Abstract
Using freshly isolated single smooth muscle cells prepared by collegenase treatment, membrane currents were recorded by whole-cell voltage clamp. Intracellular constituents were modified by using an intracellular perfusion technique, i.e., pipette solutions were continuously exchanged from control to test solutions during current recording. In smooth muscle cells, intracellular application of ATP, but not cyclic AMP, enchanced the amplitude of Ca2+ currents and prevented current run-down. In addition, with this stabilization of Ca2+ current recording by ATP, introduction of various chemicals into the cell using the intracellular perfusion technique is useful for investigations of regulation of ion channels in smooth muscle cells.
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References
Bolton TB, Tomita T, Vassort G: Voltage clamp and the measurement of ionic conductances in smooth muscle. In: E Bulbring, AF Brading, AW Jones, T Tomita (eds) Smooth Muscle. Edward Arnold, London, 1981, pp 47–64
Gabella G: Structure of smooth muscle. In: E Bulbling, AF Brading, AW Jones, T Tomita (eds) Smooth Muscle. Edward Arnold, London, 1981, pp 1–46
Walsh JV, Singer JJ: Voltage clamp of single freshly dissociated smooth muscle cells; current-voltage relationship for three currents. Pflugers Archiv, Eur J Physiol 390:207–210, 1981
Kleckner U, Isenberg G: Action potentials and net membrane currents of isolated smooth muscle cells (urinary bladder of the guinea-pig). Pflugers Archiv, Eur J Physiol 405:329–339, 1985
Ohya Y, Terada K, Kitamura K, Kuriyama H: Membrane currents recorded from a fragment of rabbit intestinal smooth muscle cell. Am J Physiol 251:C335-C346, 1986
Bean BP, Sturek M, Puga A, Hermsmeyer K: Calcium channels in muscle cells isolated from rat mesenteric arteries: modulation by dihydropyridine drugs. Circulation Research 59:229–235, 1986
Benham CD, Bolton TB, Lang RJ: Acetylcholine activates in inward current in single mammalian smooth muscle cells. Nature 316:345–347, 1985
Ohya Y, Kitamura K, Kuriyama H: Cellular calcium regulates outward currents in rabbit intestinal smooth muscle cell. Am J Physiol 252:C401–0410, 1987
Hagiwara S: Membrane potential-dependent ion channels. In: Cell Membrane, Phylogenetic and Development Approaches: Distinguished Lecture Series of Society of General Physiologists, vol 3. Raven Press, New York, 1982, pp 5–33
Brown AM, Wilson DL, Tsuda Y: Voltage clamp and internal perfusion with suction-pipette method. In: TG Smith Jr, H Lecar, SJ Redman, PW Gage (eds) Voltage and patch clamping with microelectrodes. American Physiological Society, Maryland, 1985, pp 151–170
Kostyuk PG: Intracellular perfusion of nerve cells and its effects on membrane currents. Physiol Review 64:435–454, 1984
Irisawa H, Kokubun S: Modulation by intracellular ATP and cyclic AMP of the slow inward current in isolated single ventricular cells of the guinea-pig. J Physiol 338:321–337, 1983
Soejima M, Noma A: Mode of regulation of the ACh-sensitive K-channel by muscarinic receptor in rabbit atrial cells Pflugers Archiv, Eur J Physiol 400:424–431, 1984
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ: Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Archiv, Eur J Physiol 391:85–100, 1981
Ohya Y, Kitamura K, Kuriyama H: Modulation of ionic current in smooth muscle balls of the rabbit intestine by intracellularly perfused ATP and cAMP. Pflugers Archiv, Eur J Physiol 408:465–473, 1987
Kameyama M, Hofmann F, Trautwein W: On the mechanism of β-adrenergic regulation of the Ca channel in the guinea-pig heart. Pflugers Archiv, Eur J Physiol 405:285–293, 1985
Hille E: The pH-dependent rate of action of local anesthetics on the node of Ranvier. J Gen Physiol 69:475–496, 1977
Kameyama M, Hescheler J, Hoffmann F, Trautwein W: Modulation of Ca current during the phosphorylation cycle in the guinea pig heart. Pflugers Archiv, Eur J Physiol 407:123–128, 1986
Imanaga I, Kameyama M, Irisawa H: Cell-to-cell diffusion of fluorescent dyes in paired ventricular cells. Am J Physiol 252:H223-H232, 1987
Reuter H: Calcium channel modulation by neurotransmitters, enzymes and drugs. Nature 301:569–574, 1983
Fedulova SA, Kostyuk PG, Veselovsky NS: Calcium channels in the somatic membrane of the rat dorsal root ganglion neurons: effects of cAMP. Brain Research 214:210–214, 1981
Sperelakis N, Wahler GM, Bkaily G: Properties of myocardial calcium slow channels and mechanisms of action of calcium antagonistic drugs. In: AE Shamoo (ed) Current Topics in Membranes and Transport. Vol 25. Academic Press, New York, 1985, pp 43–76
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Ohya, Y., Sperelakis, N. Whole-cell voltage clamp and intracellular perfusion technique on single smooth muscle cells. Mol Cell Biochem 80, 79–86 (1989). https://doi.org/10.1007/BF00231006
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DOI: https://doi.org/10.1007/BF00231006