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Journal of Neuroscience, Vol 16, 2574-2584, Copyright © 1996 by Society for Neuroscience
Acetylcholine-induced potassium current of guinea pig outer hair cells: its dependence on a calcium influx through nicotinic-like receptors
C Blanchet, C Erostegui, M Sugasawa and D Dulon
Laboratoire d'Audiologie Experimentale, Institute National de la Sante et de la Recherche Medicale, Bordeaux, France.
The cholinergic efferent inhibition of mammalian outer hair cells (OHCs) is
mediated by a hyperpolarizing K+ current. We have made whole- cell
tight-seal recordings from single OHCs isolated from the guinea pig cochlea
to characterize the mechanism by which acetylcholine (ACh) activates K+
channels. After ACh application, OHCs exhibited a biphasic response: an
early depolarizing current preceding the predominant hyperpolarizing K+
current. The current-voltage (I-V) relationship of the ACh-induced response
displayed an N-shape, suggesting the involvement of Ca2+ influx. When
whole-cell recording was combined with confocal calcium imaging, we
simultaneously observed the ACh-induced K+ current (IK(ACh)) and a Ca2+
response restricted to the synaptic area of the cell. This IK(ACh) could be
prevented by loading OHCs with 10 mM of the fast Ca2+ buffer
bis(2-aminophenoxy)ethane-N,N,N',N'-tetra- acetic acid (or BAPTA),
therefore allowing the observation of the ACh- induced early current in
isolation. This early current revealed nicotinic features because it
activated with an intrinsic delay in the millisecond range, reversed nearly
in between potassium and sodium equilibrium potentials, and was blocked by
curare. However, it was strongly reduced in the absence of external Ca2+,
and its I-V relationship displayed an unusual outward rectification at
positive membrane potentials and an inward rectification below -60 mV. The
results indicate that the cholinergic response of mammalian OHCs involves a
"nicotinic-like" nonspecific cation channel through which Ca2+ enters and
triggers activation of nearby Ca2+-dependent K+ channels.
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