Amino acid current through anion channels in cultured human glial cells.
During volume regulation in hypotonic media, glial cells release a large portion of their amino acids. These amino acid losses appear to be mediated by a diffusion type of transport and a swelling-activated chloride channel seems to be involved. The objective of this project was to provide direct evidence that amino acids could diffuse through a Cl- channel. Using a human glial cell line, Cl- currents activated in hypotonic media were measured in whole-cell patch clamp. To measure the currents produced by amino acids, it was necessary to increase the pH of external solutions to basic values reaching 9.6 and 10.0 to raise the concentration of the anionic form of these amino acids. Introducing external hypotonic media containing high concentrations of amino acids, like glycine, taurine, glutamine and glutamate, it was possible to measure their respective current-voltage curves with NMDG-Cl-filled pipettes. From the reversal potentials, their permeability ratios with respect to chloride were determined. It was found that the low molecular weight amino acids, like glycine, were most permeant, while the larger ones, like glutamine, had a lower permeability with respect to chloride. The amino acids with two carboxyl groups, like glutamate, had a much lower permeability ratio. The reversal potentials for some metabolites, like lactate and malate were also measured for comparison. These results demonstrate that amino acids can diffuse through anion channels and that activation of these channels in pathological conditions could be at least partly responsible for the observed increase in external amino acids.
Roy
G
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Département de Physiologie, Université de Montréal, Québec, Canada.
D000596:Amino Acids / Q000378:metabolism*; D001693:Biological Transport, Active; D002460:Cell Line; D002463:Cell Membrane Permeability; D018118:Chloride Channels / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007038:Hypotonic Solutions; D017136:Ion Transport; D008564:Membrane Potentials; D009457:Neuroglia / Q000378:metabolism*
D000596:Amino Acids; D018118:Chloride Channels; D002712:Chlorides; D007038:Hypotonic Solutions