Abstract
(1) Whole-cell and single channel recording techniques have been applied to smooth muscle cells isolated from guinea-pig taenia coli to examine whether multiple types of Ca channels exist. (2) Whole-cell recordings under physiological Ca concentration (1.8 mM) revealed two current components with fast and slow inactivating kinetics. The fast inactivating component was present when cells were held at very negative potentials (−80 mV). It was insensitive to the dihydropyridine (DHP) derivative, nifedipine. In contrast, the slow inactivating component was present at less negative holding potentials. It was blocked by nifedipine. (3) The two current components were found to have closely similar voltage dependencies for activation. (4) These results suggest that the fast inactivating decay of the Ca current was mediated not only by the entry of Ca into the cell but also by a voltage-dependent process via a different type of Ca channel with fast inactivating kinetics. (5) Recordings from cell-attached membrane patches with 100 mM external Ba clearly showed the existence of multiple types of Ca channels with different conductances. (6) The large conductance channels (30 pS) activated at more positive potentials (0 mV) and their averaged current decayed much more slowly. The DHP Ca antagonist, nifedipine, inhibited the large conductance channels increasing the proportion of blank sweeps and reducing the averaged current. On the other had, the DHP Ca-agonist, BayK 8644, increased the averaged current by increasing the mean open-times of the large conductance channels. The presence of micromolar Cd in the patch pipettes produced a flickering block of the large conductance channels. (7) The small conductance channels (7 pS) activated at more negative potentials (−40 mV ∼30 mV) and the averaged current decayed rapidly within 100 ms. They showed no sensitivity to nifedipine and Cd ions. (8) In summary, we have identified at least two distinct types of Ca channels with different conductances, different pharmacological sensitivities, and different voltage- and time-dependent kinetics in smooth muscle cells isolated from guinea-pig taenia coli. The large and small conductance channels could be classified asl- andt-type Ca channels, respectively, which have been described in neuronal and heart cells. The contribution of those channels to the macroscopic component of whole-cell Ca current is also discussed.
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References
Aaronson PI, Benham CD, Bolton TB, Hess P, Lang RJ, Tsien RW (1986) Two types of single-channel and whole-cell calcium or barium currents in single smooth muscle cells of rabbit ear artery and the effects of noradrenaline. J Physiol 377:36P
Amedee BT, Mironneau C, Mironneau J (1987) The calcium channel current of pregnant rat myometrial cells in short-term primary culture. J Physiol 392:253–272
Armstrong D, Eckert R (1987) Voltage-activated calcium channels that must be phosphorylated to respond to membrane depolarization. Proc Natl Acad Sci USA 84:2518–2522
Bean BP (1984) Nitrendipine block of cardiac calcium channels: high affinity binding to the inactivated state. Proc Natl Acad Sci USA 81:6388–6392
Bean BP (1985) Two kinds of calcium channels in canine atrial cells. Differences in kinetics, selectivity and pharmacology. J Gen Physiol 86:1–30
Bean BP, Sturek M, Puga A, Hermsmeyer K (1986) Calcium channels in muscle cells isolated from rat mesenteric arteries: Modulation by dihydropyridine drugs. Circ Res 59:229–235
Benham CD, Hess P, Tries RW (1987) Two types of calcium channels in single smooth muscle cells from rabbit ear artery studied with whole-cell and single-channel recordings. Circ Res 61:10–16
Bonvallet R (1987) A low threshold calcium current recorded at physiological Ca concentrations in single frog aterial cells. Pflügers Arch 408:540–542
Bossu JL, Feltz A, Thomann JM (1985) Depolarization elicits two distinct calcium currents in vertebrate sensory neurons. Pflügers Arch 403:360–368
Carbone E, Lux HD (1984) A low voltage-activated calcium conductance in embryonic chick sensory neurons. Biophys J 46:413–418
Fedulova SA, Kostyuk PG, Veselovsky NS (1985) Two types of Ca channels in the somatic membrane of new-born rat dorsal root ganglion neurons. J Physiol 358:431–446
Fox AP, Nowycky MC, Tsien RW (1987) Single-channel recordings of three types of calcium channels in chick sensory neurons. J Physiol 394:173–200
Friedman ME, Kurtz GS, Kaczorowski GJ, Kats GM, Ruben JP (1986) Two calcium currents in a smooth muscle cell line. Am J Physiol 250:H699-H703
Ganitkevich VYA, Shuba MF, Smirnov SV (1987) Calcium-dependent inactivation of potential-dependent calcium inward current in an isolated guinea-pig smooth muscle cells. J Physiol 392:431–449
Hagiwara S, Ohmori H (1983) Studies of single calcium channel currents in rat clonal pituitary cells. J Physiol 336:649–661
Hagiwara N, Irisawa H, Kameyama M (1988) Contribution of two types of calcium currents to the pacemaker potentials of rabbit sino-atrial node cells. J Physiol 395:233–253
Hamill OP, Marty A, Neher E, Sakamann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Hess P, Lansmann JB, Tsien RW (1984) Different modes of Ca channel gating behaviour favoured by dihydropyridine Ca agonists and antagonists. Nature 311:538–544
Isenberg G, Klöckner U (1982) Calcium tolerant ventricular myocytes prepared by preincubation in a “KB Medium”. Pflügers Arch 395:6–18
Loirand G, Pacaud P, Mironneau C, Mironneau J (1986) Evidence for two distinct calcium channels in rat vascular smooth muscle cells in short-term primary culture. Pflügers Arch 407:566–568
Ma J, Coronado R (1987) Heterogeneity of conductance states in the calcium channel of skeletal muscle transverse tubules. Biophysical J 51:423a
Mitra R, Morad M (1986) Two types of calcium channels in guineapig ventricular myocytes. Proc Natl Acad Sci 83:5340–5344
Nakazawa K, Saito H, Matsuki N (1988) Fast and slowly inactivating activating components of Ca-channel currents and their sensitivities to nicardipine in isolated smooth muscle cells from rat vas deference. Pflügers Arch 411:289–295
Narahashi T, Tsunoo A, Yoshii M (1987) Characterization of two types of calcium channels in mouse neuroblastoma cells. J Physiol 383:231–249
Nilius B, Hess P, Lansman JB, Tsien RW (1985) A novel type of cardiac calcium channel in ventricular cells. Nature 316:443–446
Nowycky MC, Fox AP, Tsien RW (1984) Multiple types of calcium channel in dorsal root ganglion cells distinguished by sensitivity to cadmium and single channel properties. Soc Neurosci 10 (1):526
Nowycky MC, Fox AP, Tsien RW (1985) Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature 316:440–443
Obara K (1984) Isolation and contractile properties of single smooth muscle cells from guinea-pig taenia coli. Jpn J Physiol 34:41–54
Ohmori H, Yoshii M (1977) Surface potential reflected in both gating and permeation mechanisms of sodium and calcium channels of the tunicate egg cell membrane. J Physiol 267:429–463
Ohya Y, Terada K, Kitamura K, Kuriyama H (1986) Membrane currents recorded from a fragment of rabbit intestinal smooth muscle cells. Am J Physiol 251:C335-C346
Ohya Y, Kitamura K, Kuriyama H (1988) Regulation of calcium current by intracellular calcium in smooth muscle cells of rabbit portal vein. Circ Res 62:375–383
Robert L, Hess P, Tsien RW (1988) Cardiac calcium channels in planar lipid bilayers:l-type channels and calcium-permeable channels open at negative membrane potentials. J Gen Physiol 92:27–54
Sanguinetti MC, Kass RS (1984) Regulation of cardiac calcium channel current and contractile activity by the dihydropyridine Bay K 8644 is voltage-dependent. J Mol Cell Cardiol 16:667–670
Sturek M, Hermsmeyer K (1986) Calcium and sodium channels in spontaneously contracting vascular muscle cells. Science 233:475–478
Talventheimo JA, Worley JF, Nelson MT (1986) Purified calcium channels from transverse-tubule membranes incorporated into planar lipid bilayers. J Gen Physiol 88:58a
Wilson DL, Morimoto K, Tsuda Y, Brown AM (1983) Interaction between calcium ions and surface charge as it relates to calcium currents. J Membr Biol 72:117–130
Worley III JF, Deitmer JW, Nelson MT (1986) Single nisoldipinesensitive calcium channels in smooth cells isolated from rabbit mesenteric artery. Proc Natl Acad Sci USA 83:5746–5750
Yoshino M, Nishio A, Yabu H (1988a) Calcium channel modulation by the dihydropyridine derivative Bay K 8644 in mammalian visceral smooth muscle cells. Comp Biochem Physiol 89C:39–43
Yoshino M, Someya T, Nishio A, Yabu H (1988b) Whole-cell and unitary CA channel currents in mammalian intestinal smooth muscle cells: evidence for the existence of two types of Ca channels. Pflügers Arch 411:229–231
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Yoshino, M., Someya, T., Nishio, A. et al. Multiple types of voltage-dependent Ca channels in mammalian intestinal smooth muscle cells. Pflugers Arch. 414, 401–409 (1989). https://doi.org/10.1007/BF00585049
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DOI: https://doi.org/10.1007/BF00585049