Cholinergic control of membrane conductance and intracellular free Ca2+in outer hair cells of the guinea pig cochlea

Abstract

We have studied the action of cholinergic agonists on outer hair cells, both in situ and isolated from the cochlea of the guinea pig, combining new fast CCD technology for Ca2+imaging and conventional patch-clamp methods. Carbachol (1mM) activated a current with a reversal potential near –70mV and a bell-shaped I-V curve, suggesting that it was a Ca2+-activated K+current. In a few cells, this current was preceded by a transient inward current, probably owing to an influx of Ca2+and other cations through the acetylcholine (ACh) receptors. The amplitude of the Ca2+signal was maximal in a circumscribed region at the basal pole of the cell and decreased steeply towards the apical pole, compatible with Ca2+-influx and/or Ca2+-induced Ca2+-release at the cells base. The time course of the Ca2+-rise was fastest at the base, but it was still slightly slower, and more rounded, than that of the K+current. In some recordings the K+current was observed without any measurable change of intracellular Ca2+. The K+current was potentiated (18%) by caffeine (5mM), and decreased (19%) by ryanodine (0.1mM) in the majority of cells tested. The results are discussed in terms of a labile intracellular Ca2+store located at the base of the cell, close to the Ca2+-permeable ACh receptor channels and Ca2+-activated K+channels, whose contribution to the Ca2+rise occurring in the region of the channels is variable, and probably dependent on its ability to refill with Ca2+.