Circadian rhythmicity was studied by following the course of N-acetyl-transferase activity in the pineal glands of chickens in vitro. The results indicate (i) a daily change during day 1 of organ culture in constant dark that was dependent on the time the chickens were killed, (ii) equivocal persistence of the daily change in constant dark during 6 to 7 days of organ culture, (iii) an effect of light, (iv) inhibition by adrenergic agents and cycloheximide, and (v) stimulation by dibutyryl adenosine 3',5'-monophosphate and related compounds.

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N-Acetyltransferase, an enzyme involved in melatonin production in the pineal gland, exhibits a circadian rhythm in chickens with peak values in the dark-time and low values during the light-time, commencing at lights-on. When pineal glands of chickens killed during the dark-time (with high N-acetyltransferase activity) were organ-cultured, there was a decline in enzyme activity to light-time values. Regardless of the time of the dark at which the chickens were killed, the enzyme activity reached light-time levels at precisely the same time.

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