This paper presents a study of the influence of isoproterenol (1 μM) on the force of isometric contractions (0.1–1.0 Hz; 30 ± 1°C; 1.8 mM Ca²⁺) of papillary muscles of the right ventricle in the heart of a ground squirrel during summer activity (n = 5) and hibernation season (activity between hibernation bouts, n = 4; torpor, n = 4; and arousal, n = 5). It is shown that isoproterenol increases the force of contraction (a positive inotropic effect) by 20 ± 3% and 61 ± 7% at stimulation frequencies of 0.4 and 1.0 Hz, respectively. In animals of hibernating period the isoproterenol-induced increase in the force of contraction is rather brief (within 3 min after onset of the influence) and is accompanied by a 30–50% decrease in the force from the control level (a negative inotropic effect) at stimulation frequencies from 0.3 to 0.8 Hz. The positive isoproterenol inotropic effect in active summer ground squirrels is associated with a decrease in a relative value of the pause potentiating effect (a qualitative indicator of calcium content in sarcoplasmic reticulum), and the negative inotropic effect, with its increase. In all groups of animals under examination the isoproterenol inotropic effect (regardless of its direction) is accompanied by the acceleration of the temporal parameters of the contraction—relaxation cycle. The dependence of isoproterenol effects in the heart of hibernating animals on both seasonal changes in calcium homeostasis and the activity of the sympathetic nervous system is under discussion.