In zebrafish, pineal arylalkylamine-N-acetyltransferase (zfaanat2) mRNA expression begins at 22 h post-fertilization (hpf), and the clock-controlled rhythm of its transcript begins on the third day of development. Here we describe the role of light and of the clock gene, period2 (zper2) in the development of this rhythm. In 1-day-old zebrafish embryos, zper2 expression is transiently up-regulated by light in the pineal gland and, to a lesser extent, in other areas of the brain. Expression of zper2 that was not affected by light occurred in the olfactory placode and lactotroph cells of the pituitary primordium. Circadian analysis of pineal zfaanat2 mRNA expression indicated that light exposure is required for proper development of the circadian clock-controlled rhythmic expression of this gene. Knockdown of zPER2 using antisense technology abolished the effect of light on development of the zfaanat2 rhythm in the pineal gland, corroborating the role of zper2 in light entrainment of the circadian oscillator in zebrafish. Further analysis of zper2 expression at earlier stages of development revealed that light exposure at the blastula to mid-segmentation stages also caused a transient increase in zper2 expression. At mid-segmentation, before pineal differentiation, light-induced zper2 expression was enhanced in pineal progenitor cells. Thus, a possible role for early photoreception and light-induced zper2 expression in the development of clock-controlled rhythms remains to be investigated.