The effects of photoperiod on the development of central neurotransmitters were investigated with tilapia, Oreochromis mossambicus. Zero-day-old (the hatching day) tilapia were raised in three different photoperiods (light/dark cycle): 12/1, 24/0, and 0/24 h, respectively. On the 5th day, brain serotonin (5-HT), norepinephrine (NE), γ-aminobutyric acid (GABA), and glutamate (Glu) contents were quantified by a high-performance liquid chromatograph with electrochemical detection. Similar experiments were performed on the 5-, 10-, 15-, 20-, and 25-day-olds. These results showed that the photoperiod influenced both brain NE and GABA contents during its respective restricted period, before days 10 posthatching. Brain 5-HT content was influenced, either facilitated or suppressed according to the developing stage, whereas, brain Glu content was not altered by the different photoperiod exposure throughout the present studies.