Objectives: To investigate the relationship between mental health issues and medical career interest among undergraduate premedical students, and to explore whether this association varies by gender. Methods: A longitudinal survey of freshman and sophomore premedical students at Indiana University was conducted during the 2015-16 academic year. Survey data were collected from 390 respondents via an online questionnaire (response rate=14%) in September 2015, and 292 of these respondents participated in the follow-up survey in April 2016 (retention rate=75%). Multi-level regression models were used to estimate the associations among depressive symptoms, burnout, and medical career interest. Results: Respondents who experienced more depressive symptoms (beta = -.07, z =-2.49, p =.013) and higher levels of burnout (beta = -.50, z =-3.98, p <.001) reported significant reductions in medical career interest over the study period. These associations remained consistent after controlling for socio-demographic characteristics and prior academic achievement. Depressive symptoms were associated with steeper declines in medical career interest among women compared to men (beta = -.09, z =-2.01, p =.045), though the relationship between burnout and medical career interest did not significantly vary by gender. Conclusions: Results provide evidence that premeds who experience more depressive symptoms and higher levels of burnout become less interested in entering the medical profession. The negative association between depressive symptoms and medical career interest was even more pronounced among premedical women compared to men. Findings suggest that colleges and universities in the United States should implement programs aimed at safeguarding the mental health of premedical students.