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Objectives: To investigate potential effects of heading on the neurocognitive performance and the white matter (WM) of the brain in high-level adult male football players., Design: Prospective longitudinal., Methods: Football players engaging in the highest football leagues in Germany were included. Neurocognitive performance tests and diffusion tensor imaging (DTI) were executed before and after the observation period. Video recordings of each training session and each match play during the observation period were analyzed regarding heading exposure and characteristics. Four DTI measures from tract-based spatial statistics (fractional anisotropy, mean, axial, and radial diffusivity) were investigated. Associations between heading variables and DTI and neurocognitive parameters were tested subsequently., Results: 8052 headers of 22 players (19.9 ± 2.7 years) were documented in a median of 16.9 months. The individual total heading number ranged from 57 to 943 (median: 320.5). Header characteristics differed between training sessions and matches. Neurocognitive performance (n = 22) and DTI measures (n = 14) showed no significant differences from pre- to post-test. After correction for multiple comparisons, no significant correlations with the total heading number were found. However, the change in fractional anisotropy in the splenium of the corpus callosum correlated significantly with the total amount of long-distance headers (Pearson's r = -0.884; p < 0.0001)., Conclusions: Over the median observation period of 16.9 months, DTI measures and neurocognitive performance remained unchanged. To elucidate the meaning of the association between individual change in fractional anisotropy and long-distance headers further investigations with larger samples, longer observations, and various cohorts regarding age and level of play are required., Competing Interests: Declaration of interest statement CR receives scientific funding for projects on traumatic brain injuries and heading from the Federal Institute of Sports Sciences (Germany) and the Heinz Nixdorf Westfalian Foundation. He is a member of the medical committee of the German Football association (DFB) and provides counseling on the management of traumatic brain injury to the Union of European Football Associations (UEFA). All other authors declare no conflicting interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)