alpha(2)-adrenergic agonists reduce glutamate release and glutamate receptor-mediated calcium changes in hippocampal slices during hypoxia

The mechanisms by which alpha(2)-adrenergic agonists reduce ischemic brain damage are not clear. In ischemia-vulnerable hippocampal neurons we tested whether alpha(2)-agonists reduce glutamate efflux and glutamate receptor-mediated increase of cytosolic free calcium. Brain slices (300 mu m thick) from rat hippocampus were loaded with fura-2 for measurements of cytosolic free calcium with a microscope fluorometer. Change of cytosolic calcium in CA1 neurons during application of N-methyl-D-aspartate (NMDA) was measured, as were calcium changes during simulated ischemia (hypoxia, NaCN, iodoacetate) or hypoxia plus high glutamate concentration (pO(2) = 25 mmHg, 3 mM glutamate). In other slices, glutamate efflux evoked by anoxia (pO(2) = 25 mmHg, 100 mu M NaCN) was measured. The selective alpha(2)-agonist mivazerol (1 mu M) decreased NMDA receptor-mediated calcium changes in hippocampal CA1 neurons by 28% (p = 0.0079). With hypoxia and 3 mM glutamate, 1 mu M mivazerol reduced early peak calcium changes in CA1 neurons by 57% (p = 0.0007). An alpha(2)-antagonist (rauwolscine, 1 mu M) blocked this. Mivazerol did not reduce the rate of calcium change during simulated ischemia. Clonidine (0.1 mu M), a partial alpha(2)-agonist, decreased glutamate/hypoxia-mediated calcium changes in CA1 (p = 0.01), but 1 mu M clonidine, which stimulates alpha(1)-receptors, did not. Mivazerol decreased hypoxia and KCl-evoked glutamate release by 50% and 75% (p < 0.01), respectively. In addition, 1 mu M mivazerol reduced lactate dehydrogenase leakage rate from brain slices during anoxia by 61% (p = 0.018). Thus, alpha(2)-receptors influence glutamate release, calcium changes, and cell damage in ischemia-vulnerable hippocampal neurons. These effects may contribute to the cerebroprotective actions of alpha(2)-agonists. Copyright (C) 1996 Elsevier Science Ltd.