Stimulation of Xenopus laevis oocyte maturation by methyl-beta-cyclodextrin.
The cholesterol-depleting drug methyl-beta-cyclodextrin (Me-beta-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-beta-CD in a dose-dependent manner. Treatment of oocytes with 50 mM Me-beta-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 [(3)H]cholesterol, 30-90 min of treatment with 5-50 mM Me-beta-CD removed 50%-70% of cell- associated label, and cholesterol depletion was not observed with alpha-cyclodextrin. After long-term (20-23 h) labeling of oocytes with [(3)H]cholesterol, Me-beta-CD treatment resulted in dose- dependent cholesterol depletion in the 5-50 mM range, and 50 mM Me-beta-CD removed approximately 50% of cell-associated label after 9 h. Treatment of oocytes with 5-50 mM Me-beta-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.
Sadler
Susan E
SE
Department of Biological Sciences, University of Denver, Denver, Colorado 80208, USA. ssadler@du.edu
Jacobs
Nicole D
ND
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D009866:Oogenesis / Q000187:drug effects; D011374:Progesterone / Q000494:pharmacology; D015398:Signal Transduction; D014982:Xenopus laevis / Q000502:physiology*; D047392:beta-Cyclodextrins / Q000008:administration & dosage / Q000494:pharmacology*
D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D011374:Progesterone; D002784:Cholesterol