Mitosis arrested by deuterium oxide. Light microscopic, immunofluorescence and ultrastructural characterization.
Dynamics and correctness of mitotic division of PtK1 cells grown in vitro in the presence of deuterium oxide were assessed by morphological criteria. Incubation of PtK1 monolayers for 2 h in media supplemented with 25% to 75% D2O retarded in a dose-dependent manner the interphase/prophase transition, thus substantially decreasing the fraction of prophase cells. The progression of mitosis was also arrested at the metaphase/anaphase transition resulting in an accumulation of prometaphase and metaphase cells. After a prolonged incubation (12-24 h) in 75% D2O, prophase cells reappeared, often displaying a peculiar, "overmatured" pattern of chromosome condensation. In the presence of D2O an "overmatured" prophase cell is able to transform into a prometaphase cell and then directly into a multinucleate/micronucleate cell. Arrested prometaphase and metaphase cells developed restitution nuclei when incubated up to 24 h in the media containing 25% to 75% of heavy water. Immunofluorescence and electron microscopic studies revealed that the treatment with deuterium oxide profoundly disturbed development of the mitotic spindle. On the basis of these observations, a possible mechanism of the antimitotic action of deuterium oxide is discussed. We postulate that centrosomally controlled transformation of the microtubular cytoskeleton into the mitotic spindle is impaired by D2O. This delays the progression of prophase and disturbs congression of chromosomes into a regular metaphase plate.
Lamprecht
J
J
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
Schroeter
D
D
Paweletz
N
N
D000818:Animals; D000970:Antineoplastic Agents; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D003903:Deuterium / Q000494:pharmacology*; D017666:Deuterium Oxide; D005455:Fluorescent Antibody Technique; D008853:Microscopy; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects*; D014404:Tubulin / Q000032:analysis; D014867:Water / Q000494:pharmacology*
D000970:Antineoplastic Agents; D014404:Tubulin; D014867:Water; D003903:Deuterium; D017666:Deuterium Oxide