Abstract
1. Effects of amiloride, applied extracellularly, on mechano-electrical transduction (MET) currents were investigated in dissociated hair cells of a chick with a whole-cell patch-electrode voltage clamp technique. Amiloride blocked the MET channel. The blocking was reversible and was both dose and voltage dependent and specific to the MET channel. The voltage-dependent Ca2+ channel of the basolateral membrane was not affected within the concentration range studied (up to 0.7 mM). 2. The limiting conductance of the MET at large negative membrane potentials decreased with increasing amiloride concentration. A dose-response relationship of the relative MET conductance (defined as the ratio of the MET channel conductance in the presence of amiloride to that without) at membrane potentials more negative than -50 mV had a Hill coefficient of 1, and a dissociation constant (KD) of 5 x 10(-5) M. 3. When amiloride was applied, the MET conductance increased as the membrane was depolarized, and the limiting value at positive membrane potentials was close to that of the control. The relationship between the relative MET conductance and the membrane potential was S-shaped. The conductance vs. voltage relationship was shifted in a positive direction along the voltage axis as the amiloride concentration was increased. 4. The blocking effect of amiloride on the MET channel was apparently independent of the mechanical gating of the channel. The voltage-independent block at or near the resting membrane potential and a voltage-dependent lifting of the block at depolarized membrane potentials could be explained quantitatively by a kinetic model which postulates one blocked state and two open states which have different amiloride affinities.
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Selected References
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- Art J. J., Fettiplace R. Variation of membrane properties in hair cells isolated from the turtle cochlea. J Physiol. 1987 Apr;385:207–242. doi: 10.1113/jphysiol.1987.sp016492. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benos D. J. Amiloride: a molecular probe of sodium transport in tissues and cells. Am J Physiol. 1982 Mar;242(3):C131–C145. doi: 10.1152/ajpcell.1982.242.3.C131. [DOI] [PubMed] [Google Scholar]
- Bezanilla F., Armstrong C. M. Negative conductance caused by entry of sodium and cesium ions into the potassium channels of squid axons. J Gen Physiol. 1972 Nov;60(5):588–608. doi: 10.1085/jgp.60.5.588. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CITRON L., EXLEY D., HALLPIKE C. S. Formation, circulation and chemical properties of the labyrinthine fluids. Br Med Bull. 1956 May;12(2):101–104. doi: 10.1093/oxfordjournals.bmb.a069529. [DOI] [PubMed] [Google Scholar]
- Corey D. P., Hudspeth A. J. Ionic basis of the receptor potential in a vertebrate hair cell. Nature. 1979 Oct 25;281(5733):675–677. doi: 10.1038/281675a0. [DOI] [PubMed] [Google Scholar]
- DeLong J., Civan M. M. Apical sodium entry in split frog skin: current-voltage relationship. J Membr Biol. 1984;82(1):25–40. doi: 10.1007/BF01870729. [DOI] [PubMed] [Google Scholar]
- Hagiwara S., Ohmori H. Studies of calcium channels in rat clonal pituitary cells with patch electrode voltage clamp. J Physiol. 1982 Oct;331:231–252. doi: 10.1113/jphysiol.1982.sp014371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hamilton K. L., Eaton D. C. Single-channel recordings from amiloride-sensitive epithelial sodium channel. Am J Physiol. 1985 Sep;249(3 Pt 1):C200–C207. doi: 10.1152/ajpcell.1985.249.3.C200. [DOI] [PubMed] [Google Scholar]
- Hudspeth A. J., Corey D. P. 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):2407–2411. doi: 10.1073/pnas.74.6.2407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hudspeth A. J. Extracellular current flow and the site of transduction by vertebrate hair cells. J Neurosci. 1982 Jan;2(1):1–10. doi: 10.1523/JNEUROSCI.02-01-00001.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lewis R. S., Hudspeth A. J. Voltage- and ion-dependent conductances in solitary vertebrate hair cells. Nature. 1983 Aug 11;304(5926):538–541. doi: 10.1038/304538a0. [DOI] [PubMed] [Google Scholar]
- Lindemann B., Van Driessche W. Sodium-specific membrane channels of frog skin are pores: current fluctuations reveal high turnover. Science. 1977 Jan 21;195(4275):292–294. doi: 10.1126/science.299785. [DOI] [PubMed] [Google Scholar]
- Ohmori H. Gating properties of the mechano-electrical transducer channel in the dissociated vestibular hair cell of the chick. J Physiol. 1987 Jun;387:589–609. doi: 10.1113/jphysiol.1987.sp016590. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohmori H. Mechanical stimulation and Fura-2 fluorescence in the hair bundle of dissociated hair cells of the chick. J Physiol. 1988 May;399:115–137. doi: 10.1113/jphysiol.1988.sp017071. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohmori H. Mechano-electrical transduction currents in isolated vestibular hair cells of the chick. J Physiol. 1985 Feb;359:189–217. doi: 10.1113/jphysiol.1985.sp015581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohmori H. Studies of ionic currents in the isolated vestibular hair cell of the chick. J Physiol. 1984 May;350:561–581. doi: 10.1113/jphysiol.1984.sp015218. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palmer L. G. Ion selectivity of the apical membrane Na channel in the toad urinary bladder. J Membr Biol. 1982;67(2):91–98. doi: 10.1007/BF01868651. [DOI] [PubMed] [Google Scholar]
- Palmer L. G. Voltage-dependent block by amiloride and other monovalent cations of apical Na channels in the toad urinary bladder. J Membr Biol. 1984;80(2):153–165. doi: 10.1007/BF01868771. [DOI] [PubMed] [Google Scholar]
- Sariban-Sohraby S., Benos D. J. The amiloride-sensitive sodium channel. Am J Physiol. 1986 Feb;250(2 Pt 1):C175–C190. doi: 10.1152/ajpcell.1986.250.2.C175. [DOI] [PubMed] [Google Scholar]
- Warncke J., Lindemann B. Voltage dependence of Na channel blockage by amiloride: relaxation effects in admittance spectra. J Membr Biol. 1985;86(3):255–265. doi: 10.1007/BF01870605. [DOI] [PubMed] [Google Scholar]
- Woodhull A. M. Ionic blockage of sodium channels in nerve. J Gen Physiol. 1973 Jun;61(6):687–708. doi: 10.1085/jgp.61.6.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
