Counter-transport of potassium by the glutamate uptake carrier in glial cells isolated from the tiger salamander retina.
1. To investigate the transport of potassium on the glutamate uptake carrier, the glutamate uptake current in isolated retinal Müller cells was monitored by whole-cell clamping, while measuring changes of potassium concentration outside the cells ([K+]o) with an ion-sensitive microelectrode. 2. Activating glutamate uptake led to an accumulation of potassium outside the cells, consistent with the hypothesis, based on less direct evidence, that the glutamate uptake carrier transports potassium out of the cell. 3. The glutamate-evoked rise of [K+]o showed the pharmacology and sodium dependence of glutamate uptake. 4. The rise in [K+]o was proportional to the uptake current flowing between 0 and -80 mV, implying that the ratio of K+ transported to charge transported by the uptake carrier is constant over this voltage range. The K+ to charge transport ratio was the same for uptake of D-aspartate and L-glutamate. 5. By clamping cells with pipettes containing solutions of different [K+], the dependence of the glutamate and aspartate uptake currents on intracellular [K+] was determined. L- and D-aspartate transport showed a smaller maximum uptake current (Imax), and a smaller apparent Michaelis constant (Km) for activation by intracellular K+, than did L-glutamate transport. The ratio of Imax to Km was the same for these three analogues, a result which can be predicted from simple models of the carrier's operation. 6. Fully activating glutamate uptake in Müller cells in the intact retina would produce a K+ load into the extracellular space of about 0.6 mM s-1. Suppression of glutamate release from photoreceptors by light will reduce K+ efflux from Müller cells in the outer retina; this may contribute to the light-evoked fall of [K+]o observed in the outer retina, and thus contribute to shaping the electroretinogram.
Amato
A
A
Department of Physiology, University College London, UK.
Barbour
B
B
Szatkowski
M
M
Attwell
D
D
D027322:Amino Acid Transport System X-AG; D000818:Animals; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000378:metabolism*; D004594:Electrophysiology; D050491:Glutamate Plasma Membrane Transport Proteins; D018698:Glutamic Acid / Q000378:metabolism; D017736:Ion-Selective Electrodes; D007814:Larva; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism*; D012160:Retina / Q000187:drug effects / Q000378:metabolism*; D012964:Sodium / Q000502:physiology; D027981:Symporters; D014562:Caudata
D027322:Amino Acid Transport System X-AG; D002352:Carrier Proteins; D050491:Glutamate Plasma Membrane Transport Proteins; D027981:Symporters; D018698:Glutamic Acid; D012964:Sodium; D011188:Potassium