Ionic basis of ischemia-induced bradycardia in the rabbit sinoatrial node

Abstract 

To investigate the basis of ischemia-induced bradycardia (<60 beats/min), we isolated pacemaker cells from the rabbit sinoatrial node and exposed them to ischemic-like conditions, including omission of glucose, pH 6.6, and either 5.4 or 10 mM KCl to evaluate the role of increased serum [K]. A perforated-patch technique was employed to test the hypothesis that the arrhythmia is caused by attenuation of inward currents that contribute to the diastolic depolarization. After exposure to “ischemic” Tyrode containing 5.4 mM KCl, the pacemaker cells exhibited 13% slower beat rates and action potentials with 6-mV greater overshoots and 44% longer durations. In contrast, after exposure to “ischemic” Tyrode containing 10 mM KCl, the pacemaker cells exhibited a 7-mV depolarization of the maximum diastolic potential but no significant change in the overshoot. Beat rates were slowed by 43%, and the action potentials were prolonged by 46%. “Ischemic” Tyrode containing 5.4 mM KCl increased L-type Ca current, decreased T-type Ca current and reduced Ni-sensitive inward current tails (presumably Na–Ca exchange current), even after treatment with 40 μM ryanodine to block Ca release from the sarcoplasmic reticulum. “Ischemic” Tyrode containing 10 mM KCl increased hyperpolarization-activated inward current at diastolic potentials and reduced the slowly activating component, but not the rapidly activating component, of delayed rectifier K current. Our results suggest that reductions of inward Na–Ca exchange current and T-type Ca current contribute to “ischemia”-induced “bradycardia” in sinoatrial node pacemaker cells.

Abbreviations: SAN, sinoatrial node, PCs, pacemaker cells, BR, beat rate, AP, action potential, MDP, maximum diastolic potential, OS, overshoot, DUR, duration, I_f, hyperpolarization-activated inward current, I_Ca,L, L-type Ca current, I_Ca,T, T-type Ca current, I_NCX, Na–Ca exchange current, I_K, delayed rectifier K current, I_K,r, rapidly activating component of delayed rectifier K current, I_K,s, slowly activating component of delayed rectifier K current, I_K(ATP), ATP-sensitive K current, SR, sarcoplasmic reticulum, E_rev, reversal potential, NaCN, Na cyanide, Ca_i, intracellular Ca, CaMKII, Ca/calmodulin-dependent protein kinase II.

Keywords: Sinoatrial node, Bradycardia, Ischemia, Rabbit, L-type Ca current, T-type Ca current, Na–Ca exchange current, Hyperpolarization-activated inward current, Delayed rectifier K current