ABSTRACT: Neck pain is a common musculoskeletal problem often accompanied by reduced exercise-induced hypoalgesia (EIH) or hyperalgesia compared to an asymptomatic population. This study investigated EIH in a healthy population during experimental neck pain. Forty participants were randomized into this double-blinded parallel-group study. On four separate test days (Day0, Day2, Day4, Day15), participants completed the Neck Disability Index (NDI) and scored neck pain intensity during head movements on a numerical rating scale (NRS). At the end of Day0 and Day2, Nerve Growth Factor (NGF) or isotonic saline (control) was injected into the right splenius capitis muscle. Pressure pain thresholds (PPTs) were recorded bilaterally over splenius capitis (neck), temporalis (head) and tibialis anterior (leg) muscles on all days. On Day0, Day4 and Day15, PPTs were recorded before and after a hand-bike exercise. EIH was defined as the PPT increase caused by the exercise. Compared with the control-group, the NGF-group demonstrated higher NDI scores at Day2 and Day4 (P<0.001,η2>0.557) and higher NRS scores (P<0.03,η2>0.09) along with reduced neck PPTs (P<0.01,d>0.44) at Day2(Right:95%CI[26.0,54.0];Left:95%CI[6.8,26.9]), Day4(Right:95%CI[40.5, 67.9];Left:95%CI[6.9,28.2]) and Day15(Right:95%CI[5.6,37.2];Left:95%CI[6.9,34.8]). Across days, the EIH-effect was reduced at the neck site in the NGF-group compared to the control-group (P<0.001,η2P=0.367,95%CI[-34.5,-13.7]). At the head and leg sites, the NGF-group showed reduced EIH-effect compared to the control-group (P<0.05,d>0.43) on Day4(Head:95%CI[-61.4,-22.9];Leg:95%CI[-154.7,-72.4]) and Day15(Head:95%CI[-54.3,-7.6];Leg:95%CI[-122.7,-34.4]). These results indicate that a few days of clinically comparable neck pain and hyperalgesia might have a negative impact on EIH-responses and may help explain why some neck pain patients do not experience immediate positive effects of exercise.