||The prevalence of overtraining (OT) may vary anywhere from 10-60%, depending on the sport and athletic level. OT poses many problems to coaches, teams, and athletes as it leads to performance decrements, injury, and lost playing time. The physiological development of OT is not well understood and coaches lack affordable and easy-to-use tools that can be used for the early detection of OT, aside from psychometric tests. The purpose of this pilot study was to develop a protocol that could be used to examine the early psychophysiological development of OT, or functional overreaching (FOR) in order to better identify tools for the early detection of OT. Ten age-group cyclists completed 3 40k cycling time trials (TT) on 3 consecutive days with a 48-hour follow-up period to try to elicit a FOR response. Five measurements were used in this study to examine neuromuscular fatigue, changes in mood, affect, and perceived muscle pain and fatigue. Despite signs of acute fatigue, there were no changes in performance (p > 0.582) or in neuromuscular fatigue (p > 0.360) from one TT to the next. There was, however, a significant decrease in heart rate (p < 0.005) and changes in mood and affect indicative of some cumulative fatigue. The Fatigue subscale of the Profile of Mood States increased and remained elevated across three pre-TT time points, and positive affect (Positive and Negative Affect Schedule) decreased across two time points (p < 0.05 and 0.02, respectively). While these results do not support the conclusion that FOR was achieved in these age-group cyclists according to the definition provided by Meeusen et al., the significant increases in cumulative fatigue suggest that psychological stress and fatigue may be preliminary signs to even the first and earliest stage of OT, FOR. Therefore, future research should expand on the current study in order to develop a protocol that elicits a FOR response in the majority of subjects.