Related Experiment Videos
Spindle microtubule dynamics: modulation by metabolic inhibitors
1Department of Zoology, University of Massachusetts, Amherst 01003.
Cell Motility and the Cytoskeleton
|January 1, 1988
Summary
Cellular microtubule dynamics, essential for cell division, require adenosine triphosphate (ATP) for tubulin subunit turnover. Inhibiting ATP production significantly impairs microtubule recovery and organization.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Spindle microtubules exhibit high dynamism, with tubulin subunits constantly exchanging.
- Fluorescence recovery after photobleaching (FRAP) quantifies tubulin turnover in living cells.
Purpose of the Study:
- To investigate the energy requirements for tubulin turnover in living cells.
- To elucidate the role of ATP in regulating microtubule dynamics.
Main Methods:
- Microinjection of fluorescent tubulin into cells.
- Measurement of tubulin exchange using FRAP.
- ATP depletion using sodium azide and 2-deoxyglucose (Az/DOG).
- Assessment of spindle morphology via microscopy and immunofluorescence.
Main Results:
- ATP depletion significantly reduced the rate and extent of tubulin recovery in FRAP experiments.
- Glycolysis partially supported microtubule turnover, but less efficiently than normal ATP production.
- Extended ATP depletion altered microtubule organization and increased resistance to nocodazole disassembly.
Conclusions:
- Microtubule dynamics and tubulin turnover are ATP-dependent processes.
- There is a link between microtubule organization, dynamics, and cellular energy status.