Real-time visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts.
Using Xenopus egg extracts arrested in interphase or mitosis, we directly observed differences in microtubule dynamics at different stages of the cell cycle. Interphase extracts were prepared from eggs in the first interphase after meiosis. Mitotic extracts were prepared by addition of purified cyclin to interphase extracts. Microtubules were nucleated by the addition of centrosomes and visualized by fluorescence video-microscopy in extracts to which rhodamine-labeled tubulin had been added. We found a striking difference in microtubule dynamics in mitotic versus interphase extracts. Quantitative analysis revealed that the rates of polymerization and depolymerization are similar in interphase and mitosis and that within the spatial and temporal resolution of our experiments the difference in dynamics is due almost entirely to an increase in the frequency of transition from growing to shrinking (catastrophe frequency) in the mitotic extracts.
Belmont
L D
LD
Department of Biochemistry and Biophysics, University of California, San Francisco 94143.
Hyman
A A
AA
Sawin
K E
KE
Mitchison
T J
TJ
D000818:Animals; D002453:Cell Cycle; D002467:Cell Nucleus / Q000648:ultrastructure; D003513:Cycloheximide / Q000494:pharmacology; D003593:Cytoplasm / Q000648:ultrastructure; D005260:Female; D007399:Interphase; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D008938:Mitosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D012235:Rhodamines; D013997:Time Factors; D014404:Tubulin; D014741:Video Recording; D014982:Xenopus laevis
D012235:Rhodamines; D014404:Tubulin; D003513:Cycloheximide