We aimed to examine the acute effect of mental fatigue on young basketball players’ three-dimensional multiple object tracking (3D-MOT) skills. Our participants were 12 adolescent basketball players ( M age = 16.66, SD = 1.87 years; M years of practice = 2.66, SD = 1.07 years). In nine lab visits, we used visits 1 to 7 to familiarize participants with 3D-MOT, a subjective scale of mental fatigue, and a Stroop task involving mental set shifting. In the last two visits, participants performed in both experimental (EXP) and control (CON) conditions that were presented in randomized order. In the EXP condition, participants performed 3D-MOT pre- and post-60 minutes of induced mental fatigue; in the CON condition, they watched a documentary. After each condition, B participants performed the National Aeronautics and Space Administration Task Load Index (NASA-TLX). 3D-MOT performance measures were the “score” and “fastest trial score success.” The response time on the Stroop tasks increased throughout the mental fatigue inducement in the experimental condition ( p = .0037). The NASA-TLX responses were higher following the EXP condition than following the CON condition for mental demand, temporal demand, and performance (all ps < .05). Still, there were no significant EXP versus CON differences on the 3D-MOT performance indicators.