Summary
Using the method of dehydration and rehydration, rough endoplasmic reticulum (RER) vesicles, isolated by differential centrifugation, can be enlarged to giant liposomes with diameters ranging from 5 to 200 μm. Patch-clamp studies on these giant RER liposomes revealed the existence of a channel with a mean conductance of 260±7 pS (n=23; 140 mmol/liter KCl on both sides). The channel is about four times more permeable for Cl− than for K+. Its activity is strongly voltage regulated. At low potentials (±20 mV) the channel is predominantly in its open state with an open probability near 1.0, whereas it closes permanently at high positive and negative voltages (±70 mV). The channel activity is not influenced by changing the free Ca2+ concentration from 1 mmol/liter to less than 10−9 mol/liter on either side, and is also not affected by typical Cl−-channel blockers like diphenylamine-2-carboxylate (DPC, 1 mmol/liter) or 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid (SITS, 1 mmol/liter). Another chloride channel with a singlechannel conductance of 79±6 pS (n=4) was less frequently observed. In the potential range of −80 to +40 mV this channel displayed no voltage-dependent gating. We assume that these anion channels are involved in the maintenance of electroneutrality during Ca2+ uptake in the RER.
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Bayerdörffer, E., Streb, H., Eckhardt, L., Haase, W., Schulz, I. 1984. Characterization of calcium uptake into rough endoplasmic reticulum of rat pancreas.J. Membrane Biol. 81:69–82
Blatz, A.L., Magleby, K.L. 1983. Single voltage-dependent chloride-selective channels of large conductance in cultured rat muscle.Biophys. J. 43:237–241
Colombini, M. 1983. Purification of VDAC (voltage-dependent anion-selective channel) from rat liver mitochondria.J. Membrane Biol. 74:115–121
Criado, M., Keller, B.U. 1987. A membrane fusion strategy for single-channel recordings of membranes usually non-accessible to patch-clamp pipette electrodes.FEBS Lett. 224:172–176
Fox, J.A. 1987. Ion channel subconductance states.J. Membrane Biol. 97:1–8
Gögelein, H., Greger, R. 1986. A voltage-dependent ionic channel in the basolateral membrane of late proximal tubules of the rabbit kidney.Pfluegers Arch. 407:S142-S148
Gögelein, H., Greger, R. 1987. Properties of single K+ channels in the basolateral membrane of rabbit proximal straight tubules.Pfluegers Arch. 410:288–295
Gögelein, H., Koepsell, H. 1984. Channels in planar bilayers made from commercially available lipids.Pfluegers Arch. 401:433–434
Gray, P.T., Bevan, S., Ritchie, J.M. 1984. High conductance anion-selective channels in rat cultured Schwann cells.Proc. R. Soc. London B 221:395–409
Hamill, O.P., Marty, A., Neher, E., Sakman, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.Pfluegers Arch. 391:85–100
Hanrahan, J.W., Alles, W.P., Lewis, S.A. 1985. Single anionselective channels in basolateral membrane of a mammalian tight epithelium.Proc. Natl. Acad. Sci. USA 82:7791–7795
Imamura, K., Schulz, I. 1985. Phosphorylated intermediate of (Ca2++K+)-stimulated Mg2+-dependent transport ATPase in endoplasmic reticulum from rat pancreatic acinar cells.J. Biol. Chem. 260:11339–11347
Keller, B.U., Hedrich, R., Vaz, W.L.C., Criado, M. 1988a. Single channel recordings of reconstituted ion channel proteins: An improved technique.Pfluegers Arch. 411:94–100
Keller, B.U., Kleineke, J., Criado, M., Söling, H.D. 1988b. Single channel recordings from endoplasmic reticulum membranes of rat liver: A patch clamp study on giant vesicles.Pfluegers Arch. 411:R105
Kemmer, T.P., Bayerdörffer, E., Will, H., Schulz, I. 1987. Anion dependence of Ca2+ transport and (Ca2++K+)-stimulated Mg2+-dependent transport ATPase in rat pancreatic endoplasmic reticulum.J. Biol. Chem. 262:13758–13764
Kolb, H.A., Brown, C.D.A., Murer, H. 1985. Identification of a voltage-dependent anion channel in the apical membrane of a Cl−-secretory epithelium (MDCK).Pfluegers Arch. 403:262–265
Kolb, H.A., Ubl, J. 1987. Activation of anion channels by zymosan particles in membranes of peritoneal macrophages.Biochim. Biophys. Acta 89:239–246
Krouse, M.E., Schneider, G.T., Gage, P.W. 1986. A large anionselective channel has seven conductance levels.Nature (London) 319:58–60
Lindau, M., Fernandez, J.M. 1986. A patch-clamp study of histamine-secreting cells.J. Gen. Physiol. 88:349–368
Maruyama, Y., Petersen, O.H. 1982. Single-channel currents in isolated patches of plasma membrane from basal surface of pancreatic acini.Nature (London) 299:159–161
Montal, M. 1987. Reconstitution of channel proteins from excitable cells in planar lipid bilayer membranes.J. Membrane Biol. 98:101–115
Muallem, S., Schoeffield, M., Pandol, S., Sachs, G. 1985. Inositol trisphosphate modification of ion transport in rough endoplasmic reticulum.Proc. Natl. Acad. Sci. USA 82:4433–4437
Mueller, P., Chien, T.F., Rudy, B. 1983. Formation and properties of cell-size lipid bilayer vesicles.Biophys. J. 44:375–381
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
Ochs, D.L., Korenbrot, J.I., Williams, J.A. 1983. Intracellular free calcium concentrations in isolated pancreatic acini: Effects of secretagogues.Biochem. Biophys. Res. Commun. 117:122–128
Schmid, A., Gögelein, H., Kemmer, T.P., Schulz, I. 1988. Anion channels in large liposomes made from rat pancreatic endoplasmic reticulum vesicles.Pfluegers Arch. 411:R104
Schulz, I. 1980. Messenger role of calcium in function of pancreatic acinar cells.Am. J. Physiol. 239:G335-G347
Schwarze, W., Kolb, H.A. 1984. Voltage-dependent kinetics of an anion channel of large unit conductance in macrophages and myotube membranes.Pfluegers Arch. 402:281–291
Shah, J., Pant, H.C. 1988. Potassium-channel blockers inhibit inositol trisphosphate-induced calcium release in the microsomal fractions isolated from the rat brain.Biochem. J. 250:617–620
Soejima, M., Kokubun, S. 1988. Single anion-selective channel and its ion selectivity in the vascular smooth muscle cell.Pfluegers Arch. 411:304–311
Sorgato, C., Keller, B.U., Stühmer, W. 1987. Patch-clamping of the inner mitochondrial membrane reveals a voltage-dependent ion channel.Nature (London) 330:498–500
Streb, H., Bayerdörffer, E., Haase, W., Irvine, R.F., Schulz, I. 1984. Effect of inositol-1,4,5-trisphosphate on isolated subcellular fractions of rat pancreas.J. Membrane Biol. 81:241–253
Streb, H., Irvine, R.F., Berridge, M.J., Schulz, I. 1983. Release of Ca2+ from a nonmitochondrial intracellular store in pancreatic acinar cells by inositol-1,4,5-trisphosphate.Nature (London) 306:67–69
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
Tedeschi, H., Mannella, C.A., Bowman, C.L. 1987. Patch clamping the outer mitochondrial membrane.J. Membrane Biol. 97:21–29
Wangemann, P., Wittner, M., DiStefano, A., Englert, H.C., Lang, H.J., Schlatter, E., Greger, R. 1986. Cl−-channel blockers in the thick ascending limb of the loop of Henle: Structure activity relationship.Pfluegers Arch. 407:S128-S141
Woll, K.H., Leibowitz, M.D., Neumcke, B., Hille, B. 1987. A high-conductance anion channel in adult amphibian skeletal muscle.Pfluegers Arch. 410:632–640
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Schmid, A., Gögelein, H., Kemmer, T.P. et al. Anion channels in giant liposomes made of endoplasmic reticulum vesicles from rat exocrine pancreas. J. Membrain Biol. 104, 275–282 (1988). https://doi.org/10.1007/BF01872329
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DOI: https://doi.org/10.1007/BF01872329