Transforming growth factor-alpha (TGF-alpha) is abundantly expressed in the suprachiasmatic nucleus of several rodent species. It was recently suggested to be a clock output signal regulating the activity/rest rhythm. In this study we further characterized the cellular identity of TGF-alpha-expressing cells in the suprachiasmatic nucleus of the Syrian hamster (Mesocricetus auratus). Using confocal laser scanning fluorescence imaging on brain sections immuno-histologically processed for TGF-alpha and GFAP double staining, we observed that in the suprachiasmatic nucleus, TGF-alpha staining is located mainly in GFAP-positive cells, indicating suprachiasmatic nucleus astrocytes produce TGF-alpha. Glial expression of TGF-alpha was also observed in the 3rd ventricle tanycytes of the retrochiasmatic area. In other brain regions where the TGF-alpha message is abundant, such as in the piriform cortex, we observed that TGF-alpha staining is mainly located in neurons. Our results provide the first evidence that glial cells are involved in the regulation of output from the suprachiasmatic nucleus circadian clock through a diffusible mechanism.