Membrane currents and cytoplasmic sodium transients generated by glutamate transport in Bergmann glial cells.
Effects of glutamate and kainate (KA) on Bergmann glial cells were investigated in mouse cerebellar slices using the whole-cell configuration of the patch-clamp technique combined with SBFI-based Na(+) microfluorimetry. L-glutamate (1 mM) and KA (100 microM) induced inward currents in Bergmann glial cells voltage-clamped at -70 mV. These currents were accompanied by an increase in intracellular Na+ concentration ([Na+](i)) from the average resting level of 5.2 +/- 0.5 mM to 26 +/- 5 mM and 33 +/- 7 mM, respectively. KA-evoked signals (1) were completely blocked in the presence of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), an antagonist of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/KA ionotropic glutamate receptors; (2) reversed at 0 mV, and (3) disappeared in Na+ -free, N-methyl-D-glucamine (NMDG+)-containing solution, but remained almost unchanged in Na+ -free, Li+ -containing solution. Conversely, L-glutamate-induced signals (1) were marginally CNQX sensitive (approximately 10% inhibition), (2) did not reverse at a holding potential of +20 mV, (3) were markedly suppressed by Na+ substitution with both NMDG+ and Li+, and (4) were inhibited by D,L-threo-beta-benzyloxyaspartate. Further, D-glutamate, L -, and D-aspartate were also able to induce Na+ -dependent inward current. Stimulation of parallel fibres triggered inward currents and [Na+](i) transients that were insensitive to CNQX and MK-801; hence, we suggested that synaptically released glutamate activates glutamate/Na+ transporter in Bergmann glial cells, which produces a substantial increase in intracellular Na+ concentration.
Kirischuk
Sergei
S
Max-Delbruck-Centre for Molecular Medicine, Robert-Rossle-Strasse 10, 13122 Berlin-Buch, Germany.
Kettenmann
Helmut
H
Verkhratsky
Alexei
A
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D027322:Amino Acid Transport System X-AG / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D001224:Aspartic Acid / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004558:Electric Stimulation; D004594:Electrophysiology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology; D007608:Kainic Acid / Q000494:pharmacology; D008094:Lithium / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008815:Mice, Inbred Strains; D008954:Models, Biological; D009412:Nerve Fibers / Q000502:physiology; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D012964:Sodium / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger / Q000378:metabolism
D027322:Amino Acid Transport System X-AG; D018691:Excitatory Amino Acid Antagonists; D019831:Sodium-Calcium Exchanger; C120673:benzyloxyaspartate; D001224:Aspartic Acid; C047635:N-methylglucosamine; D018698:Glutamic Acid; D016291:Dizocilpine Maleate; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D008094:Lithium; D012964:Sodium; D005944:Glucosamine; D007608:Kainic Acid; D002118:Calcium