Abstract
Critical for karyokinesis in animal cells, is the assembly of a spindle-chromosome-aster complex which serves as the force-generation system for chromosome segregation. Spisula solidissima oocytes offer a unique system for the study of spindle assembly since they can be obtained in large quantities, are naturally synchronized in the cell cycle and can be induced to assemble meiotic spindles within 15 min after parthenogenetic activation. Oocytes contain a large nucleus [germinal vesicle (GV)] and are arrested at the G2/M border of meiosis I. No centrosomes or asters have been observed in oocytes at this stage by either electron microscopy or fluorescence microscopy using anti-tubulin antibodies. Observation of live oocytes with polarized light microscopy revealed that within 4-6 min after parthenogenetic activation, asters appeared and invaded the GV as it breaks down (GVBD). 15 min after activation a complete bipolar meiotic spindle was observed.As a first step towards the development of an in vitro biochemical reconstitution system for the study of spindle assembly, concentrated lysates were prepared at various times following parthenogenetic activation and analyzed by phase-contrast microscopy.
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Schedule a callMore From: Proceedings, annual meeting, Electron Microscopy Society of America
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