Amino acid release during volume regulation by cardiac cells: cellular mechanisms.

Mechanisms of amino acid efflux during volume regulation in hypoosmotically treated isolated rat hearts were studied by collecting the coronary artery perfusate and analysis by high pressure liquid chromatography. Hypoosmotic stress resulted in marked percentage increases in perfusate taurine, aspartate and glutamate levels, smaller increases in phosphoethanolamine, glycine and alanine and non-significant increases in serine and glutamine. Amino acid levels declined during reperfusion with isosmotic perfusate. The anion channel blocker 4-acetamido-4-isothiocyanostilbene-2:2'-disulfonic acid (SITS, 500 microM) significantly reduced hypoosmotic release of taurine, aspartate, glutamate and glycine. Furosemide reduced hypoosmotically-evoked releases of taurine, glycine, alanine and phosphoethanolamine. The polyunsaturated amino acids, arachidonic and linoleic also reduced amino acid efflux. Phospholipase A2 inhibition with 7,7-dimethyleicosadienoic acid (DEDA, 2 microM) reduced osmotically-evoked releases of taurine, aspartate and glutamate. 4-Bromophenacyl bromide (1 microM) inhibited osmotically-evoked release of glutamate and glycine. Combined applications of SITS + DEDA markedly reduced osmotically evoked release of all eight amino acids. Glutamate and aspartate effluxes were not inhibited by the glutamate transport inhibitor dihydrokainic acid (1 mM). These results indicate that the hypoosmotic stress, by inducing cell swelling, can initiate an amino acid efflux as part of a regulatory volume decrease. An opening of anion-permeant channels and phospholipase activation appear to be involved in the regulatory volume decrease phenomenon.