Cadmium is a toxic environmental contaminant that is carcinogenic in humans and laboratory animals. Although the mechanism underlying cadmium carcinogenesis has not yet been determined experimental evidence suggests that the stress-inducible, metal-binding proteins, metallothioneins, may mediate organ specificity. In the present study, four different rodent cell lines (Chinese hamster ovary cells, rat L6 myoblast cells, rat Clone 9 liver cells, and rat TRL 1215 liver cells) were exposed to 0, 1, 5, 10, 50, or 100 microM CdCl2 and monitored for evidence of direct DNA damage. A microfiltration assay was used to measure DNA strand breaks and a filter-binding assay was used to measure DNA-protein crosslinks, two lesions that have been associated with cadmium exposure and may mediate genotoxicity of the metal. Although variability in sensitivity to DNA damage was evident between the different cell lines, in all of the cell lines tested, increases in DNA damage were observed only at cadmium doses that completely arrested cell growth. In addition, in three of the four cell lines tested, induction of metallothionein had no substantial protective effect against cadmium-induced cytotoxicity or genotoxicty. While protection against cadmium-induced DNA strand breakage with metallothionein preinduction was observed in the TRL 1215 rat liver cells, metallothionein preinduction did not protect against cadmium-induced DNA-protein crosslinking in that cell line. Taken together, our results support the hypothesis that cadmium is not directly genotoxic.