This study aimed to determine the implementation of gender-segregated classrooms, students' learning enthusiasm, and the effect of classroom segregation between male and female students on students' learning enthusiasm in Islamic Religious Education (PAI) at SMK At-Taqwa Muhammadiyah Miri in the 2025/2026 academic year. This research employed a quantitative correlational approach. The population and sample consisted of 37 students. Data were collected through questionnaires, documentation, and observation. Data analysis included descriptive statistics, validity test, reliability test, normality test, linearity test, Product Moment correlation, simple linear regression, t-test, and coefficient of determination. The results showed that the implementation of classroom segregation was categorized as good with a mean score of 83.68, while students' learning enthusiasm was also categorized as good with a mean score of 82.49. The Product Moment correlation test yielded a correlation coefficient of 0.529 with a significance value of 0.001 < 0.05, indicating a positive and significant relationship between classroom segregation and students' learning enthusiasm. The simple linear regression analysis produced the equation Y = 41.618 + 0.488X, while the t-test result showed a t-value of 3.691 with a significance level of 0.001 < 0.05. The coefficient of determination (R²) was 0.280, indicating that classroom segregation contributed 28% to students' learning enthusiasm. Therefore, classroom segregation between male and female students has a positive and significant effect on students' learning enthusiasm in Islamic Religious Education at SMK At-Taqwa Muhammadiyah Miri.