Introduction: In Florida, a state that consistently leads the nation in adverse sexual health outcomes among adolescents, numerous demographic and socio-cultural differences exist across the North, Central, and South regions. However, little is known about regional differences in sexuality education and beliefs among teachers. Methodology: Using a mail-based survey of 462 teachers, we examined regional differences in parental consent requirements, abstinence instruction, student language barriers, availability of Spanish curricula, teacher beliefs regarding the dissemination of accurate safer sex information, and missing data. t-tests and multivariate logistic regression analyses were conducted. Results: North Floridian teachers taught more abstinence than those in Central and South Florida. Additionally, they had fewer students with language barriers, offered less Spanish curricula, and, in general, possessed less favorable attitudes toward presenting accurate safer sex information. However, South Floridian teachers were most likely to report parental consent requirements in their schools. These findings largely remained in multivariate analyses. Conclusions: The present study identified several noteworthy regional differences in sexuality education in Florida. Recommendations are given for developing and implementing programs that account for these differences within the state. (Contains 3 tables.)