## Abstract We examined the calmodulin dependence of anion channel activation during hypoβosmotic swelling in rat cerebral astrocytes. Control cells bathed in isoβosmotic (290 mOsm) phosphateβbuffered saline (PBS) and recorded using a patch electrode containing 140 mM KCl increased membrane conduct
Energy-dependent volume regulation in primary cultured cerebral astrocytes
β Scribed by James E. Olson; Raman Sankar; David Holtzman; Adrian James; Deborah Fleischhacker
- Publisher
- John Wiley and Sons
- Year
- 1986
- Tongue
- English
- Weight
- 943 KB
- Volume
- 128
- Category
- Article
- ISSN
- 0021-9541
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β¦ Synopsis
Cell volume regulation and energy metabolism were studied in primary cultured cerebral astrocytes during exposure to media of altered osmolarity. Cells suspended in medium containing % the normal concentration of NaCI (hypoosmotic) swell immediately to a volume 40-50% larger than cells suspended in isoosmotic medium. The cell volume in hypoosmotic medium then decreases over 30 min to a volume approximately 25% larger than cells in isoosmotic medium. In hyperosmotic medium (containing twice the normal concentration of NaCI), astrocytes shrink by 29%. Little volume change occurs following this initial shrinkage. Cells resuspended in isoosmotic medium after a 30 min incubation in hypoosmotic medium shrink immediately to a volume 10% less than the volume of cells incubated continuously in isoosmotic medium. Thus, the regulatory volume decrease (RVD) in hypoosmotic medium involves a net reduction of intracellular osmoles. The RVD is partially blocked by inhibitors of mitochondrial electron transport but is unaffected by an inhibitor of glycolysis or by an uncoupler of oxidative phosphorylation. Inhibition of RVD by these metabolic agents is correlated with decreased cellular ATP levels. Ouabain, added immediately after hypoosmotic induced swelling, completely inhibits RVD, but does not alter cell volume if added after RVD has taken place. Ouabain also inhibits cell respiration 27% more in hypoosmotic medium than in isoosmotic medium indicating that the (Na,K)-ATPase-coupled ion pump is more active in t h e hypoosmotic medium. These data suggest that the cell volume response of astrocytes in hypoosmotic medium involves the net movement of osmoles by a mechanism dependent on cellular energy and tightly coupled to the (Na,K)-ATPase ion pump. This process may be important in the energy-dependent osmoregulation in the brain, a critical role attributed to the astrocyte in vivo.
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