Release and elementary mechanisms of nitric oxide in hair cells.
The enzyme nitric oxide (NO) synthase, that produces the signaling molecule NO, has been identified in several cell types in the inner ear. However, it is unclear whether a measurable quantity of NO is released in the inner ear to confer specific functions. Indeed, the functional significance of NO and the elementary cellular mechanism thereof are most uncertain. Here, we demonstrate that the sensory epithelia of the frog saccule release NO and explore its release mechanisms by using self-referencing NO-selective electrodes. Additionally, we investigated the functional effects of NO on electrical properties of hair cells and determined their underlying cellular mechanism. We show detectable amounts of NO are released by hair cells (>50 nM). Furthermore, a hair-cell efferent modulator acetylcholine produces at least a threefold increase in NO release. NO not only attenuated the baseline membrane oscillations but it also increased the magnitude of current required to generate the characteristic membrane potential oscillations. This resulted in a rightward shift in the frequency-current relationship and altered the excitability of hair cells. Our data suggest that these effects ensue because NO reduces whole cell Ca(2+) current and drastically decreases the open probability of single-channel events of the L-type and non L-type Ca(2+) channels in hair cells, an effect that is mediated through direct nitrosylation of the channel and activation of protein kinase G. Finally, NO increases the magnitude of Ca(2+)-activated K(+) currents via direct NO nitrosylation. We conclude that NO-mediated inhibition serves as a component of efferent nerve modulation of hair cells.
Lv
Ping
P
Center for Neuroscience, Department of Anesthesiology and Pain Medicine, Program in Communication Sciences, University of California, Davis, Davis, CA 95616, USA.
Rodriguez-Contreras
Adrian
A
Kim
Hyo Jeong
HJ
Zhu
Jun
J
Wei
Dongguang
D
Choong-Ryoul
Sihn
S
Eastwood
Emily
E
Mu
Karen
K
Levic
Snezana
S
Song
Haitao
H
Yevgeniy
Petrov Y
PY
Smith
Peter J S
PJ
Yamoah
Ebenezer N
EN
D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D004566:Electrodes; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D018069:Hair Cells, Vestibular / Q000187:drug effects / Q000502:physiology*; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009569:Nitric Oxide / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010507:Periodicity; D011188:Potassium / Q000378:metabolism; D011336:Probability; D011892:Rana catesbeiana; D012444:Saccule and Utricle / Q000187:drug effects / Q000502:physiology
D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D009569:Nitric Oxide; D017869:Cyclic GMP-Dependent Protein Kinases; D000109:Acetylcholine; D011188:Potassium; D002118:Calcium