Abstract. Sensitivity of cardiac muscle to digitalis glycosides depends on the thyroid state. The mechanism of this interaction was investigated at the cellular level using spontaneously beating monolayers of cultured chick embryo ventricular cells. Cells were grown for 48 h in serum-free medium containing concentrations of triiodothyronine (T3) from zero to 10-7 M, and the total number of sodium pump sites, sodium content, and contractile amplitude in the presence and absence of various concentrations of ouabain were determined. T3 caused a concentration-dependent increase in the number of specific ouabain binding sites; the maximal increase to 160% of control was observed in response to 10-8 M T3. T3 lowered steady-state cellular sodium content in a concentration-dependent manner, also. Ouabain (1 ,uM) exposure elevated cellular sodium content in all cells, but the increase was greatest in cells grown in T3-free medium and least in cells grown in 10-8 M T3. The positive inotropic and toxic effects of ouabain in cells grown in 10-8 M T3 were diminished at any given ouabain concentration, and thus, the dose-response curve was shifted to the right. These results indicate that T3 causes induction of additional sodium pump sites that are functional. The increased tolerance of hyperthyroid cells and reduced tolerance of hypothyroid cells to cardiac glycosides can be explained by these changes in the number of sodium pump sites and cellular sodium

We investigated the roles of sodium-calcium exchange, sarcoplasmic reticulum, and mitochondria in Cai homeostasis in cultured chick ventricular cells. Specifically, the influence of low sodium medium on contractile state, calcium fluxes, and cytosolic free [Ca] [( Ca]i) was examined. [Ca]i was measured using fura-2. Mean [Ca]i in control medium was 126 +/- 14 nM. Exposure of cells to sodium-free or sodium- and calcium-free medium (choline-substituted) resulted in contracture development, which returned toward the baseline level over 2-3 minutes. The Nao-free contracture was associated with a tenfold increase in [Ca]i (1,280 +/- 110 nM) followed by a gradual decrease to a level fourfold above control [Ca]i (460 +/- 58 nM). Nao- and Cao-free contracture was associated with a fivefold increase in [Ca]i (540 +/- 52 nM) followed by a rapid decrease to below 80 nM. Sodium-free medium failed to produce an increase in [Ca]i or contracture in cells preexposed to calcium-free medium, although caffeine, when subsequently added to sodium- and calcium-free medium, was able to elicit a transient increase in [Ca]i and contracture. Brief, 5-second preperfusion of cells with La3+ (1 mM) or EGTA (1 mM) abolished the Nao-free contracture and the increase in [Ca]i. In the presence of 20 mM caffeine, removal of Nao resulted in minimal changes in the resting position of the cell although 45Ca uptake and [Ca]i were increased in response to sodium-free medium; the subsequent decrease in [Ca]i was greatly slowed. Addition of caffeine during the relaxation phase of the sodium-free contracture produced an additional transient contracture and transient increase in [Ca]i. Ryanodine (1 microM) abolished this effect of caffeine. Caffeine or ryanodine abolished Nao- and Ca-free contracture. CCCP (2 microM), a potent oxidative phosphorylation inhibitor, did not significantly affect calcium efflux rate. In the presence of 2 microM CCCP, removal of sodium resulted in an augmented contracture signal and a rise in [Ca]i, followed by a slow decrease. We conclude that removal of extracellular sodium enhances transsarcolemmal entry of calcium via sodium-calcium exchange, but this effect alone does not lead to the development of sodium-free contracture. Calcium displaceable by lanthanum or EGTA appears to contribute to Nao-free or Nao- and Cao-free contracture. Studies using caffeine and ryanodine suggest that removal of Nao leads to release of calcium from the sarcoplasmic reticulum (presumably via calcium-induced calcium release).(ABSTRACT TRUNCATED AT 400 WORDS)