Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1991 Sep;60(3):660–670. doi: 10.1016/S0006-3495(91)82095-1

Single-channel dose-response studies in single, cell-attached patches.

A Auerbach 1
PMCID: PMC1260109  PMID: 1718468

Abstract

A method for carrying out dose-response studies of ion channel currents in cell-attached patches has been devised. Patch pipettes are filled at the tip with a solution containing one concentration of ligand and then backfilled with another. The concentration of ligand at the membrane is described as a function of time by the equation for diffusion in a cone, allowing response vs. time data to be transformed into a dose-response curve. For Xenopus myocyte cholinergic receptors, examples of the use of this method are given for several concentration-dependent reactions including blockade by the local anesthetic QX-222, activation by acetylcholine, and modulation of current amplitude by sodium ions. Several methods of analyzing the nonstationary channel kinetics are presented, including a pseudo-stationary approach that uses interval likelihood maximization.

Full text

PDF
670

Images in this article

662FIGURE 1
on p.662

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Auerbach A., Lingle C. J. Activation of the primary kinetic modes of large- and small-conductance cholinergic ion channels in Xenopus myocytes. J Physiol. 1987 Dec;393:437–466. doi: 10.1113/jphysiol.1987.sp016832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auerbach A., Lingle C. J. Heterogeneous kinetic properties of acetylcholine receptor channels in Xenopus myocytes. J Physiol. 1986 Sep;378:119–140. doi: 10.1113/jphysiol.1986.sp016211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chay T. R. Kinetic modeling for the channel gating process from single channel patch clamp data. J Theor Biol. 1988 Jun 22;132(4):449–468. doi: 10.1016/s0022-5193(88)80084-5. [DOI] [PubMed] [Google Scholar]
  4. Covarrubias M., Steinbach J. H. Excision of membrane patches reduces the mean open time of nicotinic acetylcholine receptors. Pflugers Arch. 1990 Jun;416(4):385–392. doi: 10.1007/BF00370744. [DOI] [PubMed] [Google Scholar]
  5. Cull-Candy S. G., Miledi R., Parker I. Single glutamate-activated channels recorded from locust muscle fibres with perfused patch-clamp electrodes. J Physiol. 1981 Dec;321:195–210. doi: 10.1113/jphysiol.1981.sp013979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dionne V. E. Characterization of drug iontophoresis with a fast microassay technique. Biophys J. 1976 Jul;16(7):705–717. doi: 10.1016/S0006-3495(76)85723-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horn R., Lange K. Estimating kinetic constants from single channel data. Biophys J. 1983 Aug;43(2):207–223. doi: 10.1016/S0006-3495(83)84341-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Horn R., Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988 Aug;92(2):145–159. doi: 10.1085/jgp.92.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Horn R., Patlak J. Single channel currents from excised patches of muscle membrane. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6930–6934. doi: 10.1073/pnas.77.11.6930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horn R., Vandenberg C. A. Statistical properties of single sodium channels. J Gen Physiol. 1984 Oct;84(4):505–534. doi: 10.1085/jgp.84.4.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liu Y., Dilger J. P. Opening rate of acetylcholine receptor channels. Biophys J. 1991 Aug;60(2):424–432. doi: 10.1016/S0006-3495(91)82068-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mathias R. T., Cohen I. S., Oliva C. Limitations of the whole cell patch clamp technique in the control of intracellular concentrations. Biophys J. 1990 Sep;58(3):759–770. doi: 10.1016/S0006-3495(90)82418-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Neher E., Steinbach J. H. Local anaesthetics transiently block currents through single acetylcholine-receptor channels. J Physiol. 1978 Apr;277:153–176. doi: 10.1113/jphysiol.1978.sp012267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pusch M., Neher E. Rates of diffusional exchange between small cells and a measuring patch pipette. Pflugers Arch. 1988 Feb;411(2):204–211. doi: 10.1007/BF00582316. [DOI] [PubMed] [Google Scholar]
  15. Roux B., Sauvé R. A general solution to the time interval omission problem applied to single channel analysis. Biophys J. 1985 Jul;48(1):149–158. doi: 10.1016/S0006-3495(85)83768-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ruknudin A., Song M. J., Sachs F. The ultrastructure of patch-clamped membranes: a study using high voltage electron microscopy. J Cell Biol. 1991 Jan;112(1):125–134. doi: 10.1083/jcb.112.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sachs F., Neil J., Barkakati N. The automated analysis of data from single ionic channels. Pflugers Arch. 1982 Dec;395(4):331–340. doi: 10.1007/BF00580798. [DOI] [PubMed] [Google Scholar]
  18. Sakmann B., Patlak J., Neher E. Single acetylcholine-activated channels show burst-kinetics in presence of desensitizing concentrations of agonist. Nature. 1980 Jul 3;286(5768):71–73. doi: 10.1038/286071a0. [DOI] [PubMed] [Google Scholar]
  19. Tang J. M., Wang J., Quandt F. N., Eisenberg R. S. Perfusing pipettes. Pflugers Arch. 1990 May;416(3):347–350. doi: 10.1007/BF00392072. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES