Abstract
This study aimed at testing if, and under which conditions, long-lasting cytosolic calcium responses can be induced in dissociated embryonic brain cells exposed to alpha-amino-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptor agonists. Rat brainstem cells (gestation days 13-14; mean crown-rump lengths 8-11 mm) were mechanically dissociated and loaded with the fluorescent calcium marker Fluo-3 after in vitro delays ranging from 20 min to 6 days. The cells were exposed to various concentrations of AMPA, domoic acid or kainic acid. The evoked fluorescence changes, indicating variations of cytosolic calcium, were recorded and analysed either with a video-microscope or a laser cytometer. Even at the earliest stages, non-desensitizing (or partly desensitizing) calcium responses to AMPA were found. In addition, sequential exposure to AMPA followed either by domoic acid, or by AMPA in the presence of aniracetam, revealed the existence of cells bearing predominantly desensitizing receptors. The non-desensitizing as well as desensitizing response components were blocked by 6,7-dinitroquinoxaline-2,3-dione (DNQX). When the experiments were conducted at 24 degrees C, the cytosolic calcium levels generally returned close to pre-st...Continue Reading
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