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The processes that participate in clearing increases in [K+]o produced by active neurons include KCl uptake, Na pump stimulation, and spatial buffering. The latter process requires glial cells to carry: 1) inward K+ currents in regions where K+ is elevated at a glial membrane potential more negative than EK; and 2) outward K+ currents at normal K+ and glial membrane potential more positive than EK (Orkand et al: J Neurophysiol 29:788, 1966). Techniques for isolation and culturing glial cells brought new possibilities for studying ionic channels involved in spatial buffering. However, they raised the question of the extent to which the properties of ionic channels are changed due to the process of culturing when glial cells are exposed to an artificial environment and deprived of direct interaction with neurons. We studied potassium currents in glial cells from the frog optic nerve that were cultured for 1-8 days. At 24-48 h, cells exhibited an inwardly rectifying Cs+ blocked current (IK(IN)) that increased in amplitude and shifted its threshold of activation to EK when [K+]o was increased from 3 to 6 or 10 mM. IK(IN), diminished after 3 days in culture and virtually disappeared after 5 days. At 24-48 h, a potassium delayed rectifier current (IKD) was relatively small but became large at 3 days, and was practically the only current present after 5 days. IKD was activated at -8.5 +/- 0.58 mV(SE, n = 48) and 58 +/- 2.2% (SE, n = 48) blocked by 20 mM tetraethylammonium. The results of this study support the idea that the inward rectifying potassium channels (Kir) are responsible for carrying K+ into glial cells whenever [K+]o increases. However, the delayed rectifier potassium channels (KD) cannot provide the pathway for outward K+ current during spatial buffering, and another mechanism must be involved in this process. Our study provides further evidence that culture conditions can greatly influence functional expression of ionic channels in glial cells. |
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