The role of microfilaments in early meiotic maturation of mouse oocytes.
Mouse oocyte microfilaments (MF) were perturbed by depolymerization (cytochalasin B) or stabilization (jasplakinolide) and correlated meiotic defects examined by confocal microscopy. MF, microtubules, and mitochondria were vitally stained; centrosomes (gamma-tubulin), after fixation. MF depolymerization by cytochalasin in culture medium did not affect central migration of centrosomes, mitochondria, or nuclear breakdown (GVBD); some MF signal was localized around the germinal vesicle (GV). In maturation-blocking medium (containing IBMX), central movement was curtailed and cortical MF aggregations made the plasma membrane wavy. Occasional long MF suggested that not all MF were depolymerized. MF stabilization by jasplakinolide led to MF aggregations throughout the cytoplasm. GVBD occurred (unless IBMX was present) but no spindle formed. Over time, most oocytes constricted creating a dumbbell shape with MF concentrated under one-half of the oocyte cortex and on either side of the constriction. In IBMX medium, the MF-containing half of the dumbbell over time sequestered the GV, MF, mitochondria, and one to two large cortical centrosomes; the non-MF half appeared empty. Cumulus processes contacted the oocyte surface (detected by microtubule content) and mirrored MF distribution. Results demonstrated that MF play an essential role in meiosis, primarily through cortically mediated events, including centrosome localization, spindle (or GV) movement to the periphery, activation of (polar body) constriction, and establishment of oocyte polarity. The presence of a cortical "organizing pole" is hypothesized.
Calarco
Patricia G
PG
Anatomy Department, Box 0452, University of California, San Francisco, 513 Parnassus Ave, San Francisco, CA 94143, USA. calarco@itsa.ucsf.edu
D015056:1-Methyl-3-isobutylxanthine; D008841:Actin Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D000818:Animals; D002478:Cells, Cultured; D008540:Meiosis; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000166:cytology* / Q000648:ultrastructure; D013194:Staining and Labeling
D015056:1-Methyl-3-isobutylxanthine