Abstract
The effects of protein kinase C (PKC) activation by phorbol ester on intracellular Ca 2+ concentration ([Ca 2+] i) and membrane currents in human microglia grown in culture were investigated. Treatment of microglia with phorbol myristate acetate (PMA) resulted in a large increase in [Ca 2+] i in cells loaded with fura-2. The increased levels of [Ca 2+] i were not altered following removal of the phorbol ester. In Ca 2+-free medium, application of PMA did not increase [Ca 2+] i. In addition, PMA application in standard Ca 2+ -solution containing lanthanum (1.8 mM) had no effect on the microglial response to PMA, suggesting that the phorbol ester actions were due to transmembrane influx of Ca 2+ but not through voltage-gated Ca 2+ channels. Whole-cell patch clamp measurements demonstrated that PMA potentiated an outward K + current and inhibited an inward rectifier K + current. This study is the first demonstration that PKC activation by phorbol ester leads to increased intracellular [Ca 2+] and changes in membrane currents in human microglia.
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