Beat-to-Beat Variation in Periodicity of Local Calcium Releases Contributes to Intrinsic Variations of Spontaneous Cycle Length in Isolated Single Sinoatrial Node Cells.docVIP

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Beat-to-Beat Variation in Periodicity of Local Calcium Releases Contributes to Intrinsic Variations of Spontaneous Cycle Length in Isolated Single Sinoatrial Node Cells.doc

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Beat-to-Beat Variation in Periodicity of Local Calcium Releases Contributes to Intrinsic Variations of Spontaneous Cycle Length in Isolated Single Sinoatrial Node Cells

Beat-to-BeatVariationinPeriodicityofLocalCalcium ReleasesContributestoIntrinsicVariationsof SpontaneousCycleLengthinIsolatedSingleSinoatrial NodeCells OliverMonfredi1,2,LarissaA.Maltseva1,HaroldA.Spurgeon1,MarkR.Boyett2,EdwardG.Lakatta1, VictorA.Maltsev1* 1LaboratoryofCardiovascularScience,NationalInstituteonAging-IntramuralResearchProgram,NationalInstitutesofHealth,Baltimore,Maryland,UnitedStatesof America,2InstituteofCardiovascularSciences,UniversityofManchester,Manchester,UnitedKingdom Abstract Spontaneous,submembranelocalCa2+releases(LCRs)generatedbythesarcoplasmicreticuluminsinoatrialnodalcells,the cellsoftheprimarycardiacpacemaker,activateinwardNa /Ca -exchangecurrenttoacceleratethediastolicdepolarization + 2+ 2+ rate,andthereforetoimpactoncyclelength.SinceLCRsaregeneratedbyCa releasechannel(i.e.ryanodinereceptor) openings,theyexhibitadegreeofstochasticbehavior,manifestedasnotablecycle-to-cyclevariationsinthetimeoftheir occurrence. Aim: The present study tested whether variation in LCR periodicity contributes to intrinsic (beat-to-beat) cycle length variabilityinsinglesinoatrialnodalcells. Methods: We imaged single rabbit sinoatrial nodal cells using a 2D-camera to capture LCRs over the entire cell, and, in selectedcells,simultaneouslymeasuredactionpotentialsbyperforatedpatchclamp. Results:LCRsbegintooccuronthedescendingpartoftheactionpotential-inducedwhole-cellCa2+transient,ataboutthe time of the maximum diastolic potential. Shortly after the maximum diastolic potential (mean 5467.7ms, n=14), the 2+ ensembleofwaxingLCRactivityconvertsthedecayoftheglobalCa transientintoarise,resultinginalate,whole-cell diastolic Ca2+ elevation, accompanied by a notable acceleration in diastolic depolarization rate. On average, cells (n=9) generate13.263.7LCRspercycle(mean6SEM),varyinginsize(7.164.2mm)andduration(44.2627.1ms),withbothsize anddurationbeinggreaterforlater-occurringLCRs.WhilethetimingofeachLCRoccurrencealsovaries,theLCRperiod(i.e. 2+ thetimefro