Mechanisms of regulatory volume decrease in UC-11MG human astrocytoma cells.
Swelling of astrocytes commonly occurs after cerebral ischemia and other brain injuries. Because these cells constitute 20-25% of human brain volume, their swelling is a major factor in the morbidity and mortality associated with cerebral edema. Many cells, including astrocytes, resist or reverse the tendency to swell by activating transport pathways that lead to a regulatory volume decrease. Here we report the results of studies designed to elucidate the mechanisms of the regulatory volume decrease that occurs after astrocytes are swollen by exposure to hypotonic medium. Using UC-11MG cells, a well-characterized, human, astrocytoma-derived line, we observed an increase in membrane permeability to both K+ and Cl- during regulatory volume decrease, consistent with a net loss of these ions. Neither the increase in K+ exit nor the regulatory volume decrease was affected by bumetanide, an inhibitor of anion-cation cotransport. On the other hand, the increased K+ efflux, as well as the regulatory volume decrease, was blocked by Gd3+, suggesting a putative role of stretch-activated cationic channels in the process of volume regulation. Although increases in intracellular free Ca2+ were also observed during hypotonic treatment, they occurred well after the onset of the regulatory volume decrease. Furthermore, the regulatory volume decrease was not affected by blocking the intracellular free Ca2+ increase with dimethyl 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or by removal of extracellular Ca2+. These results indicate that the regulatory volume decrease in UC-11MG cells may involve stretch-activated channels that operate independently of changes in intracellular free Ca2+.
Medrano
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Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati, Ohio 45267-0524.
Gruenstein
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D001253:Astrocytes / Q000502:physiology*; D001254:Astrocytoma / Q000503:physiopathology*; D002034:Bumetanide / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D005682:Gadolinium / Q000494:pharmacology; D006801:Humans; D007038:Hypotonic Solutions; D007424:Intracellular Fluid / Q000502:physiology*; D007700:Kinetics; D009994:Osmolar Concentration; D012413:Rubidium / Q000378:metabolism; D019789:Tetraethylammonium; D013757:Tetraethylammonium Compounds / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014508:Urea / Q000378:metabolism
D002712:Chlorides; D007038:Hypotonic Solutions; D013757:Tetraethylammonium Compounds; D002034:Bumetanide; D019789:Tetraethylammonium; D014508:Urea; D005682:Gadolinium; D012413:Rubidium; D002118:Calcium