Bistramide A, a new toxin isolated from the Urochordate Lissoclinum bistratum Sluiter, was applied to rat auricular heart muscle bundles. At a stimulation frequency of 0.2 Hz, the toxin induces a dose-dependent reduction of the stimulated twitch tension force; it decreases Vmax and shortens the duration of the plateau and the slow repolarizing phase of the action potential. In the control solution, switching from a stimulation frequency of 0.2 Hz to 1 Hz decreases the force with which a positive potentiation develops either at a maintained high frequency or after switching from 1 Hz to 0.2 Hz. Bistramide A reduces both the force evoked at 1 Hz and the potentiation. The data suggest that Bistramide A blocks Na+ conductance; inhibits Ca++ channels in a time- and frequency-dependent manner; reduces Na(+)-Ca++ exchange activity; but does not modify the ability of the sarcoplasmic reticulum to be refilled although the rate of Ca++ accumulation is decreased.