Pressure-Induced Depolymerization of Spindle Microtubules: III. Differential Stability in HeLa Cells

Evidence from light microscopy (principally polarization microscopy) has demonstrated that hydrostatic pressure can reversibly inhibit mitosis by rapidly depolymerizing the spindle fiber microtubules. We have confirmed this finding in ultrastructural studies of mitotic HeLa cells incubated at 37°C a...

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Bibliographic Details
Published in:The Journal of cell biology 1976-05, Vol.69 (2), p.443-454
Main Authors: Salmon, E. D., Goode, Dennis, Maugel, T. K., Bonar, Dale B.
Format: Article
Language:English
Subjects:
Anaphase
Cells
Chromosomes
Chromosomes - ultrastructure
Depolymerization
HeLa Cells
Hydrostatic Pressure
Kinetochores
Metaphase
Microtubules
Microtubules - ultrastructure
Mitosis
Mitotic spindle apparatus
Mother cells
Organoids - ultrastructure
Pressure
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Summary:Evidence from light microscopy (principally polarization microscopy) has demonstrated that hydrostatic pressure can reversibly inhibit mitosis by rapidly depolymerizing the spindle fiber microtubules. We have confirmed this finding in ultrastructural studies of mitotic HeLa cells incubated at 37°C and pressurized at 680 atm (10,000 psi). Although there are many spindle microtubules in the cells at atmospheric pressure, electron micrographs of cells pressurized for 10 min (and fixed while under pressure in a Landau-Thibodeau chamber) show few microtubules. Pressure has a differential effect on the various types of spindle microtubules. Astral and interpolar MTs appear to be completely depolymerized in pressurized cells, but occasional groups of kinetochore fiber microtubules are seen. Surprisingly, the length and density of microtubules of the stem bodies and midbody of telophase cells appear unchanged by pressurization. In cells fixed 10 min after pressure was released, microtubules were again abundant, the density often appearing to be higher than in control cells. Reorganization seems incomplete, however, since many of the microtubules are randomly oriented. Unexpectedly, kinetochores appeared diffuse and were difficult to identify in sections of pressurized cells. Even after 10 min of recovery at atmospheric pressure, their structure was less distinct than in unpressurized cells.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.69.2.443