The present study investigated the spatial ability of adolescent athletes with different sport expertise of axial rotation, providing the cognitive characteristics from both behavior and functional near-infrared spectroscopy (fNIRS) tests. 16 basketball and soccer players, 15 freestyle athletes and 15 runners were selected for spatial ability testing with a three-factor mixed experimental design of 3 (sport type: open high-spatial sport, closed high-spatial sport, closed low-spatial sport) × 2 (rotation angle: 45°, 90°) × 3 (rotation axis: horizontal, vertical, longitudinal). Repeated-measures ANOVA was performed to evaluate the behavioral and fNIRS data of the mental rotation test. The behavioral data showed that the closed high-spatial (CH) group showed better performance than the open high-spatial (OH) and closed low-spatial (CL) group for large angle for every rotation axes. The fNIRS results showed the CH group had greater brain activation than the OH and CL group in the left dorsolateral superior frontal gyrus, precentral gyrus, postcentral gyrus, superior parietal gyrus, left angular gyrus, left precuneus and middle occipital gyrus. The findings provide additional empirical support for relating body rotation experience to spatial cognition performance.