Is there a direct retina-raphe-suprachiasmatic nucleus pathway in the rat?

Possible pathways from the retina to the suprachiasmatic nucleus (SCN) relaying in the dorsal raphe nucleus (DRN) were investigated in rats using combined anterograde and retrograde tracing with immunohistochemistry. After injection of wheat germ agglutinin-conjugated horseradish peroxidase-colloidal gold complex into the SCN, many neurons were retrogradely labeled in the middle levels of the DRN. Approximately one half of these neurons contained serotonin. After injection of cholera toxin B subunit into the eyes, a few anterogradely labeled afferent fibers were detected in the rostral DRN, however, not in contact with retrogradely labeled neurons. Our findings provide direct evidence that serotonergic projections to the rat SCN stem from the DRN nuclei. They also suggest that retina-raphe-SCN projections, a presumed third visual input to the mammalian circadian pacemaker, may include further neuronal connections or brain sites.