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 microns thick) from rat hippocampal were located 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) of hypoxia plus high glutamate concentration (pO2 = 25 mmHg, 3 mM glutamate). In order slices, glutamate efflux evoked by anoxia (pO2 = 25 mmHg, 100 microM NaCN) was measured. The selective alpha 2-agonist mivazerol (1 microM) decreased NMDA receptor-mediated calcium changes in hippocampal CA1 neurons by 28% (p = 0.0079). With hypoxia and 3 mM glutamate, 1 microM mivazerol reduced early peak calcium changes in CA1 neurons by 57% (p = 0.0007). An alpha 2-antagonist (rauwolscine, 1 microM) blocked this. Mivazerol did not reduce the rate of calcium change during simulated ischemia. Clonidine (0.1 microM), a partial alpha 2-agonist, decrease glutamate/hypoxia-mediated calcium changes in CA1 (p = 0.01), but 1 microM clonidine, which stimulates alpha 1-receptors, did not. Mivazerol decreased hypoxia and KCl1-evoked glutamate release by 50% and 75% (p < 0.01), respectively. In addition, 1 microM 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.
Department of Anesthesia, University of California at San Francisco, 94143, USA. bickler@jemo.ucsf.edu
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Hansen
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003000:Clonidine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D018698:Glutamic Acid / Q000378:metabolism*; D006624:Hippocampus / Q000187:drug effects*; D000860:Hypoxia / Q000378:metabolism*; D007093:Imidazoles / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000316:Adrenergic alpha-Agonists; D007093:Imidazoles; D018698:Glutamic Acid; D003000:Clonidine; D002118:Calcium; C087969:mivazerol