Abstract
Membrane currents through the Ca2+ channel were studied in a hybridoma cell line (MAb-7B) constructed by fusion of S194 myeloma cells and splenic B lymphocytes from the mouse. The whole-cell variation of the patch-electrode voltage-clamp technique was used. When [Ca2+]o = 2.5 mM, [Na+]o = 150 mM and [Na+]i = 155 mM, the current reversed from inward to outward at 20.9 +/- 2.4 mV (mean +/- S.D., n = 62). Both inward and outward currents showed voltage-dependent inactivation with the same membrane potential dependence of steady-state inactivation. The decay time constant of the current decreased from about 27 ms at -44 mV to a saturation value of 16 ms at about -20 mV, and remained at this value even when the current became outward. From the above results both the inward and outward currents were considered to flow through Ca2+ channels. The inward current showed no change when the external Na+ was replaced with Cs+ or tetraethylammonium and increased when [Ca2+]o was increased. Also, the reversal potential became more positive with increasing [Ca2+]o with a slope of 29 mV/decade change of [Ca2+]o. Effects of different divalent cations examined at 10 mM concentration showed the reversal potential to become more positive in the order of Mn2+, Sr2+ approximately equal to Ba2+ and Ca2+ whereas the relative maximum amplitudes of peak inward current were 1.0 for Ca2+, 1.24 for Sr2+, 0.99 for Ba2+ and 0.07 for Mn2+. When [Ca2+]o or [Mg2+]o was reduced by chelators, monovalent cations became capable of carrying inward current through the Ca2+ channel. These monovalent currents share common kinetic properties with the Ca2+ current, as judged from the steady-state inactivation and the decay time constant of the current. The monovalent cation current was blocked by divalent cations in a voltage-dependent manner. The half-blocking concentrations of Ca2+ and Mg2+ at -45 mV were 2.0 X 10(-6) M and 3.0 X 10(-5) M respectively. The same voltage-dependent binding mechanism can explain the outward current carried by monovalent cations at large positive potentials at normal Ca2+ concentrations. The suppression of the monovalent currents by Ca2+ and Mg2+ showed different voltage dependences. The suppression by Ca2+ increased and then decreased as the membrane potential was made negative, whereas the suppression by Mg2+ increased monotonically. This difference can be explained by considering the fact the Ca2+ is permeant and Mg2+ is impermeant through the Ca2+ channel.
MeSH Terms
Action Potentials/drug effects
Animals
B-Lymphocytes/physiology
Calcium/metabolism
Cations, Divalent/pharmacology
Cations, Monovalent
Cell Line
Chelating Agents/pharmacology
Electric Conductivity
Hybridomas/cytology
Ion Channels/drug effects,physiology
Membrane Potentials/drug effects
Mice
Chemicals
Cations, Divalent
Cations, Monovalent
Chelating Agents
Ion Channels
Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fukushima Y
Hagiwara S
References (22)
22 references, click to expand
-
Potassium current in clonal cytotoxic T lymphocytes from the mouse.
J Physiol. 1984 Jun;351:645-56
PMID: 6611410
-
Mechanism of ion permeation through calcium channels.
Nature. 1984 May 31-Jun 6;309(5967):453-6
PMID: 6328315
-
Non-selective conductance in calcium channels of frog muscle: calcium selectivity in a single-file pore.
J Physiol. 1984 Aug;353:585-608
PMID: 6090646
-
Variation of calcium current during the cell growth cycle in mouse hybridoma lines secreting immunoglobulins.
J Physiol. 1984 Oct;355:313-21
PMID: 6491993
-
Surface density of calcium ions and calcium spikes in the barnacle muscle fiber membrane.
J Gen Physiol. 1967 Jan;50(3):583-601
PMID: 11526848
-
The permeability of the sodium channel to metal cations in myelinated nerve.
J Gen Physiol. 1972 Jun;59(6):637-58
PMID: 5025743
-
A study of the ion selectivity and the kinetic properties of the calcium dependent slow inward current in mammalian cardiac muscle.
J Physiol. 1977 Jan;264(1):17-47
PMID: 839451
-
Effects of calcium and calcium-chelating agents on the inward and outward current in the membrane of mollusc neurones.
J Physiol. 1977 Sep;270(3):569-80
PMID: 409839
-
Potassium channels as multi-ion single-file pores.
J Gen Physiol. 1978 Oct;72(4):409-42
PMID: 722275
-
Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells.
J Physiol (Paris). 1979;75(5):463-505
PMID: 533865
-
Rubidium block and rubidium permeability of the inward rectifier of frog skeletal muscle fibres.
J Physiol. 1980 Jul;304:415-35
PMID: 7441543
-
Calcium channel.
Annu Rev Neurosci. 1981;4:69-125
PMID: 6261668
-
Free calcium ions in neurones of Helix aspersa measured with ion-selective micro-electrodes.
J Physiol. 1981 Jun;315:531-48
PMID: 6273543
-
Reversal of current through calcium channels in dialysed single heart cells.
Nature. 1982 Jun 10;297(5866):498-501
PMID: 6283359
-
Intracellular metabolism of adenosine 3',5'-cyclic monophosphate and calcium inward current in perfused neurones of Helix pomatia.
Neuroscience. 1982;7(9):2125-34
PMID: 6292777
-
Studies of calcium channels in rat clonal pituitary cells with patch electrode voltage clamp.
J Physiol. 1982 Oct;331:231-52
PMID: 6296367
-
Blocking kinetics of the anomalous potassium rectifier of tunicate egg studied by single channel recording.
J Physiol. 1982 Oct;331:311-31
PMID: 6296368
-
Sodium and calcium channels in bovine chromaffin cells.
J Physiol. 1982 Oct;331:599-635
PMID: 6296372
-
Voltage-gated Ca2+ channel in mouse myeloma cells.
Proc Natl Acad Sci U S A. 1983 Apr;80(8):2240-2
PMID: 6300901
-
Determination of ionic calcium in frog skeletal muscle fibers.
Biophys J. 1983 Jul;43(1):1-4
PMID: 6603872
-
Calcium current activation kinetics in neurones of the snail Lymnaea stagnalis.
J Physiol. 1984 Mar;348:187-207
PMID: 6325671
-
A non-selective cation conductance in frog muscle membrane blocked by micromolar external calcium ions.
J Physiol. 1984 Aug;353:565-83
PMID: 6090645