Light regulates the c-fos protooncogene in the suprachiasmatic nucleus (SCN), with increased expression after animals are exposed to light during the dark phase of a light-dark cycle or during the subjective night in constant darkness. To determine whether this phase-dependent activation of c-Fos persists in an acute in vitro preparation, we prepared horizontal slices of hamster ventral hypothalamus, electrically stimulated the still-attached optic nerves, recorded the resulting evoked potentials in the SCN, and examined c-Fos protein levels in the nucleus by immunohistochemistry. The number of SCN cells labeled for immunoreactive c-Fos was significantly increased in slices stimulated during projected night, but not during projected day, compared to matched, sham-stimulated control slices. These results imply that the phase-dependent mechanism that gates c-Fos photoinduction in vivo is intrinsic to SCN tissue, and they suggest that an in vitro slice preparation will provide a useful model for dissecting the responsible signal transduction elements.