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Heading in soccer increases serum neurofilament light protein and SCAT3 symptom metrics. Objectives: To determine the effect of heading a soccer ball on serum neurofilament light (NF-L) protein, plasma tau protein and symptom metrics including total number of symptoms reported and symptom severity scores on the Standardized Concussion Assessment Tool- 3rd edition (SCAT3). Methods: Eleven male collegiate soccer players were recruited to take part in three experimental conditions including heading, sham and control conditions. Participants were required to perform 40 headers in 20 min in the heading condition, and control 40 soccer balls directed at them with their hands, chest or thigh in the sham condition. No ball contact was made during the control condition. Blood sampling and SCAT3 symptom assessments were completed prior to and 1 hour following conditions. A subset of participants returned 3 weeks following the heading condition for blood sampling. Results: NF-L was elevated at 1 hour (p=0.004) and 1 month (p=0.04) following the heading condition, and at 1 hour (p=0.02) following the control condition. Tau levels remained unchanged following all conditions. The total number of symptoms (TS) and symptom severity (SS) scores from the SCAT3 were both elevated following the heading condition (p=0.01 and p=0.03, respectively). Both TS and SS decreased following sham (p=0.04 and p=0.04) and control conditions (p=0.04 and p=0.04). Conclusion: An acute bout of soccer heading is associated with increased NF-L concentrations at 1 hour and 1 month following the session and can lead to symptoms commonly reported following sport-related concussion.