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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

The role of inactivation in the effects of n-alkanols on the sodium current of cultured rat sensory neurones.

1. The whole-cell patch-clamp technique has been used to investigate the actions of n-butanol, n-pentanol, n-hexanol and n-octanol on the sodium current of cells isolated from the dorsal root ganglia (DRGs) of neonatal rats and maintained in short-term tissue culture. 2. The influence of n-alkanols on the level of steady-state inactivation of the sodium current was investigated by a standard two-pulse protocol. All alkanols increased the level of resting inactivation and this was manifested as a hyperpolarizing shift of the relationship between the steady-state inactivation parameter (h infinity) and membrane potential. The mid-point of the h infinity curve was moved by up to -30 mV. 3. The relationship between the shift in the mid-point of the inactivation curve (delta Vh) and aqueous n-alkanol concentration has been derived for each n-alkanol. These are complex in shape and do not appear consistent with a hypothesis that the increase in inactivation results from 1:1 binding of an alkanol molecule to a single site on the channel protein. 4. The aqueous concentrations used ranged from 70 mM-n-butanol to 0.05 mM-n-octanol. However, equal fractional saturations of n-alkanols produced approximately equal shifts in the h infinity curve, particularly in the range 0.01-0.07 saturated. This implies a hydrophobic site of action, with a standard free energy per methylene group for adsorption to the site from the aqueous phase of ca -3.2 kJ/mol. 5. The increase in resting inactivation was not the sole means by which n-alkanols reduced the sodium current. The current was still reduced in cells pre-pulsed to sufficiently negative potentials to remove steady-state inactivation even in the presence of alkanols. The concentration required to reduce the current by 50% (ED50) has been interpolated for each n-alkanol. From these data it was estimated that the standard free energy per methylene group for adsorption to the site of action was ca -3.1 kJ/mol, similar to that calculated for the effect on inactivation. The concentration dependence of this residual block indicated the involvement of more than one n-alkanol molecule. 6. The n-alkanols increase the level of inactivation of rat DRG cell sodium channels at potentials around the resting membrane potential and this effect contributes to their local anaesthetic action.(ABSTRACT TRUNCATED AT 400 WORDS)[1]


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