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Volume 16, Number 24, Issue of December 15, 1996 pp. 7803-7811
Copyright ©1996 Society for NeuroscienceRelease of [3H]-D-Aspartate from Primary Astrocyte Cultures in Response to Raised External Potassium
Received July 15, 1996; revised Sept. 18, 1996; accepted Sept. 20, 1996.
Eric M. Rutledge1 and Harold K. Kimelberg1, 2 1 Department of Pharmacology and Neuroscience, and 2 Division of Neurosurgery, Albany Medical College, Albany, New York 12208
There are significant Ca2+-independent increases in extracellular glutamate and aspartate during various CNS insults such as ischemia and anoxia. However, the cellular sources of such presumed nonvesicular excitatory amino acid (EAA) release have not been established. To further explore potential mechanisms and sites for EAA release, we studied the release of preloaded [3H]-D-aspartate from primary cultured astrocytes prepared from the cerebral cortices of rat pups. Two phases of release were seen in response to raised KCl. The first phase was small and transient, and the second phase was slower and increased progressively. The initial phase of [3H]-D-aspartate release was greatly enhanced by ouabain pretreatment and was inhibited when astrocytes were preexposed to the EAA transport inhibitor threo-hydroxy
-aspartic acid (THBA). Neither of these manipulations affected the second release component. The second phase of release was inhibited by an anion channel blocker, L-644,711, which is known to inhibit hypotonic swelling-induced release of EAA. Ouabain also resulted in the first phase of release occurring at lower [K+]o. Omission of Ca2+ had no effect on either phase of [3H]-D-aspartate release. These results support the hypothesis that the first component of release in cultured astrocytes is a reversal of the glutamate transporter, and the second component is a result of high KCl-induced swelling. Because marked increases in [K+]o are well established in CNS pathologies such as ischemia, such release may represent a significant source for the increased extracellular EAAs seen in such conditions.
Key words: glutamate; transporter; astroglia; ischemia; potassium; reversal; swelling
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