Interpretation of [3H]ouabain binding in guinea-pig ventricular myocardium in relation to sodium pump activity.
AUTOR(ES)
Herzig, S
RESUMO
1. The attempt was made to analyse the complex [3H]ouabain binding curves obtained in intact cardiac ventricular preparations electrically stimulated at different frequencies. The result of this analysis was used to draw conclusions from the binding curves on the frequency dependence of sodium pump activity. 2. [3H]Ouabain binding to isolated, electrically stimulated (1.5 Hz) ventricular strips of guinea-pig hearts was investigated. The positive inotropic effects were studied in separate experiments. Specific [3H]ouabain binding barely reached an equilibrium within 3 h of incubation. A binding curve was constructed using the equilibrium values of specific [3H]ouabain binding obtained at different ouabain concentrations. This binding curve revealed a concentration-proportional component at positive inotropic concentrations and a saturating component at high, toxic concentrations. At very low, inotropically ineffective ouabain concentrations, however, binding values were higher than expected from a linear relationship between ouabain concentration and binding. 3. The peculiar shape of the binding curve could be largely accounted for by a mathematical model, which takes into consideration biochemical properties and physiological regulation of the sodium pump. The model predicts a concentration-proportional pattern of binding which takes place in the non-toxic ouabain concentration range. The slope of the concentration-proportional component of the binding curve should represent a measure of sodium pump activity. 4. Investigation of binding curves at various stimulation frequencies revealed that, as predicted by the model, the slope of the concentration-proportional component of the binding curves was increased and the maximum non-toxic equilibrium binding was decreased with increasing beat frequencies. 5. Quantitative evaluation of the binding curves led to the conclusion that sodium pump activity is a linear function of stimulation frequency in guinea-pig ventricular preparations, the activity in resting preparations amounting to about 15% of the maximum activity. Comparison of the present results with former studies on sodium pump function suggests that [3H]ouabain binding reflects steady-state sodium pump activity. If the complex pattern of binding curves is taken into consideration, [3H]ouabain binding measurements may serve as a means of studying sodium pump function.
ACESSO AO ARTIGO
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