Abstract
The patch clamp technique introducted by Neher and colleagues (Neher, Sakmann and Steinbach, 1978; Hamill et al., 1981) has been used to study ion channels at both the single-channel and the whole-cell level in animal, plant and bacterial cells. The successful application of the technique is critically dependent on the ability to form a high resistance (10–100 GΩ) seal between the patch pipette and the plasma membrane of the cell under study. The formation of a giga-seal effectively isolates the membrane patch both electrically and chemically. Electrical isolation of the patch allows the current flowing through a single channel to be resolved and the patch to be voltage clamped by simply applying a voltage to the pipette. Chemical isolation of a patch of membrane from a cell allows the normal ionic environment of the patch to be manipulated. The giga-seal is also mechanically very stable and enables the patch to be either excised from the cell or ruptured, thus creating a number of different and useful recording configurations.
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References
Auerbach, A. and Sachs, F. (1984) Patch clamp studies of single ionic channels. Ann. Rev. Bioeng. 13:269–302.
Colquhoun, D. and Hawkes, A.G. (1983) The principles of the stochastic interpretation of ion-channel mechanisms, in Single-Channel Recording, (eds B. Sakmann and E. Neher), Plenum Press, New York, pp. 135–76.
Colquhoun, D. and Sigworth, F.J. (1983) Fitting and statistical analysis of single channel records, in Single-Channel Recording, (eds B. Sakmann and E. Neher), Plenum Press, New York, pp. 191–263.
Copello, J., Simon, B., Segal,Y. et al .(1991) Ba2+ release from soda glass modifies single maxi K+ channel activity in patch clamp experiments. Biophys. J. 60:931–41.
Copello, J., Wehner, F. and Reuss, L. (1993) Artifactual expression of a maxi-K+ channels in basolateral membrane of gallbladder epithelia cells. Am. J. Physiol. 264:C1128-C1136.
Corey, D.P. and Stevens, C.F. (1983) Science and technology of patch recording electrodes, in Single-Channel Recording, (eds B. Sakmann and E. Nener), Plenum Press, New York, pp. 53–68.
Cota, G. and Armstrong, CM. (1987) Potassium channel ‘inactivation’ induced by soft glass patch pipettes. Biophys. J. 53:107–9.
Dionne, V.E. (1981) The kinetics of slow muscle acetylcholine-operated channels in the garter snake. J. Physiol. 310:159–90.
Donaldson, P.J. and Lewis, S.A. (1990) The effect of serosal hypertonic challenge on basolateral membrane potential in the rabbit urinary bladder. Am. J. Physiol. 258: C248-C257.
Dragsten, P.R., Blumenthal, R. and Handler, J.S. (1981) Membrane asymmetry in epithelia: is the tight junction a barrier to diffusion in the plasma membrane. Nature 294:718–22.
Dunne, M.J. and Peterson, O.H. (1986) GTP and GDP activation of K+ that can be inhibited by ATP. Pflügers Arch. 407: 564–5.
Fenwick, E.M., Marty, A. and Neher, E. (1982) A patch-clamp study of bovine chromaffin cells and their sensitivity to acetylcholine. J. Physiol (Lond.) 331:577–97.
Gogelein, H. and Greger, R. (1984) Single channel recordings from basolateral and apical membrane of renal proximal tubules. Pflüger Arch. 408:282–90.
Goldman, D.E. (1943) Potential, impedance, and rectification in membranes. J. Gen. Physiol. 27:37–60.
Hamill, O.P. (1983) Potassium and chloride channels in red blood cells, in Single-channel Recording (eds B. Sakmann and E. Neher), Plenum Press, New York, pp. 451–71.
Hamill, O.P., Marty, A., Neher, E. et al .(1981) Improved patch clamp techniques for high resolution current recording from cells and cell-free membrane patches. Pflügers Arch. 391:85–100.
Horn, R. and Marty, A. (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J. Gen. Physiol. 92:145–59.
Hunter, M., Lopes, A.G., Boulpaep, E.L. and Giebisch, G.H. (1984) Single channel recordings of calcium-activated potassium channels in the apical membrane of rabbit cortical collecting tubules. PNAS 81:4237–9.
Kakei, M. and Ashcroft, F.M. (1987) A microflow superfusion system for use with excised membrane patches. Pflügers Arch. 409:337–41.
Labarca, P., Coronado, R. and Miller, C. (1980) Thermodynamic and kinetic studies of the gating behavior of a K+-selective channel from the sacroplasmic reticulum membrane. J. Gen. Physiol. 76:397–424.
Lapointe, J.-Y. and Szabo, G. (1987) A novel holder for allowing internal perfusion of patch pipettes. Pflügers Arch. 410:212–16.
Lewis, S.A. and Alles, W.P. (1986) Urinary kallikrein: a physiological regulator of epithelial Na+ transport. PNAS 83:5345–8.
Lindau, M. and Fernandez, J.M. (1986) IgE-mediated degranulation of mast cells does not require opening of ion channels. Nature 319:150–3.
Lindemann, B. and Van Driessche, W. (1977) Sodium-specific membrane channels of frog skin are pores: current fluctuations reveal high turnover. Science 221:292–4.
Lopes, A.G. and Guggino, W.B. (1987) Volume regulation in the early proximal tubule of the Necturus kidney. J. Memb. Biol. 97:117–25.
Neher, E., Sakmann, B. and Steinbach, J.H. (1978) The extracellular patch clamp: a method for resolving currents through individual open channels in biological membranes. Pflügers Arch. 375:219–26.
Palmer, L.G. and Frindt, G. (1986) Amiloride-sensitive Na+ channels from the apical membrane of the rat cortical collecting tubule. PNAS 83:2767–70.
Rae, J.L. (1985) The application of patch clamp methods to ocular epithelia. Curr. Eye Res. 4:409–20.
Rae, J.L. and Levis, R.A. (1984) Patch clamp recordings from the epithelium of the lens obtained using glasses selected for low noise and improved sealing properties. Biophys. J. 45:144–6.
Richards, N.W. and Dawson, D.C. (1986) Single potassium channels blocked by lidocaine and quinidine in isolated turtle colon epithelial cells. Am. J. Physiol. 251:C85-C89.
Sackin, H. and Palmer, L.G. (1987) Basolateral potassium channels in renal proximal tubules. Am. J. Physiol. 253:F476-F487.
Sakmann, B. and Neher, E. (1983) Geometric parameters of pipettes and membrane patches, in Single-Channel Recording, (eds B. Sakmann and E. Neher), Plenum Press, New York, pp. 37–52.
Sakmann, B. and Neher, E. (1984) Patch clamp techniques for studying ionic channels in excitable membranes. Ann. Rev. Physiol. 46:455–72.
Sigworth, F.J. and Neher, E. (1980) Single Na+ channel currents observed in cultured rat muscle cells. Nature 287:447–9.
Trautmann, A. and Siegelbaum, S.A. (1983) The influence of membrane patch isolation on single acetylcholine-channel current in rat myotubules, in Single-channel Recording (eds B. Sakmann and E. Neher), Plenum Press, New York, pp. 473–80.
Yellen, G. (1982) Single Ca2+ activated non-selective cation channels in neuroblastoma. Nature 296:357–9.
Yellen, G. (1984) Ionic permeation and blockade in Ca2+-activated K+ channels of Bovine chromaffin cells. J. Gen. Physiol. 84:157–86.
Ziomek, C.A., Schulman, S. and Edidin, M. (1980). Redistribution of membrane proteins in isolated mouse intestinal epithelial cells. J. Cell Biol. 86:849–57.
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Lewis, S.A. (1996). Methods and experimental analysis of single ion channels. In: Wills, N.K., Reuss, L., Lewis, S.A. (eds) Epithelial Transport. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-1495-7_10
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DOI: https://doi.org/10.1007/978-94-009-1495-7_10
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-7168-0
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