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Department of Pharmacology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.

1. The gating properties of two types of Na+ channels were studied in neurones isolated from rat dorsal root ganglia using the whole cell variation of the patch electrode voltage-clamp technique. 2. Two types of Na+ currents (INa) were identified on the basis of their sensitivity to tetrodotoxin (TTX). One type was insensitive to TTX (up to 0.1 mM), while the other type was blocked by 1 nM of TTX. Whereas they were both insensitive to 50 microM Cd2+, a high concentration (2 mM) of Co2+ selectively inhibited the TTX-insensitive type. 3. The activation thresholds were about -60 and -40 mV for the TTX-sensitive and the TTX-insensitive INa, respectively. Activation of the TTX-sensitive INa developed with a sigmoidal time course which was described by m3 kinetics, whereas the activation of the TTX-insensitive INa was described by a single exponential function. A deactivation process, as measured by the tail current upon repolarization, followed an exponential decay in either type of INa. 4. The rate constant of activation indicated that under comparable membrane potential conditions, the TTX-insensitive channels open 4-5 times slower than the TTX-sensitive ones upon depolarization. Likewise, the rate constant of inactivation indicated that the TTX-insensitive channels inactivate 3-7 times more slowly than the TTX-sensitive ones upon repolarization. 5. The steady-state activation curve for the TTX-insensitive INa was shifted about 20 mV in the positive direction from that for the TTX-sensitive INa. 6. The steady-state inactivation curve for the TTX-insensitive INa as obtained with a 0.5 s prepulse was shifted about 26 mV in the positive direction from that for the TTX-sensitive INa, indicating a greater availability for the TTX-insensitive INa in depolarized membrane. However, on increasing the duration of prepulse, the inactivation curve for the TTX-insensitive INa, but not for the TTX-sensitive INa, shifted in the negative direction due to an extremely slow inactivation process in the TTX-insensitive INa. Consequently, an overlap between the activation and inactivation curves which causes a steady influx of Na+ (window current) became progressively reduce. 7. The time course of INa decay was best described by a single exponential process in either the TTX-sensitive or TTX-insensitive INa, whereas the development of inactivation and the recovery from inactivation, which were measured by a conventional double-pulse protocol, followed a second order process in either channel type.(ABSTRACT TRUNCATED AT 400 WORDS)

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