Under tissue bath conditions, isolated specimens of human right atrium are characterized by the presence of large numbers of partially depolarized cells. The basis for the depolarization is still not understood. To determine if reduced intracellular potassium activity (aKi) is responsible for the low levels of maximum diastolic potential (MDP), aKi was directly measured with potassium ion-selective micro-electrodes (KISE). The effects of varying bath potassium concentration ([K+]0) on aiK and on the relationship between MDP and the potassium equilibrium potential (EK) also were determined. In 4 mM [K+]0, all specimens studied were partially depolarized (MDP = -43.9 +/- 1.3 mV [mean +/- S.E.]) and beat spontaneously. aKi was 98.1 +/- 1.5 mM, and EK was -93.3 +/- 0.4 mV. Changes in [K+]0 between 2.5 and 40 mM did not change aKi significantly. MDP was relatively insensitive to [K+]0 and was always far positive to EK over the entire range of [K+]0 studied. The ability of the KISE to reliably measure aKi in the face of the spontaneous diastolic depolarization was assessed by using acetylcholine or cooling to depress automaticity. These measures did not significantly alter the calculated aKi. The results indicate that: aKi in human atrial specimens is similar to that in atria from other mammalian species, and the low level of MDP exhibited by human atrial cells subject to conventional tissue bath conditions does not primarily result from low levels of aKi.