Anthropogenic disturbances cause the environment to change relatively fast. It is reasonable to assume that it is very unlikely for individuals to develop genetic adaptations to their polluted habitats, since adaptation through natural selection is a relatively slow process. Nevertheless, several studies have shown that such adaptations to changing environmental conditions may develop faster than anticipated. This study investigates the impact of historical metal pollution on a natural population of the gudgeon, Gobio gobio. Specimens from a contaminated site and a reference population were subjected to a series of three exposure experiments to cadmium after an acclimation period to reconstituted fresh water of 36 days. First, we performed an acute toxicity test on a sub-sample of both experimental groups to determine times-to-death (TTD) and lethal body burdens (LBB). The remaining individuals were used in a chronic Cd-exposure experiment, after which total Cd-body concentration, as well as Cd-concentrations and metallothionein-like protein (MTLP) levels in liver and gill tissue were determined. From the specimens that were not sacrificed for these measurements, a random subsample was subjected to a second acute toxicity test to evaluate the effect of chronic Cd-exposure (acclimation) on TTD and LBB. Our results show that, particularly after an extra acclimation period to a sublethal Cd-concentration, specimens originating from the contaminated sample area survived the acute exposure experiments better, despite the fact that neither the average Cd-accumulation rate, nor the lethal body concentrations differed between fishes from both groups. We also find that gudgeons from both populations translocated Cd from the gills (and probably also from other compartments) to the liver, where it can be more efficiently detoxified by MTLPs. Indeed, MTLP levels were found to increase faster in liver and gill tissue of specimens from the contaminated site, resulting in significantly higher MTLP-levels in the organs of these fishes. Although this study does not provide direct evidence for a genetic basis of Cd resistance (i.e. at the gene level), our results indicate that the regulation of MTLP-gene expression may involve a genetic component.