Ionic diffusion in transverse tubules of cardiac ventricular myocytes

We have estimated the rate of diffusion of calcium ions in the transverse tubules of isolated cardiocytes by recording changes in peak calcium current (ICa) caused by rapid changes of the extracellular calcium concentration ([Ca]o) at various intervals just preceding activation of ICa. Isolated ventricular cells of guinea pig heart and atrial cells from rabbit heart were voltage-clamped (whole cell patch), superfused at a high flow rate, and stimulated continuously with depolarizing pulses (0.5 Hz, 200- or 20-ms pulses from a holding potential of -45 or -75 mV to 0 mV). In ventricular cells, the change in peak ICa following a sudden change of [Ca]o increased rapidly as the delay between the solution change and depolarization was increased, up to a delay of approximately 75 ms [time constant (tau) approximately 20 ms, 30-40% of total current change), and then increased more slowly (tau approximately 200 ms, 60-70% of total current change); 400-500 ms were needed to achieve 90% of the total current increase. In atrial cells, a clear separation into two phases was not possible and 90% of the current change occurred within 85 ms. The slow phase of current change, which was unique to the ventricular cells, presumably reflects the slow equilibration of ions between the bulk perfusate and the lumina of the transverse tubules. If the lengths of the transverse tubules were equal to the cell thickness, the slow rate of change of current would be consistent with an apparent diffusion coefficient for calcium ions of 0.95 x 10(-6) cm2/s, considerably smaller than the value in bulk solution (7.9 x 10(-6) cm2/s). Most likely, this discrepancy is due to a high degree of tortuosity in the transverse tubular system in guinea pig ventricular cells or possibly to ion binding sites within the tubular membranes and glycocalyx.