Sensitivity of cytokinesis to hydrophobic interactions. Chemical induction of bi-and multinucleated cells.
We have tested whether cytokinesis is as sensitive to hydrophobic interactions as karyokinesis, and evaluated the usefulness of the frequency of binucleated cells as end-point. Treating cultured cells for 2 or 24 h, with different lipophilic alcohols and chlorinated hydrocarbons made this possible. Colcemid and cytochalasin B were applied as positive controls for inhibition of karyokinesis and cytokinesis, respectively. Several-fold increases of binucleated cells could be seen with cytochalasin B after 2 h of treatment, while there was no increase with colcemid, which instead blocked cells in prometaphase/metaphase. The solvent acted primarily through hydrophobic interactions. For each solvent, the blocking of cells in prometaphase/metaphase and a minor increase in binucleated cells, were seen at approximately the same concentration; the binucleated cells probably emanated from cells in anaphase/telophase at the start of treatment. We conclude that the spindle function and cleavage show similar sensitivity to hydrophobic interactions. After prolonged treatment, allowing escape from the metaphase block, the solvents induced binucleated and multinucleated cells. By forming the quotient between multinucleated (MULTI) and binucleated (BIN) cells one could distinguish between effects primarily on the spindle or cytokinesis, respectively. All solvents, and a combination of colcemid and cytochalasin B, showed quotients intermediate between those observed with colcemid (high MULTI/BIN) and cytochalasin B (low MULTI/BIN), respectively. Both protocols revealed the same relationship between lowest active concentration and lipophilicity for the solvents, implying that concentration, not dose were of prime importance. The specific inhibitors acted at low concentrations in relation to lipophilicity, clearly demonstrating their chemical mechanisms. This approach can be used for rapid screening of potential aneugens, distinguishing between routes, and when lipophilicity is known, also reveal the principal mechanism of action, i.e. physico-chemical or chemical.
Schultz
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Genetic and Cellular Toxicology, Wallenberg Laboratory, Stockholm University, S-106 91, Stockholm, Sweden.
Onfelt
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A
D000438:Alcohols / Q000494:pharmacology*; D000705:Anaphase; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002251:Carbon Tetrachloride / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects* / Q000502:physiology; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D002725:Chloroform / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D003571:Cytochalasin B / Q000494:pharmacology; D003703:Demecolcine / Q000494:pharmacology; D006843:Hydrocarbons, Chlorinated / Q000494:pharmacology*; D008677:Metaphase; D008938:Mitosis
D000438:Alcohols; D006843:Hydrocarbons, Chlorinated; D003571:Cytochalasin B; D002725:Chloroform; D002251:Carbon Tetrachloride; D003703:Demecolcine