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Stimulation of Xenopus laevis Oocyte Maturation by Methyl-ß-Cyclodextrin¹

Department of Biological Sciences, University of Denver, Denver, Colorado 80208

The cholesterol-depleting drug methyl-ß-cyclodextrin (Me-ß- CD) was tested for its effects on amphibian oocyte maturation, cholesterol depletion, and low-density membrane recovery. Progesterone-induced oocyte maturation was accelerated by pretreatment of cells with 5–50 mM Me-ß-CD in a dose-dependent manner. Treatment of oocytes with 50 mM Me-ß-CD alone was sufficient to induce germinal vesicle breakdown, stimulate formation of meiotic spindles, and stimulate phosphorylation of mitogen-activated protein kinase over time courses longer than those observed after progesterone treatment. After short-term (30 min) labeling of oocytes with [³H]cholesterol, 30–90 min of treatment with 5–50 mM Me-ß-CD removed 50%–70% of cell- associated label, and cholesterol depletion was not observed with -cyclodextrin. After long-term (20–23 h) labeling of oocytes with [³H]cholesterol, Me-ß-CD treatment resulted in dose- dependent cholesterol depletion in the 5–50 mM range, and 50 mM Me-ß-CD removed approximately 50% of cell-associated label after 9 h. Treatment of oocytes with 5–50 mM Me-ß-CD also decreased recovery of low-density membrane by detergent- free sucrose gradient centrifugation. These results implicate cholesterol and low-density membrane domains in the signaling mechanisms leading to germinal vesicle breakdown in amphibian oocytes.

¹ This work was supported by NIH grant 1 R15 GM60922 to S.E.S. Preliminary data from these studies were presented as a poster at the 2001 annual meeting of the American Society for Cell Biology.

² Correspondence: Susan E. Sadler, University of Denver, Department of Biological Sciences, 2101 East Wesley, University Park, Denver, CO 80208. FAX: 303 871 3471; ssadler{at}du.edu

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