Abstract
Receptor currents in response to mechanical stimuli were recorded from hair cells in the excised epithelium of the bull-frog sacculus by the whole-cell, gigohm-seal voltage-clamp technique. The stimulus-dependent transduction current was separated from the cell's stimulus-independent K+ and Ca2+ currents; the K+ currents were blocked with an internal solution containing Cs+ while the Ca2+ current was reduced by holding the membrane potential below -70 mV. The temperature of the preparation was maintained at about 10 degrees C to slow the kinetics of the cells' transduction channels. Calibrated displacements of hair bundles of individual hair cells were made with a probe coupled by suction to the kinociliary bulb and moved with a piezoelectricbimorph stimulator. The root mean square noise of probe motion was less than 2 nm. The mean, I, and the variance, sigma 2, of the receptor current were measured from the response to saturating (+/- 0.5 micron) displacements of the hair bundle. I was corrected for current offsets and sigma 2 for the transduction-independent background variance. The relation between sigma 2 and I is consistent with the predictions of a two-conductance-state model of the transduction channel, a model having only one non-zero conductance state. The relation between sigma 2 and I was fitted by the equation sigma 2 = Ii-I2/N, where N is the number of transduction channels in the cell and i is the current through a single open channel. The conductance of the transduction channel is approximately ohmic with a reversal potential near 0 mV. The estimated conductance of a single transduction channel, gamma, is 12.7 +/- 2.7 pS (mean +/- S.D.; n = 18) at 10 degrees C. gamma is independent of the maximum transduction conductance of the cell, Gmax. The number of transduction channels, N, is proportional to Gmax. N ranges from 7 to 280 in cells with Gmax ranging from 0.08 to 2.48 nS. The largest values of N correspond to a few, perhaps four, active transduction channels per stereocilium. Control experiments show that transduction by the hair cell of two artifactual sources of hair-bundle stimulation, noisy or discontinuous motion of the probe, do not contribute substantially to the measured variance, sigma 2. Displacement-response curves are generally sigmoidal and symmetrical; they reasonably fit the predictions of a two-kinetic-state model, comprising one open state and one closed state. The estimated displacement-sensitive free energy, Z, is 5.7 +/- 1.1 kcal/mol micron (mean +/- S.D., n = 18).(ABSTRACT TRUNCATED AT 400 WORDS)
MeSH Terms
Action Potentials
Animals
Electricity
Hair Cells, Auditory/physiology
In Vitro Techniques
Ion Channels/physiology
Kinetics
Models, Neurological
Physical Stimulation
Rana catesbeiana
Saccule and Utricle/physiology
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Holton T
Hudspeth A J
References (24)
24 references, click to expand
-
Experiments on ototoxic effects of antibiotics.
Adv Otorhinolaryngol. 1973;20:14-41
PMID: 4575758
-
Mechano-electrical transduction currents in isolated vestibular hair cells of the chick.
J Physiol. 1985 Feb;359:189-217
PMID: 2582113
-
Conductance fluctuations and ionic pores in membranes.
Annu Rev Biophys Bioeng. 1977;6:345-81
PMID: 68708
-
Sensitivity, polarity, and conductance change in the response of vertebrate hair cells to controlled mechanical stimuli.
Proc Natl Acad Sci U S A. 1977 Jun;74(6):2407-11
PMID: 329282
-
Relaxation and fluctuations of membrane currents that flow through drug-operated channels.
Proc R Soc Lond B Biol Sci. 1977 Nov 14;199(1135):231-62
PMID: 22856
-
Stereocilia mediate transduction in vertebrate hair cells (auditory system/cilium/vestibular system).
Proc Natl Acad Sci U S A. 1979 Mar;76(3):1506-9
PMID: 312502
-
Reversal of hair cell responses by current [proceedings].
J Physiol. 1979 Oct;295:66P
PMID: 521982
-
Ionic basis of the receptor potential in a vertebrate hair cell.
Nature. 1979 Oct 25;281(5733):675-7
PMID: 45121
-
The variance of sodium current fluctuations at the node of Ranvier.
J Physiol. 1980 Oct;307:97-129
PMID: 6259340
-
Mechanical stimulation and micromanipulation with piezoelectric bimorph elements.
J Neurosci Methods. 1980 Dec;3(2):183-202
PMID: 7206783
-
Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
Pflugers Arch. 1981 Aug;391(2):85-100
PMID: 6270629
-
Non-linearities in the responses of turtle hair cells.
J Physiol. 1981 Jun;315:317-38
PMID: 7310712
-
Extracellular current flow and the site of transduction by vertebrate hair cells.
J Neurosci. 1982 Jan;2(1):1-10
PMID: 6275046
-
Directional sensitivity of individual vertebrate hair cells to controlled deflection of their hair bundles.
Ann N Y Acad Sci. 1981;374:1-10
PMID: 6978627
-
A patch-clamp study of bovine chromaffin cells and of their sensitivity to acetylcholine.
J Physiol. 1982 Oct;331:577-97
PMID: 6296371
-
Origin of the receptor potential in inner hair cells of the mammalian cochlea--evidence for Davis' theory.
Nature. 1983 Jan 27;301(5898):334-6
PMID: 6823305
-
Mechanoelectrical transduction by hair cells in the acousticolateralis sensory system.
Annu Rev Neurosci. 1983;6:187-215
PMID: 6301349
-
Analysis of the microphonic potential of the bullfrog's sacculus.
J Neurosci. 1983 May;3(5):942-61
PMID: 6601693
-
Kinetics of the receptor current in bullfrog saccular hair cells.
J Neurosci. 1983 May;3(5):962-76
PMID: 6601694
-
Voltage- and ion-dependent conductances in solitary vertebrate hair cells.
Nature. 1983 Aug 11-17;304(5926):538-41
PMID: 6603579
-
Mechanoelectrical transducer has discrete conductances in the chick vestibular hair cell.
Proc Natl Acad Sci U S A. 1984 Mar;81(6):1888-91
PMID: 6584923
-
Stretch-activated single ion channel currents in tissue-cultured embryonic chick skeletal muscle.
J Physiol. 1984 Jul;352:685-701
PMID: 6086918
-
Cross-links between stereocilia in the guinea pig organ of Corti, and their possible relation to sensory transduction.
Hear Res. 1984 Aug;15(2):103-12
PMID: 6436216
-
Regional distribution of putrescine, spermidine and spermine in relation to the distribution of RNA and DNA in the rat nervous system.
J Neurochem. 1975 Apr;24(4):791-5
PMID: 1123635