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Updated: Aug 12, 2026

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
Published on: October 1, 2010
Voltage- and time-dependent chloride currents in chick skeletal muscle cells grown in tissue culture
1Department of Physiology, University of Alberta, Edmonton, Canada.
Abstract:
Membrane chloride currents in chick skeletal muscle cells grown in tissue culture were studied by use of the whole cell variation of the patch electrode voltage clamp technique. Small diameter myoballs were obtained by adding colchicine to the growth media. To isolate the currents through the chloride channels, the currents through the sodium, calcium and potassium channels were minimized. With symmetrical chloride concentrations bathing the membrane, inward currents were activated by depolarizations above -45 mV. Above 0 mV, the currents became outward. The reversal potential for the currents shifted with the chloride concentration gradient in a manner consistent with the Nernst relation, indicating that the currents were predominantly carried by chloride ions. The instantaneous current-voltage relation obtained from tail current data was linear. The relationship between conductance and membrane potential was sigmoid. The conductance activated above -45 mV, increased steeply between -45 and -10 mV and saturated above +20 mV. Over the range of potentials where the conductance was just beginning to activate, the conductance increased e-fold for a 7 mV depolarization. The currents activated with an exponential time course and did not decline during step depolarizations. Tail currents declined slowly as the sum of two exponential components. The currents were reversibly suppressed by 100 microM SITS and were irreversibly suppressed by 10 microM DIDS.
Insights
Chick skeletal muscle cells exhibit chloride currents activated by depolarization. These currents, primarily carried by chloride ions, are modulated by specific channel blockers like SITS and DIDS.
Area of Science:
- Cellular electrophysiology
- Ion channel function
Background:
- Skeletal muscle cells possess various ion channels that regulate membrane potential.
- Understanding chloride channel activity is crucial for muscle cell function.
Purpose of the Study:
- To characterize membrane chloride currents in cultured chick skeletal muscle cells.
- To investigate the properties and regulation of these chloride channels.
Main Methods:
- Whole-cell patch-clamp technique on cultured chick skeletal muscle cells.
- Isolation of chloride currents by minimizing other ion channel conductances.
- Voltage-clamp protocols to study current-voltage relationships and activation kinetics.
Main Results:
- Chloride currents activated at depolarized potentials (> -45 mV) and followed Nernst relation for chloride ions.
- Conductance exhibited a sigmoid relationship with membrane potential, activating above -45 mV.
- Currents showed exponential activation and slow biexponential decay, and were inhibited by SITS and DIDS.
Conclusions:
- Cultured chick skeletal muscle cells possess voltage-gated chloride channels.
- These channels are likely involved in regulating membrane potential and excitability.
- SITS and DIDS act as potent blockers of these chloride currents.
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