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Department of Neurophysiology, Tohoku University School of Medicine, Sendai, Japan.
1. Tetrodotoxin (TTX)-sensitive Ca2+ conducting channels which produce a transient inward current were investigated in pyramidal neurones freshly dissociated from the dorsal part of rat hippocampal CA1 region by the use of the suction-pipette technique, which allows for intracellular perfusion under a single-electrode voltage clamp. 2. In all cells superfused with Na(+)- and K(+)-free external solution containing 10 mM-Ca2+ and 10(-5) M-La3+, a transient inward Ca2+ current was evoked by a step depolarization to potentials more positive than about -50 mV from a holding potential (VH) of -100 mV. This current was inhibited by either removing the extracellular Ca2+ or adding TTX (termed as 'TTX-ICa'). 3. Activation and inactivation processes of the TTX-ICa were highly potential dependent at 20-22 degrees C, and the latter was fitted by a double exponential function. The time to peak of the current decreased from 5.0 to 2.3 ms at a test potential change from -50 to 0 mV. The time constants of the current decay decreased from 2.8 to 2.2 ms for fast component (tau if) and from 16.0 to 8.2 ms for slow component (tau is) at a potential change from -35 to -10 mV. 4. The TTX-ICa was activated at threshold potential of about -55 mV and reached full activation at -30 mV. The steady-state inactivation of TTX-ICa could be fitted by a Boltzmann equation with a slope factor of 6.0 mV and a half-inactivation voltage of -72.5 mV. 5. Biphasic recovery (reactivation) from the complete inactivation of TTX-ICa was observed. The time constant of the major component (78.8 to 91.6% of total) of the reactivation was 13.1 ms, and that of the minor one was 120 to 240 ms. Therefore, TTX-ICa remained fairly constant at a train of stimulation up to 3 Hz. However, the inhibition of current amplitude occurred as the repetitive stimulation increased more than 10 Hz, and considerable tonic inhibition occurred with increasing stimulation frequency. 6. When the peak amplitudes in the individual current-voltage (I-V) relationships of TTX-ICa at various extracellular Ca2+ concentrations ([Ca2+]o) were plotted as a function of [Ca2+]o, the current amplitude increased linearly without showing any saturation. 7. The ratio of peak amplitude in the individual I-V relationships of Ca2+, Sr2+ and Ba2+ currents passing through the TTX-sensitive Ca2+ conducting channel was 1:0.33:0.05, although the current kinetics were much the same.(ABSTRACT TRUNCATED AT 400 WORDS)
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