The effects of omission of buffers from Krebs-Ringer medium on contractile activity, membrane potentials and ATP content of electrically stimulated isolated rat atria were investigated. 1) Contractile status: A rapid and marked depression of the contractile activity of atria occurred when buffer-free medium was substituted for the normal Krebs-Ringer medium. 2) Electrical status: The omission of buffers from medium did not alter the resting or action potential magnitudes of atria. However, the action potential duration was on initial increase followed by a decrease in the buffer-free medium. 3) ATP concentration: The omission of buffers from medium resulted in a marked decrease in the ATP levels of atria. It has been also found in the present study that bicarbonate buffer plays an important role for the maintenance of the contractility and ATP levels of the heart. The contractile depression by the omission of buffers was not directly associated with electrical alterations in resting or action potentials of the heart. In the absence of bicarbonate-buffer, glucose no longer plays to maintain the contractile activity and the ATP levels of rat atria.