This study investigates the release characteristics of neuropeptide Y (NPY) from young (10 weeks) and old (22 months) rat atrium. Levels of NPY release from samples of atrium were studied by organ perifusion. Rats were exposed to light:dark (LD) cycles of 12:12 or 18:6 and sacrificed at different circadian stages: 0, 4, 7, 12, 18, and 20 h after dark onset (HADO) for LD 12:12 or 0, 2, 3.5, 6, 15, and 22 HADO for LD 18:6. The heart was collected, and the right atrium was removed, weighed, and perifused with Krebs-bicarbonate buffer for 100 min, including a period of 50 min for stabilization of secretion rate. NPY concentrations released by atrium did not differ between the two age groups. NPY exhibited daily variations in concentrations in LD 12:12, with a peak during the end of scotophase, at 12 HADO, in both the young and old rats. These variations were strongly modified in LD 18:6, where the pattern of the release exhibited two peaks occurring during the two thirds of dark (3.5 HADO) and light (22 HADO) periods. This strongly suggests that the NPY rhythm is dependent on the environmental light:dark cycle. In this paper we show that NPY concentrations in the rat atrium exhibit daily variations, which are maintained with ageing. Moreover, photoperiod greatly influences NPY levels in the atrium.