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      Regulation of caveolin and caveolae by cholesterol in MDCK cells.

      Journal of Lipid Research

      Animals, Blotting, Western, Caveolin 1, Caveolins, Cell Line, Cell Membrane, drug effects, ultrastructure, Cholesterol, metabolism, pharmacology, Dogs, Fluorescent Antibody Technique, Indirect, Gene Expression Regulation, Kidney, Membrane Proteins, genetics, Microscopy, Electron, RNA, Messenger

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          Abstract

          We have examined the expression of caveolin in MDCK cells under conditions that vary cellular cholesterol concentration. Caveolin mRNA levels dropped to one-sixth of control levels after treatment with simvastatin, an inhibitor of cholesterol synthesis, or beta-trimethyl cyclodextrin (CD), a cholesterol sequestering drug. Both simvastatin and CD treatment decreased total cellular cholesterol levels to about 50% of control values. The potent activator of the sterol regulatory element, 25-hydroxycholesterol, showed no direct regulation of caveolin mRNA levels. Caveolin protein concentration was also decreased to 50% of control values in cholesterol-depleted cells, giving rise to a severe attenuation of caveolin expression detected by indirect immunofluorescence labeling. Quantitative electron microscopy showed a total loss of morphologically recognizable invaginated caveolae after these cholesterol depletion treatments. When the number of invaginated caveolae per cell was expressed as a function of the cellular cholesterol content, a threshold phenomenon was observed, suggesting that caveolae only form when the steady state cellular cholesterol is above 50% of control values. These findings indicate that caveolins, and caveolae, may play an important part in cellular cholesterol homeostasis.

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          9507997

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