This study investigated the effects of gender, academic performance and preschool education on mental rotation skills among Turkish prospective teachers. A total of 525 undergraduate students (364 female) from a government university located in western Turkey completed the Mental Rotation Test (MRT). A three-way [2 (gender) × 5 (academic performance) × 2 (preschool education)] between-subjects analysis of variance (ANOVA) revealed that males outperformed females in mental rotation performance (d = 0.86). Significant effects of preschool education and academic performance on MRT scores were also observed. Significant interactions were observed between gender and academic performance and academic performance and preschool education. There was also a significant three-way interaction of gender, academic performance and preschool education. An interaction between gender and preschool education failed to reach significance.