attenuated by subtype selective antagonist (TNP-ATP, 1mM; A-317491, 1mM), and were mimicked by the P2X1, P2X3 selective agonist a,b- methyleneATP.TheexpressionofpurinergicreceptorsubtypesinsMFwasconfirmedbyimmunofluorescence. Conclusions/Significance:OurexperimentsdemonstrateforthefirsttimethatATPcanmodulatespontaneousactivityand induceintracellularCa responseinculturedsMFatverylowconcentrations,mostlikelyinvolvingP2Xreceptors.These 2+ findings