Nonuniform responses of transmembrane potential during electric field stimulation of single cardiac cells

The response of cellular transmembrane potentials (V(m)) to applied electric fields is a critical factor during electrical pacing, cardioversion, and defibrillation, yet the coupling relationship of the cellular response to field intensity and polarity is not well documented. Isolated guinea pig ventricular myocytes were stained with a voltage-sensitive fluorescent dye, di-8-ANEPPS (10 microM). A green helium-neon laser was used to excite the fluorescent dye with a 15-micrometers-diameter focused spot, and subcellular V(m) were recorded optically during field stimulation directed along the long axis of the cell. The membrane response was measured at the cell end with the use of a 30-ms S1-S2 coupling interval and a 10-ms S2 pulse with strength of up to approximately 500-mV half-cell length potential (field strength x one-half the cell length). The general trends show that 1) the response of V(m) at the cell end occurs in two stages, the first being very rapid (<1 ms) and the second much slower in time scale, 2) the rapid response consists of hyperpolarization when the cell end faces the anode and depolarization when the cell end faces the cathode, 3) the rapid response varies nonlinearly with field strengths and polarity, being relatively larger for the hyperpolarizing responses, and 4) the slower, time-dependent response has a time course that varies in slope with field strength. Furthermore, the linearity of the dye response was confirmed over a voltage range of -280 to +140 mV by simultaneous measurements of optically and electrically recorded V(m). These experimental findings could not be reproduced by the updated, Luo-Rudy dynamic model but could be explained with the addition of two currents that activate outside the physiological range of voltages: a hypothetical outward current that activates strongly at positive potentials and a second current that represents electroporation of the cell membrane.