Summary
The behavior of single Cl− channel was studied by fusing isolated canine cardiac sarcoplasmic reticulum (SR) vesicles into planar lipid bilayers. The channel exhibited unitary conductance of 55 pS (in 260mm Cl−) and steady-state activation. Subconductance states were observed. Open probability was dependent on holding potentials (−60 to +60 mV) and displayed a bell-shaped relationship, with probability values ranging from 0.2 to 0.8 with a maximum at −10 mV. Channel activity was irreversibly inhibited by DIDS, a stilbene derivative. Time analysis revealed the presence of one time constant for the full open state and three time constants for the closed states. The open and the longer closed time constants were found to be voltage dependent. The behavior of the channel was not affected by changing Ca2+ and Mg2+ concentrations in both chambers, nor by adding millimolar adenosine triphosphate, or by changing the pH from 7.4 to 6.8. The presence of sulfate anions decreased the unit current amplitude, but did not affect the open probability. These results reveal that at the unitary level the cardiac SR anion-selective channel has distinctive as well as similar electrical properties characteristic of other types of Cl− channels.
Similar content being viewed by others
References
Barrett, J. N., Magleby, K. L., Palotta, B. S. 1982. Properties of single calcium-activated potassium channels in cultured rat muscle.J. Physiol. (London) 331:211–230
Bezanilla, F. 1985. A high capacity data recording device based on a digital audio processor and a video cassette recorder.Biophys. J. 47:437–441
Blatz, A. L., Magleby, K. L. 1986. Quantitative description of three modes of activity of fast chloride channels from rat skeletal muscle.J. Physiol. (London) 378:141–174
Bormann, J., Hamill, O. P., Sakmann, B. 1987. Mechanism of anion permeation through channels gated by glycine and α-aminobutyric acid in mouse cultured spinal neurones.J. Physiol. (London) 385:243–286
Bretag, A. M. 1987. Muscle chloride channels.Physiol. Rev. 67:618–723
Caillé, J. P. 1985. Intracellular chloride activity in rabbit papillary muscle: Effect of serum.Can. J. Physiol. Pharmacol. 64:1381–1384
Colombini, M. 1979. A candidate for the permeability pathway of the outer mitochondrial membrane.Nature (London) 279:643–645
Coronado, R., Latorre, R. 1982. Detection of K+ and Cl− channels from calf cardiac sarcolemma in planar lipid bilayer membranes.Nature (London) 298:849–851
Coronado, R., Miller, C. 1982. Conduction and block by organic cations in a K+ selective channel from sarcoplasmic reticulum.J. Gen. Physiol. 79:529–547
Coronado, R., Rosenberg, R., Miller, C. 1980. Ionic selectivity, saturation and block in a K+ selective channel from sarcoplasmic reticulum.J. Gen. Physiol. 76:425–446
Coulombe, A., Duclohier, H., Coraboeuf, E., Touzet, N. 1987. Single Cl− permeable channels of large conductance in cultured cardiac cells of new-born rats.Eur. Biophys. J. 14:155–162
Endo, M. 1977. Calcium release from the SR.Physiol. Rev. 57:71–108
Fabiato, A. 1985. Time and calcium dependence of activation and inactivation of Ca2+ induced Ca2+ release of Ca from the SR of a skinned canine cardiac purkinje cell.J. Gen. Physiol. 85:247–289
Garcia, A. M., Miller, C. 1984. Channel mediated monovalent cation fluxes in isolated sarcoplasmic reticulum vesicles.J. Gen. Physiol. 83:819–839
Hals, G. D., Palade, P. T. 1988. Novel Cl− channel from skeletal muscle SR recorded in native membrane.Biophys. J. 53:338a
Kasai, M., Kanemasa, T., Fukumoto, S. 1979. Determination of reflection coefficients for various ions and neutral molecules in sarcoplasmic reticulum vesicles through osmotic volume change studies by stopped flow technique.J. Membrane Biol. 51:311–324
Kasai, M., Kometani, T. 1979. Inhibition of anion permeability of sarcoplasmic reticulum vesicles by 4-acetoamido-4-isothiocyanostilbene-2,2-disulfonate.Biochim. Biophys. Acta 557:243–247
Kasai, M., Tagushi, T. 1981. Inhibition of anion permeability of sarcoplasmic reticulum vesicles by stilbene derivatives and the identification of an inhibitor-binding protein.Biochim. Biophys. Acta 643:213–219
Knauf, P. A., Rothstein, A. 1971. Chemical modification of membranes I. Effects of sulphydryl and aminoreactive reagents on anion and cation permeability of the human red blood cells.J. Gen. Physiol. 58:190–210
Kometani, T., Kasai, M. 1978. Ionic permeability of sarcoplasmic reticulum vesicles measured by light scattering method.J. Membrane Biol. 41:295–308
Lai, F. A., Anderson, K., Rousseau, E., Liu, Q. Y., Meissner, G. 1988. Evidence for a Ca2+ channel within the ryanodine receptor complex from cardiac sarcoplasmic reticulum.Biochem. Biophys. Res. Commun. 151:441–449
Magleby, K. L., Pallota, B. S. 1983. Calcium dependence of open and shut interval distribution from calcium activated potassium channels in cultured rat muscle.J. Physiol. (London) 344:585–604
McKinley, D., Meissner, G. 1978. Evidence for a K+, Na+ permeable channel in sarcoplasmic reticulum.J. Membrane Biol. 44:159–186
Meissner, G. 1983. Monovalent ion and calcium ion fluxes in sarcoplasmic reticulum.Mol. Cell. Biochem. 55:65–82
Meissner, G., Henderson, J. 1987. Rapid calcium release from cardiac sarcoplasmic reticulum vesicles is dependent on Ca2+ and is modulated by Mg2+, adenine nucleotide and calmodulin.J. Biol. Chem. 262:3065–3073
Meissner, G., McKinley, D. 1982. Permeability of canine cardiac sarcoplasmic reticulum vesicles to K+, Na+, H+ and Cl−.J. Biol. Chem. 257:7704–7711
Miller, C. 1978. Voltage gated cation conductance channel from fragmented sarcoplasmic reticulum: Steady state electrical properties.J. Membrane Biol. 40:1–23
Miller, C., Raker, E. 1976. Ca2+ induced fusion of fragmented SR with artificial bilayer.J. Membrane Biol. 30:283–300
Miller, C., White, M. W. 1984. Dimeric structure of single chloride channels fromTorpedo electroplax.Proc. Natl. Acad. Sci. USA 81:2772–2775
Nelson, D. J., Tang, J. M., Palmer, L. G. 1984. Single channel recordings of apical membrane chloride conductance in A6 epithelial cells.J. Membrane Biol. 80:81–89
Rousseau, E., Roberson, M., Meissner, G. 1988. Properties of single chloride selective channel from sarcoplasmic reticulum.Eur. Biophys. J. 16:143–151
Rousseau, E., Smith, J. S., Henderson, J. S., Meissner, G. 1986. Single channel and45Ca2+ flux measurements of the cardiac sarcoplasmic reticulum calcium channel.Biophys. J. 50:1009–1014
Rousseau, E., Smith, J. S., Meissner, G. 1987. Ryanodine modifies conductance and gating behavior of Ca2+ release channel.Am. J. Physiol. 253:C364-C368
Sachs, F., Neil, J., Bakakati, N. 1982. The automated analysis of data from single ionic channels.Pfluegers Arch. 395:331–340
Schwarze, W., Kolb, H. A. 1984. Voltage-dependent kinetics of an anionic channel of large unit conductance in macrophage and myotube membranes.Pfluegers Arch. 402:281–291
Smith, J. S., Coronado, R., Meissner, G. 1985. Sarcoplasmic reticulum contains adenine nucleotide activated calcium channels.Nature (London) 316:446–449
Smith, J. S., Coronado, R., Meissner, G. 1986. Single channel measurements of the calcium release channel from skeletal muscle sarcoplasmic reticulum. Activation by Ca2+ and ATP and modulation by Mg2+.J. Gen. Physiol. 8:573–588
Smith, J. S., Coronado, R., Meissner, G. 1988a. Techniques for observing calcium channels from skeletal muscle sarcoplasmic reticulum in planar lipid bilayer.Methods Enzymol. 157:480–489
Smith, J. S., Imagawa, T., Ma, J., Campbell, K. P., Coronado, R. 1988b. Purified ryanodine receptor from rabbit skeletal muscle is the calcium release channel of sarcoplasmic reticulum.J. Gen. Physiol. 92:1–26
Suarez-Isla, B. A., Orozco, C., Heller, P. F., Froehlich, J. P.. 1986. Single calcium channel in native sarcoplasmic reticulum membranes from skeletal muscle.Proc. Natl. Acad. Sci. USA 83:7741–7745
Tanifuji, M., Sokabe, M., Kasai, M. 1987. An anion channel of sarcoplasmic reticulum incorporated into planar lipid bilayers: Single channel behavior and conductance properties.J. Membrane Biol. 99:103–111
Tank, D. W., Miller, C., Webb, W. W. 1982. Isolated-patch recording from liposomes containing functionally reconstituted chloride channels fromTorpedo electroplax.Proc. Natl. Acad. Sci. USA 79:7749–7753
Yamamoto, N., Kasai, M. 1981. Kinetic analysis of the inhibition of anion transport in sarcoplasmic reticulum vesicles by a disulfonic stilbene derivative. Measurement of the change in chloride-diffusion potential by using a fluorescent cyanine dye.J. Biochem. 89:1521–1531
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rousseau, E. Single chloride-selective channel from cardiac sarcoplasmic reticulum studied in planar lipid bilayers. J. Membrain Biol. 110, 39–47 (1989). https://doi.org/10.1007/BF01870991
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01870991