Abstract
Acetylcholine(ACh)-induced inward current was examined in the rat adrenal chromaffin cell in culture. Cells were voltage clamped with one electrode using the giga-seal technique. The ACh dose-response relationship was determined by applying various concentrations of ACh with the “puffer” method. Although there were several technical difficulties, we found tentative figures as 135 to 480 μM for the dissociation constant and 1.2 to 1.4 for the Hill coefficient. External Cs ions had a depressing effect on the ACh reponse. Cs ions may slowly block ACh receptor channels from outside at the resting state. The ACh response in normal saline recorded with the Cs-filled electrode reversed the polarity at −7.1 mV. When the amplitude of ACh-induced current was plotted against holding membrane potential the curve showed strong rectification at the region more negative than the reversal potential. Between +10 and +30 mV there was a region of a negative slope. We discussed possible mechanisms for this characteristic I–V curve.
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References
Adams PR (1975) An analysis of the dose-response curve at voltageclamped frog endplates. Pflügers Arch 360:145–153
Adams DJ, Dwyer TM, Hille B (1980) The permeability of end plate channels to monovalent and divalent metal cations. J Gen Physiol 75:493–510
Anderson CR, Stevens CF (1973) Voltage clamp analysis of acetylcholine produced end-plate current fluctuations at frog neuromuscular junction. J Physiol 235:655–691
Bezanilla F, Armstrong CM (1972) Negative conductance caused by entry of sodium and cesium ions into the potassium channels of squid axon. J Gen Physiol 60:588–608
Brandt BL, Hagiwara S, Kidokoro Y, Miyazaki S (1976) Action potentials in the rat chromaffin cells and effects of acetylcholine. J Physiol 263:417–439
Catterall WA (1975) Sodium transport by the acetylcholine receptor of cultured muscle cells. J Biol Chem 250:1776–1781
Choi DW (1978) Pharmacological evidence for GABA as a neurotransmitter in dissociated spinal cord cell cultures. Ph. D. Thesis. Department of Pharmacology. Harvard University, Cambridge, Massachusetts
Clapham DE, Neher E (1984) Substance P reduces acetylcholine-induced currents in isolated bovine chromaffin cells. J Physiol 347:255–277
Dionne VE, Steinbach JH, Stevens CF (1978) An analysis of the dose-response relationship at voltage-clamped frog neuromuscular junctions. J Physiol 281:421–444
Douglas WW, Kanno T, Sampson SR (1967) Effects of acetylcholine and other medullary secretagogues on the membrane potential of adrenal chromaffin cells: An analysis employing techniques of tissue culture. J Physiol 188:107–120
Dreyer F, Peper K (1975) Density and dose-response curve of acetylcholine receptors in frog neuromuscular junction. Nature 253:641–643
Fenwick EM, Marty A, Neher E (1982a) A patch-clamp study of bovine chromaffin cells and of their sensitivity to acetylcholine. J Physiol 331:577–597
Fenwick EM, Marty A, Neher E (1982b) Sodium and calcium channels in bovine chromaffin cells. J Physiol 331:599–635
Goldman DE (1943) Potential impedance and rectification in membranes. J Gen Physiol 27:37–60
Hagiwara S, Miyazaki S, Rosenthal NP (1976) Potassium current and the effect of cesium on this current during anomalous rectification of the egg cell membrane of a starfish. J Gen Physiol 67:621–638
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patchclamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Hodgkin AL, Katz B (1949) The effect of sodium ions on the electrical activity of the giant axon on the squid. J Physiol 108:38–77
Horn R, Brodwick MS (1980) Acetylcholine-induced current in perfused rat myoballs. J Gen Physiol 75:297–321
Ishikawa K, Harada E, Kanno T (1977) A quantitative analysis of the influence of external calcium and magnesium concentrations on acetylcholine-induced adrenaline release in rat adrenal gland perfused in an antidromic or orthodromic direction. Jpn J Physiol 27:251–266
Kidokoro Y, Ritchie AK (1980) Chromaffin cell action potentials and their possible role in adrenaline secretion from rat adrenal medulla. J Physiol 307:199–216
Kidokoro Y, Miyazaki S, Ozawa S (1982) Acetylcholine-induced membrane depolarization and potential fluctuations in the rat adrenal chromaffin cell. J Physiol 324:203–220
Kidokoro Y, Hirano T, Ohmori H (1984) Acetylcholine dose-response relationship in the rat adrenal chromaffin cell. International Symposium on the Molecular Biology of Catecholamine Storing Tissues, Colmar, France
Kidokoro Y, Ohmori H, Hirano T (1985) Acetylcholine-activated currents in the rat adrenal chromaffin cell in culture. In: Kobayashi H, Bern HA, Urano A (eds) Neurosecretion and the biology of neuropeptides. Tokyo, Japan, pp 96–101
Latorre R, Miller C (1983) Conduction and selectivity in potassium channels. J Membr Biol 71:11–30
Marry A (1981) Ca-dependent K channels with large unitary conductance in chromaffin cell membrane. Nature 291:497–500
Sine SM, Steinbach JH (1984) Agonists block current through acetylcholine receptor channels. Biophy J 46:277–284
Sine SM, Taylor P (1980) The relationship between agonist occupation and the permeability reponse of the cholinergic receptor revealed by bound cobra α-toxin. J Biol Chem 255:10144–10156
Yavin E, Yavin Z (1974) Attachment and culture of dissociated cells from rat embryo cerebral hemispheres on polylysine coated surface. J Cell Biol 62:540–546
Yellen G (1984) Ionic permeation and blockade in Ca2+-activated K+ channels of bovine chromaffin cells. J Gen Physiol 84:157–186
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Hirano, T., Kidokoro, Y. & Ohmori, H. Acetylcholine dose-response relation and the effect of cesium ions in the rat adrenal chromaffin cell under voltage clamp. Pflugers Arch. 408, 401–407 (1987). https://doi.org/10.1007/BF00581136
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DOI: https://doi.org/10.1007/BF00581136