Mitochondrial Ca2+ (Ca2+m) plays a fundamental role in the cardiac excitation-contraction process. Cytosolic Ca2+ (Ca2+c) enters mitochondria through the mitochondrial uniporter and is extruded by the mitochondrial Na+-Ca2+ exchanger. Mitochondria buffer Ca2+c during normal and pathological conditions. For example an increase in ventricular cardiomyocyte workload elevates the cytosolic Ca2+ concentration (Ca2+c), which effects Ca2+m concentration (Ca2+m), permitting Ca2+-dependent activation of mitochondrial enzymes to match ATP production to the increase in ATP demand. Under pathological conditions Ca2+c overload may cause Ca2+m overload, promoting cell injury. In contrast, Ca2+m buffering, up to a point, counteracts cardiac glycoside toxicity derived from Ca2+c overload. Normal automaticity in sinoatrial node cells (SANC) involves Ca2+c cycling within a coupled-clock system: periodic local, subsarcolemmal Ca2+ releases (LCRs) from sarcoplasmic reticulum (Ca2+ clock) activate an inward Na+-Ca2+ exchange current that accelerates the diastolic depolarization prompting the ensemble of surface membrane ion channels (membrane clock) to generate the next action potential (AP). We hypothesized that dynamic Ca2+m cycling impacts on SANC normal automaticity. Specific inhibitors of Ca2+ influx into (Ru360) and Ca2+ efflux from (CGP-37157) mitochondria in intact single isolated SANC were used. Ca2+m was indexed by selectively quenching the cytosolic fluorescence Ca2+ probe, Indo-1 by Mn2+. Ru360 decreased Ca2+m over 10-min and increased SANC spontaneous AP firing rate. Conversely, CGP-37157 increased Ca2+m over 10-min and reduced AP firing rate. Inhibition of Ca2+m influx significantly increased the SR Ca2+ load, LCR size, duration and amplitude (imaged via confocal linescan), and shortened the LCR period. Conversely, inhibition of Ca2+ efflux from mitochondria decreased the SR Ca2+ load, LCR characteristics, and lengthened the LCR period. Changes in total Ca2+ released by the LCR ensemble were highly correlated with the change in the SR Ca2+ load (r2=0.97). Changes in LCR period induced by changes in Ca2+m flux predicted (r2=0.84) concurrent changes in the spontaneous SANC AP cycle length. Our novel numerical model simulations reproduce our experimental findings and validate our interpretation of these findings: changes in mitochondrial Ca2+ flux affect the spontaneous AP firing rate of SANC via changes in SR Ca2+ loading and release. Furthermore, while our experimental method to measure Ca2+m does not permit evaluation of systolic and diastolic Ca2+m, our model predicts that under basal conditions both diastolic and systolic Ca2+m are lower than the diastolic and systolic Ca2+c, respectively. However, the systolic Ca2+m is higher than diastolic Ca2+c. In summary, A change in SANC Ca2+m flux translates into a change in the AP firing rate by effecting changes in Ca2+c and SR Ca2+ loading, which affects the characteristics of spontaneous SR Ca2+ release.
|Yaniv, Yael; Ahmet, Ismayil; Liu, Jie et al. (2014) Synchronization of sinoatrial node pacemaker cell clocks and its autonomic modulation impart complexity to heart beating intervals. Heart Rhythm 11:1210-9|
|Liu, Jie; Sirenko, Syevda; Juhaszova, Magdalena et al. (2014) Age-associated abnormalities of intrinsic automaticity of sinoatrial nodal cells are linked to deficient cAMP-PKA-Ca(2+) signaling. Am J Physiol Heart Circ Physiol 306:H1385-97|
|Maltsev, Victor A; Yaniv, Yael; Maltsev, Anna V et al. (2014) Modern perspectives on numerical modeling of cardiac pacemaker cell. J Pharmacol Sci 125:6-38|
|Yaniv, Yael; Stern, Michael D; Lakatta, Edward G et al. (2013) Mechanisms of beat-to-beat regulation of cardiac pacemaker cell function by Ca²? cycling dynamics. Biophys J 105:1551-61|
|Lakatta, Edward G; Yaniv, Yael; Maltsev, Victor A (2013) Minding the gaps that link intrinsic circadian clock within the heart to its intrinsic ultradian pacemaker clocks. Focus on ""The cardiomyocyte molecular clock, regulation of Scn5a, and arrhythmia susceptibility"". Am J Physiol Cell Physiol 304:C941-4|
|Yaniv, Yael; Lakatta, Edward G (2013) Pacemaker gene mutations, bradycardia, arrhythmias and the coupled clock theory. J Cardiovasc Electrophysiol 24:E28-9|
|Yaniv, Yael; Spurgeon, Harold A; Lyashkov, Alexey E et al. (2012) Crosstalk between mitochondrial and sarcoplasmic reticulum Ca2+ cycling modulates cardiac pacemaker cell automaticity. PLoS One 7:e37582|