In recent decades, pyrethroid pesticides have been deemed a safer alternative to previously used pesticides. While some evidence supports this assumption in mammals and birds, exposure to certain pyrethroids can affect concentrations of hormones vital to reproduction in fish. Thus, we hypothesized that pyrethroid exposure impacts fish reproductive behavior and the expression of genes associated with reproduction. We tested our hypothesis by examining effects of the widely used pyrethroid pesticide, bifenthrin, on the reproductive behaviors of the broadly distributed livebearing western mosquitofish, Gambusia affinis. We exposed sexually mature female fish to one of five environmentally relevant concentrations of bifenthrin and conducted behavioral assays to assess reproductive, social, and space use behaviors before and after exposure. We did not detect changes in behaviors measured in response to bifenthrin. However, exposure was associated with increased expression of an estrogen receptor gene (ER-α) and glucocorticoid receptor (GR) in brain tissue at bifenthrin concentrations at concentrations of 5.90 and 24.82 ng/L, and 5.90 and 12.21 ng/L, respectively. Our study supports the perspective that the use of multiple endpoints through integrative approaches is essential for understanding the cumulative impact of pollutants. Integrating physiological, morphological, and behavioral investigations of nonlethal concentrations of pollutants like bifenthrin may heighten our potential to predict their impact on individuals, populations, and communities.