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1 Dipartimento di Biologia Cellulare e Molecolare, University of Perugia, Perugia, Italy
* To whom correspondence should be addressed. E-mail: fabiolab{at}unipg.it.
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 modelling 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 ~4960 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 proprerties of IDRK and IA were described by second and third order Hodgkin-Huxley 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 towards understanding the role of these currents in the electrical response on this preparation.
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