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Dipartimento di Biologia Cellulare e Molecolare, Università di Perugia, I-06123 Perugia, Italy
Submitted 28 March 2003; accepted in final form 4 September 2003
A biophysical analysis of the voltage-gated K (Kv) currents of frog saccular hair cells enzymatically isolated with bacterial protease VIII was carried out, and their contribution to the cell electrical response was addressed by a modeling approach. Based on steady-state and kinetic properties of inactivation, two distinct Kv currents were found: a fast inactivating IA and a delayed rectifier IDRK. IA exhibited a strongly hyperpolarized inactivation V1/2 (-83 mV), a relatively rapid single exponential recovery from inactivation (
rec of
100 ms at -100 mV), and fast activation and deactivation kinetics. IDRK showed instead a less-hyperpolarized inactivation V1/2 (-48 mV), a slower, double-exponential recovery from inactivation (
rec1
490 ms and
rec2
4,960 ms at -100 mV), and slower activation and deactivation kinetics. Steady-state activation gave a V1/2 and a k of -46.2 and 8.2 mV for IA and -48.3 and 4.2 mV for IDRK. Both currents were not appreciably blocked by bath application of 10 mM TEA, but were inhibited by 4-AP, with IDRK displaying a higher sensitivity. IDRK also showed a relatively low affinity to linopirdine, being half blocked at
50 µM. Steady-state and kinetic properties of IDRK and IA were described by 2nd- and 3rd-order HodgkinHuxley models, respectively. The goodness of our quantitative description of the Kv currents was validated by including IA and IDRK in a theoretical model of saccular hair cell electrical activity and by comparing the simulated responses with those obtained experimentally. This thorough description of the IDRK and IA will contribute toward understanding the role of these currents in the electrical response on this preparation.
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