Thyroid hormone deficiency is associated with changes in the cardiovascular system. No one has reported the measures of both atrial and ventricular calcium channel density and function, in association with measures of negative and positive channel modulators, in hypothyroid hearts. Hormonally-induced modulation of calcium channels has clinical significance in the development and application of therapeutic agents in dysthyroid states. We thyroidectomized male rats and sham-operated euthyroid controls, in order to measure radioligand binding to ventricular and atrial membrane Ca2+ channels ([3H]-isradipine), beta-adrenoceptors ([125I]-iodocyanopindolol) and atrial muscarinic receptors ([3H]-quinuclidinyl benzilate) and related these data to contraction and heart rate responses to isoproterenol, carbachol and calcium. When data from hypothyroid tissues were compared to those of controls, the densities of calcium channels increased 50% in ventricles, but no differences were seen in atrial homogenates. In both atria and ventricles, beta-adrenoceptors decreased modestly with no change in affinity. Atrial muscarinic receptor density was also unchanged. Dose response curves of left atrial contractions showed: decreased sensitivity (increased EC50 value) but equal maximal responsiveness to extracellular calcium; an increased carbachol sensitivity (decreased EC50 value); and no significant difference in isoproterenol response. Comparisons of within-individual preparation ratioed EC50 values confirm the changed calcium and carbachol sensitivities. Heart-rate dose response curves displayed: increased maximal heart rate responsiveness to calcium associated with increased EC50 values; isoproterenol sensitivity was decreased nearly 3-fold. There was no significant difference in heart rate response to carbachol; however, ratioed values of carbachol and calcium EC50s were significantly different. These data are consistent with a tissue-level state of enhanced negative chronotropism and inotropism occurring in hypothyroid myocardia. We also confirm an earlier controversial finding of increased calcium channel density in ventricles from hypothyroid rats.