Glutamate uptake into nerve and glial cells usually functions to keep the extracellular glutamate concentration low in the central nervous system. But one component of glutamate release from neurons is calcium-independent, suggesting a non-vesicular release that may be due to a reversal of glutamate uptake. The activity of the electrogenic glutamate uptake carrier can be monitored by measuring the membrane current it produces, and uptake is activated by intracellular potassium ions. Here we report that raising the potassium concentration around glial cells evokes an outward current component produced by reversed glutamate uptake. This current is activated by intracellular glutamate and sodium, inhibited by extracellular glutamate and sodium, and increased by membrane depolarization. These results demonstrate a non-vesicular mechanism for the release of glutamate from glial cells and neurons. This mechanism may contribute to the neurotoxic rise in extracellular glutamate concentration during brain anoxia.
The cerebellar cortex contains huge numbers of synapses between granule cells and Purkinje cells. These synapses are thought to be a major storage site for information required to execute coordinated movements. To obtain a quantitative description of this connection, we recorded unitary synaptic responses between granule cell and Purkinje cell pairs in adult rat cerebellar slices. Our results are consistent with parallel fiber-->Purkinje cell synapses having high release probabilities and modest paired pulse facilitation. However, a wide range of response amplitudes was observed. Indeed, we detected many fewer parallel fiber connections (7% of the granule cells that were screened) than expected (54%), leading us to suggest that up to 85% of parallel fiber-->Purkinje cell synapses do not generate detectable electrical responses. We also investigated the possible role of granule cell ascending axons by recording granule cells near the Purkinje cell. A high proportion (up to 50%) of local granule cells generated detectable synaptic responses. However, most of these connections were indistinguishable from parallel fiber connections, suggesting that powerful ascending axon connections are rare. The existence of many very weak synapses would provide a mechanism for Purkinje cells to extract information selectively from the mass provided by parallel fibers.
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