The extent to which characteristic adolescent behaviors are associated with pubertal changes or driven by more general, puberty-independent developmental alterations is largely unknown. Using physiological and hormonal markers of puberty, this experiment characterized pubertal timing across adolescence and examined the relationships among these variables and novelty-directed behaviors. Males and females were tested for response to novelty at P28, P32, P36, P40, P44, P48, and P75, and examined for balano-preputial skinfold separation and sperm presence (males) or vaginal opening (females), followed by blood collection for hormonal assessments. Despite earlier pubertal maturation in females, with maturation generally completed by P36 in females and P44 in males, novelty-directed behavior peaked at P32 and P36 in both sexes, and was unrelated to pubertal measures. These data support the suggestion that the ontogenetic peak in this behavior during adolescence is not notably puberty dependent. © 2011 Wiley Periodicals,Inc. Dev Psychobiol 54: 523–535, 2012.