The effects of light on the circadian rhythm in melatonin release from the pineal organ of a teleost, ayu (Plecoglossus altivelis) were investigated in flow-through culture. Under the reversed light-dark (LD) cycle, the melatonin rhythm phase shifted as compared with those under the normal LD cycle. This phase shift persisted even under constant darkness (DD). Single 6-h light pulses starting at six different circadian phases under DD acutely suppressed melatonin release. Phase-dependent phase shifts in the circadian rhythm of melatonin release were also observed. The phase response curve to light pulses in the ayu pineal organ is typical of that found in many circadian systems. Thus, the ayu pineal organ should provide a useful model for analyzing the physiological and molecular basis of the entrainment mechanism of vertebrate circadian system.