Developmental changes in the release of LHRH-1 (top) and kisspeptin-54 (bottom) in push-pull perfusates. Both peptides were measured in the same samples. Kisspeptin-54 levels gradually increased along with the pubertal increase in LHRH-1 release. The nocturnal increase in kisspeptin-54 release was already observed in prepubertal monkeys and continued through the pubertal period. In contrast, the nocturnal increase in LHRH-1 release occurred in early and midpubertal monkeys but not in prepubertal monkeys. The number of animals at the prepubertal, early pubertal, and midpubertal stages was six, six, and five, respectively. White bars, morning values; black bars, evening values. *, P < 0.05 vs. prepubertal; ***, P < 0.001 vs. prepubertal; +, P < 0.05 vs. early pubertal; ++, P < 0.01 vs. early pubertal; +++, P < 0.001 vs. early pubertal; a, P < 0.05 vs. morning; aa, P < 0.01 vs. morning; aaa, P < 0.001 vs. morning.