The Journal of Neuroscience, April 15, 2001, 21(8):2640-2650
Contribution of the Plasmalemma to Ca2+ Homeostasis
in Hair Cells
Catherine
Boyer1, 2,
Jonathan J.
Art2,
Claude J.
Dechesne1,
Jacques
Lehouelleur1,
Jean
Vautrin1, and
Alain
Sans1
1 Institut National de la Santé et de la
Recherche Médicale U-432, Université Montpellier II, 34095 Montpellier cedex 05, France, and 2 University of Illinois,
College of Medicine, Department of Anatomy and Cell Biology,
Chicago, Illinois 60612
Calcium influx through transduction channels and efflux via
plasmalemmal Ca2+-ATPases (PMCAs) are known to
contribute to calcium homeostasis and modulate sensory transduction in
vertebrate hair cells. To examine the relative contributions of apical
and basolateral pathways, we analyzed the calcium dynamics in solitary
ciliated and deciliated guinea pig type I and type II vestibular hair
cells. Whole-cell patch-clamp recordings demonstrated that these cells
had resting potentials near
70 mV and could be depolarized by 10-20
mV by superfusion with high potassium. Fura-2 measurements indicated that ciliated type II cells and deciliated cells of either type had low
basal [Ca2+]i, near ~90
nM, and superfusion with high potassium led to transient calcium increases that were diminished in the presence of
Ca2+ channel blockers. In contrast, measurements of
type I ciliated cells, hair cells with large calyceal afferents, were
associated with a higher basal [Ca2+]i
of ~170 nM. High-potassium superfusion of these cells
induced a paradoxical decrease in
[Ca2+]i that was augmented in the
presence of Ca2+ channel blockers. Optical
localization of dihydropyridine binding to the kinocilium suggests that
they contain L-type calcium channels, and as a result apical calcium
influx includes a contribution from voltage-dependent ion channels in
addition to entry via transduction channels localized to the
stereocilia. Eosin block of PMCA significantly altered both
[Ca2+]i baseline and transient
responses only in ciliated cells suggesting that, in agreement with
immunohistochemical studies, PMCA is primarily localized to the bundles.
Key words:
PMCA; calcium channels; vestibular hair cells; bundles; fura-2 fluorescence; guinea pig
Copyright © 2001 Society for Neuroscience 0270-6474/01/2182640-11$05.00/0