Abstract
The non-contractile aortic smooth muscle cell line A7r5 was used to study the membrane events involved in the effect of vasoconstrictor peptides. Wholecell voltage-clamp and membrane potential recording techniques were used to demonstrate the contribution of an increased Cl− conductance to the late depolarization induced by endothelin-1 and vasopressin. During cell-attached patch recording with N-methyl-d-glucamine in the pipette, bath application of endothelin or vasopressin induced single-channel inward currents in the following minutes. The current/potential (I/V) curve of the most frequently observed channel type — a small conductance Cl− (SCl) channel — reversed near the cell membrane potential and showed a single-channel conductance of 1.8 pS for inward currents. After patch excision in an extracellular solution containing CaCl2 (2 mM), the frequency of SCl channel openings increased. Patch excision in the absence of peptide stimulation also produced this channel activity. Replacement of CaCl2 by a Ca2+ chelator on the intracellular face of a patch reversibly inhibited the channel activity, indicating that these SCl channels are Ca2+-activated Cl− channels. The single-channel I/V characteristic showed outward rectification above +50 mV. An analysis of the gating kinetics of the SCl channel is given. Another channel type was recorded less frequently after peptide stimulation. It had a lower conductance (1.0–1.3 pS) and slower kinetics and was designated a very small conductance Cl}-channel. It is concluded that activation of two types of Cl− channels (at least one of which is Ca2+ dependent) is involved in the late depolarization produced by vasoconstrictor peptides in vascular smooth muscle cells of the aortic cell line A7r5.
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References
Aickin CC, Brading AF (1982) Measurement of intracellular chloride in guinea-pig vas deferens by ion analysis, 36-chloride efflux and micro-electrodes. J Physiol (Lond) 326: 139–154
Aickin CC, Vermuë NA (1983) Microelectrode measurement of intracellular chloride activity in smooth muscle cells of guinea-pig ureter. Pflügers Arch 397: 25–28
Amédée T, Large W, Wang Q (1990) Characteristics of chloride currents activated by noradrenaline in rabbit ear artery. J Physiol (Lond) 428: 501–516
Bader CR, Bertrand D, Schwartz EA (1982) Voltage-activated and calcium-activated currents studied in solitary rod inner segments from the salamander retina. J Physiol (Lond) 331: 253–284
Barish M (1983) A transient calcium-dependent chloride current in the immature Xenopus occyte. J Physiol (Lond) 342: 309–325
Baron A, Pacaud P, Loirand G, Mironneau C, Mironneau J (1991) Pharmacological block of Ca2+-activated Cl− current in rat vascular smooth muscle cells in short term primary culture. Pflügers Arch 419: 553–558
Bolton T (1979) Mechanism of action of transmitters and other substances on smooth muscle. Physiol Rev 59: 607–718
Byrne NG, Large WA (1987) Action of noradrenaline on single smooth muscle cells freshly dispersed from the rat anococcygeus muscle. J Physiol (Lond) 389: 513–525
Byrne NG, Large WA (1988) Membrane ionic mechanisms activated by noradrenaline in cells isolated from the rabbit portal vein. J Physiol 404: 557
Chesnoy-Marchais D (1982) A Cl− conductance activated by hyperpolarization in Aplysia neurones. Nature 299: 359–361
Droogmans G, Callewaert G, Declerck I, Castells R (1991) ATP-induced Ca2+ release and Cl− current in cultured smooth muscle cells from pig aorta. J Physiol (Lond) 440: 623–634
Evans MG, Marty A (1986) Calcium-dependent chloride currents in isolated cells from rat lacrymal glands. J Physiol (Lond) 378: 437–460
Findlay I, Petersen OH (1985) Acetylcholine stimulates a Ca2+-dependent Cl− conductance in mouse lacrimal acinar cells. Pflügers Arch 403: 328–330
Hirata Y, Yoshimi H, Takata S, Watanabe TX, Kumagai S, Nakajima K, Sakakibara S (1988) Cellular mechanism of action by a novel vasoconstrictor endothelin in cultured rat vascular smooth muscle cells. Biochim Biophys Res Commun 154: 868–875
Hodgkin A, Horowicz P (1959) The influence of potassium and chloride ions on the membrane potential of single muscle fibres. J Physiol (Lond) 148: 127–160
Janssen L, Sims S (1992) Acetylcholine activates non-selective cation and chloride conductances in canine and guinea-pig tracheal myocytes. J Physiol (Lond) 453: 197–218
Kimes BW, Brandt BL (1976) Characterization of two putative smooth muscle cell lines from rat thoracic aorta. Exp Cell Res 98: 349–366
Klöckner U, Isenberg G (1991) Endothelin depolarizes myocytes from porcine coronary and human mesenteric arteries through a Ca-activated chloride current. Pflügers Arch 418: 168–175
Knot H, Ree M de, Gähwiler B, Rüegg U (1991) Modulation of electrical activity and of intracellular calcium oscillations of smooth muscle cells by calcium antagonists, agonist and vasopressin. J Cardiovasc Pharmacol 18: S7-S14
Kremer S, Breuer W, Skoreki K (1989) Vasoconstrictor hormones depolarize renal glomerular mesangial cells by activating chloride channels. J Cell Physiol 138: 97–105
Lunevsky VZ, Zherelova OM, Vostrikov IY, Berestovsky GN (1983) Excitation of Characeae cell membranes as a result of activation of calcium and chloride channels. J Membr Biol 72: 43–58
Marty A, Tan YP, Trautmann A (1984) Three types of calcium-dependent channels in rat lacrymal glands. J Physiol (Lond) 357: 293–325
Matthews G, Neher E, Penner R (1989) Chloride conductance activated by external agonists and internal messengers in rat peritoneal mast cells. J Physiol (Lond) 418: 131–144
Mayer ML (1985) A calcium-activated chloride current generates the after-depolarization of rat sensory neurones in culture. J Physiol (Lond) 364: 217–239
Miledi R (1982) A calcium-dependent transient outward current in Xenopus laevis occytes. Proc R Soc Lond [Biol] 215: 491–497
Owen DG, Segal M, Barker JL (1984) A Ca-dependent Cl− conductance in cultured mouse spinal neurones. Nature 311: 567–569
Pacaud P, Loirand G, Lavie JL, Mironneau C, Mironneau J (1989) Calcium-activated chloride current in rat vascular smooth muscle cells in short term primary culture. Pflügers Arch 413: 629–636
Pacaud P, Loirand G, Mironneau C, Mironneau J (1989) Noradrenalin activates a caclium-activated chloride conductance and increases the voltage-dependent calcium current in cultured single cells of rat portal vein. Br J Pharmacol 97: 139–146
Pacaud P, Loirand G, Grégoire G, Mironneau C, Mironneau J (1992) Calcium-dependence of the calcium-activated chloride current in smooth muscle cells of rat portal vein. Pflügers Arch 421: 125–130
Penner R, Matthews G, Neher E (1988) Regulation of calcium influx by second messengers in rat mast cells. Nature 334: 499–504
Rüegg U, Wallnöfer A, Weir S, Cauvin C (1989) Receptoroperated calcium-permeable channels in vascular smooth muscle. J Cardiovasc Pharmacol 14: S49-S58
Soejima M, Kokubun S (1988) Single anion-selective channel and its ion selectivity in the vascular smooth muscle cell. Pflügers Arch 411: 304–311
Takahashi T, Neher E, Sakmann B (1987) Rat brain serotonin receptors in Xenopus oocytes are coupled by intracellular calcium to endogenous channels. Proc Natl Acad Sci USA 84: 5063–5067
Taleb O, Feltz P, Bossu JL, Feltz A (1988) Small-conductance chloride channels activated by calcium on cultured endocrine cells from mammalian pars intermedia. Pflügers Arch 412: 641–646
Van Helden DF (1988) An α-adrenoceptor-mediated chloride conductance in mesenteric veins of the guinea-pig. J Physiol (Lond) 401: 489–501
Van Renterghem C, Romey G, Lazdunski M (1988) Vasopressin modulates the spontaneous electrical activity in aortic cells (line A7r5) by acting on three different types of ionic channels. Proc Natl Acad Sci USA 85: 9365–9369
Van Renterghem C, Vigne P, Barhanin J, Schmid-Alliana A, Frelin C, Lazdunski M (1988) Molecular mechanisms of action of the vasoconstrictor peptide endothelin. Biochem Biophys Res Commun 157: 977–985
Van Renterghem C, Lazdunski M (1992) A small-conductance charybdotoxin-sensitive, apamin-resistant Ca2+-activated K+ channel in aortic smooth muscle cells (A7r5 line and primary culture). Pflügers Arch 420: 417–423
Vigne P, Breittmayer JP, Lazdunski M, Frelin C (1988) The regulation of the cytoplasmic free Ca concentration in aortic smooth muscle cells (A7r5 line) after stimulation by vasopressin and bombesin. Eur J Biochem 176: 47–52
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Van Renterghem, C., Lazdunski, M. Endothelin and vasopressin activate low conductance chloride channels in aortic smooth muscle cells. Pflugers Arch. 425, 156–163 (1993). https://doi.org/10.1007/BF00374516
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DOI: https://doi.org/10.1007/BF00374516