Purpose: The purpose of the current investigation was to retrospectively assess possible differences in physiological performance characteristics between junior cyclists signing a contract with an under-23 (U23) development team versus those failing to sign such a contract. Methods: Twenty-five male junior cyclists (age: 18.1 [0.7] y, stature: 181.9 [6.0] cm, body mass: 69.1 [7.9] kg, peak oxygen uptake: 71.3 [6.2] mL·min−1·kg−1) were assigned to this investigation. Between September and October of the last year in the junior category, each cyclist performed a ramp incremental exercise test to determine certain physiological performance characteristics. Subsequently, participants were divided in 2 groups: (1) those signing a contract with a U23 development team (JUNIORU23) and (2) those failing to sign such a contract (JUNIORNON-U23). Unpaired t tests were used to assess possible between-groups differences in physiological performance characteristics. The level of statistical significance was set at P < .05 two tailed. Results: No significant between-groups differences in submaximal (ie, gas exchange threshold, respiratory compensation point) and maximal physiological performance characteristics (ie, peak work rate, peak oxygen uptake) expressed in absolute values (ie, L·min−1, W) were observed (P > .05). However, significant between-groups differences were observed when physiological performance characteristics were expressed relative to the cyclists’ body weights (P < .05). Conclusions: The current investigation showed that junior cyclists stepping up to a U23 development team might be retrospectively differentiated from junior cyclists not stepping up based on certain physiological performance characteristics, which might inform practitioners and/or federations working with young cyclists during the long-term athletic development process.