White adipose tissue is an important endocrine organ. Receptors for several hormones are found in the adipocytes, suggesting that these hormones may directly regulate the activity of the fat cells. The effects of prolactin (PRL), growth hormone (GH), melatonin, insulin and their interaction on the regulation of leptin secretion from ovine peri-renal adipose tissue samples were evaluated. Adipose tissue isolated from 15 ewes slaughtered in May, July (spring/summer), September, October and November (Fall) (3 ewes per month) were used. Adipose tissue was cut into 100mg samples, which were incubated for 2h in Eagle's medium or a medium supplemented with melatonin (100ng/ml) and challenged with or without (control) the following hormones: insulin (100ng/ml), PRL (100ng/ml), PRL (300ng/ml), insulin (100ng/ml)+PRL (100ng/ml), insulin (100ng/ml)+PRL (300ng/ml), GH (100ng/ml), or insulin (100ng/ml)+GH (100ng/ml). After the addition of the hormones, the incubation continued for another 3h period. Leptin concentrations in the culture media were determined using RIA. Results showed the basal secretion of leptin from adipose tissue to vary, depending on the month in which the incubation was carried out. Being the lowest in May (end of spring) and the highest during the fall months (P<0.05). However, no significant individual effects of the added hormones on leptin concentration were recorded. These results suggest that all the examined hormones may affect the activity of the adipocytes. However the hormones’ impacts depend on the presence of other hormones. Similarly, there was a substantial seasonality effect recorded regarding the interaction between the hormones.