Female mate choice preferences may be the result of innate factors, experience, or an interaction between the two mechanisms. Understanding the relative contribution and possible interaction between these mechanisms is important for identifying sources of variation in behaviors under sexual selection. In this study, we evaluate the contribution of early experience on the development of mate choice preferences in the Neotropical Túngara Frog, Physalaemus pustulosus. We reared frogs from tadpole stage through sexual maturity in four acoustic treatments: the first group heard a conspecific chorus of Túngara Frogs, the second group heard a heterospecific chorus of the closely related sympatric congener P. enesefae, the third group heard no frog sounds, and the fourth group heard only broadband white noise. At sexual maturity, we tested each female's preferences for conspecific complex vs simple calls and discrimination against calls of the sympatric congener. Female choices in all of these tests were consistent with those in previous studies of wild-caught and laboratory-reared specimens of this species. The acoustic rearing environments in this study did not alter the preferences of females for complex conspecific calls or the discrimination of females against the sympatric congener. This study supports the hypothesis that early experience does not alter the mate choice preferences of female P. pustulosus.