Potassium activation of the Na,K-pump in isolated brain microvessels and synaptosomes

Brain capillary endothelial cells play an important role in ion homeostasis of the brain through the transendothelial transport of Na and K. Since little is known about the regulation of ion transport in these cells, we determined the effect of extracellular potassium concentration ([K]o) on the kinetics of the Na,K-pump in isolated cerebral microvessels using both K uptake and Na efflux as measures of pump activity. In addition, we studied K activation of K uptake into synaptosomes under similar conditions to compare this neuronal system to the capillary. When microvessels were preloaded with 22Na by 30 min incubation in K-free buffer, efflux of 22Na into buffer with varying [K]o was dependent on [K]o and inhibited by 7 mM ouabain. This activation of Na efflux was half maximal at 4.2 mM [K]. Ouabain-sensitive K uptake was also half maximally stimulated by a similar [K] in both Na loaded and non-loaded microvessels. In contrast, K uptake into synaptosomes was half maximal at 0.47 mM K. These results demonstrate that both active Na efflux and K uptake into microvessels in vitro are dependent on [K]o in the physiological range. In contrast, synaptosomal K uptake is near maximal at 3 mM K. This suggests that increases in brain [K]o may stimulate ion transport across the cerebral capillary, but will have little effect on Na,K-pump activity in neurons.