Volume regulation and the efflux of amino acids from cells in incubated slices of rat cerebral cortex. I. Characteristics of transport mechanisms.

The characteristics of amino acid efflux from pre-loaded cells in incubated slices of rat cerebral cortex have been investigated under basal conditions (isosmotic media, 315 mosmol/kg) and following mild hyposmotic shock (265 mOsmol/kg). Rates of efflux have been correlated with the extent of cell swelling in hyposmotic media. Hyposmolality accelerated the slow phase of cellular efflux of L-aspartate (+ 29%), gamma-aminoisobutyric acid (GABA) (+ 38%), L-glutamate (+ 28%) and glycine (+ 26%). The anion transport inhibitor 4,4'-diisothiocyanatostilbene-2,2'-sulfonic acid (DIDS, 25 or 100 microM) as well as trifluoperazine (TFP, 25 microM), an inhibitor of calmodulin activation, both retarded efflux in hyposmotic media, with associated cell swelling (increase in slice non-inulin space). The effects of DIDS and TFP were not additive. N-Ethylmaleimide (NEM, 100 microM) significantly retarded the efflux of neutral amino acids, with cell swelling: these effects were less pronounced in cells loaded with acidic amino acids. It is concluded that the hyposmotically-activated efflux of carboxylic amino acids, and associated cell swelling limitation, requires calmodulin activation and the presence of free sulfydryl groups.