Although hormones are vital to an organism’s ability to respond to environmental stressors, they can be directly altered by the environment and impact reproductive behavior. For example, in some fishes, aquatic hypoxia (low dissolved oxygen) inhibits the aromatase enzyme that converts testosterone to estradiol. Here, we examined the effects of short-term aromatase inhibition on reproductive behavior in male Pseudocrenilabrus multicolor, a widespread African cichlid, from one normoxic river population and one hypoxic swamp population. We further tested the response of females to treated and untreated males. We predicted that aromatase inhibition would decrease courtship and competitive behaviors, but the swamp population would be less affected given generational exposure to hypoxia. Specifically, we compared competition and courtship behavior of males treated with a short-term exposure to an aromatase inhibitor with control fish from the two populations. We found that both courtship and competitive behaviors were affected by the interaction between treatment and population. River fish performed fewer courtship and competitive behaviors under the aromatase inhibition treatment while the behavior of swamp males was unaffected. Additionally, we found that females from the swamp population preferred males from the aromatase inhibition treatment and river females preferred control males. While we found behavioral effects of short-term aromatase inhibition, we did not find any effects on male nuptial coloration. Overall, these results indicate that the effects of short-term aromatase inhibition on behavior could depend on local adaptation in response to hypoxia.