Electrogenic sodium-calcium exchange in cultured embryonic chick heart cells

1. The membrane potential (Em) of cultured chick embryonic heart cells depolarized to -36 mV after inhibition of the Na+-K+ pump by 0.1 mM-ouabain in a [K+]o of 24 mM: this was accompanied by a rise in Na+ content of approximately 65% in 3 min. Lowering [Na+]o to 27 mM then caused a fall in Na+ content, a rise in Ca2+ content and a small hyperpolarization of approximately 5 mV. The fall in Na+ content indicated a movement of Na+ which was in the opposite direction to the Na+ electrochemical gradient (a countergradient movement). 2. In the presence of 10 mM-Cs+ or 1 mM-Ba2+ the hyperpolarization was approximately 10 or approximately 30 mV, respectively. A 30 mV hyperpolarization took Em negative to the reversal potentials for K+, and Cl- as measured by ion-selective micro-electrodes. 3. The decay of the intracellular Na+ activity alpha iNa, in an [Na+]o of 27 mM followed a simple exponential time course (time constant, 36 s). The initial rate depended on the value to which [Na+]o was lowered in a manner suggesting a simple competitive inhibition of the exchange by external Na+. 4. The low-[Na+]o hyperpolarization was unaffected by amiloride (0.1 or 1 mM) or verapamil (20 microM). Both La3+ (1 mM) and Mn2+ (20 mM) blocked the hyperpolarization sufficiently to prevent Em hyperpolarizing negative to the reversal potentials for K+, Na+ and Cl-. 5. Re-establishing [Na+]o caused a rise in Na+ content and a countergradient drop in Ca2+ content. The effects of verapamil (20 microM), amiloride (0.1 and 1 mM), dichlorobenzamil (0.1 mM), quinidine (1 mM), Mn2+ (20 mM) and La3+ (1 mM) were tested on the movements of Na+ and Ca2+ both during exposure to an [Na+]o of 27 mM and on re-establishing [Na+]o. The only consistent and substantial effects were the attenuation by La3+ and Mn2+ and Ca2+ movements during exposure to an [Na+]o of 27 mM. However, neither La3+ nor Mn2+ affected the movements of Na+ and Ca2+ on re-establishing [Na+]o. 6. We conclude that cultured embryonic chick heart cells contain a Na+-Ca2+ exchange evidenced by the ability to cause movements of Na+ and Ca2+ which are counter to their respective electrochemical gradient and which are accompanied by downhill movements of the counter ion.(ABSTRACT TRUNCATED AT 400 WORDS)