Cytogenetic assays for mitotic poisons. The grasshopper embryo system for volatile liquids

The present study demonstrates the usefulness of grasshopper (Melanoplus sanguinipes) embryos for detecting mitotic arrest and anaphase abnormalities induced by volatile liquids. Eight compounds were tested with this system. Embryos in vivo (intact chorions) were suspended in sealed chambers and the agents, in various amounts, were placed on the bottom of the chambers. The agents, after evaporation, filled the ambient air in which the embryos respired. After a period of exposure, individual embryos were fixed for squash preparations to determine the mitotic index (MI) and the ratio of anaphases to metaphases (A/M). If an agent completely arrested mitosis at metaphase, the A/M ratio would be zero and the MI would be elevated if the agent does not prevent interphase cells from entering mitosis. Two common organic solvents, benzene and toluene, and two common anesthetics, halothane and chloroform, were found to be potent mitotic arrestants in the present system. Colchicine-like mitoses (c-mitoses) were observed in embryos after treatment of the above four agents; all of these agents except toluene were found to induce an accumulation of c-mitoses. Amyl acetate, 95% ethanol, deionized water, and camphor (in 10% ethanol) failed to show mitosis-arresting effects on the embryos. Abnormal anaphases with lagging chromosomes and multipolar spindles were observed in embryos which showed partial metaphase block after exposure to concentrations of benzene lower than that required for a complete block. Benzene, toluene, halothane, and chloroform have been implicated as teratogens and carcinogens, but are negative as mutagens, clastogens, and SCE inducers. Thus, the carcinogenic activity of these compounds appears to result from their action on cell division mechanisms.