Male and female organisms may have significant differences in their exposure, toxicokinetics, and response to chemicals, but gender effects have received relatively little attention, often viewed as a confounder rather than of primary importance. In this paper, we examine some of the key issues and methodologies for incorporating gender in studies of the effects of chemicals on wildlife, and explore bioindicators and biomarkers of gender effects. Examining gender-related differences in response to chemicals is complicated in wildlife because of the vast array of species, and differences in niches, lifespans, reproductive cycles and modes, and population dynamics. Further, organisms are more at risk in some ecosystems than others, which may increase the magnitude of effects. Only by studying wild animals, especially native species, can we truly understand the potential impact of gender-specific effects of chemical exposure on populations. Several factors affect gender-related differences in responses to chemicals, including exposure, age, size, seasonality, and genetic and phenotypic variation. There are clear examples where gender-related differences have had significant effects on reproductive success and population stability, including destabilization of gamete release in invertebrates, and alterations of endocrine and neuroendocrine system functioning in vertebrates. A wide range of new technologies and methods are available for examining gender-related differences in responses to chemicals. We provide examples that show that there are gender-related differences in responses to chemicals that have significant biological effects, and these gender-related differences should be taken into account by scientists, regulators, and policy makers, as well as the public.