Efferent projections of the suprachiasmatic nucleus: II. Studies using retrograde transport of fluorescent dyes and simultaneous peptide immunohistochemistry in the rat

Abstract

In a previous study (Watts et al., '87) we reexamined the projections of the suprachiasmatic nucleus (SCh) with the PHA‐L method and found that they could be divided conveniently into six groups of fibers. By far the densest projection ends just dorsal to the SCh in a comma‐shaped region designated the “subparaventricular zone,” although some fibers continue on through the paraventricular nucleus of the hypothalamus to end in the overlying midline thalamus, and others continue on to end in the dorsomedial nucleus, the region around the ventromedial nucleus, and the posterior hypothalamic area. Other relatively sparse projections from the SCh were also described to the preoptic region, lateral septal nucleus, parataenial and paraventricular nuclei of the thalamus, and ventral lateral geniculate nucleus. In addition, the same method was used to show that the subparaventricular zone projects in turn massively to these same regions, as well as back to the SCh itself and to the periaqueductal gray. The present series of experiments was designed to confirm these observations with retrograde tracer injections and to investigate the cellular and possible neurotransmitter organization of the major projections from the SCh and subparaventricular zone with a combined retrograde tracer‐immunohistochemical method. For this, the distribution of neuronal cell bodies within the SCh that stain with antisera to vasopressin, vasoactive intestinal polypeptide (VIP), corticotropin‐releasing factor, bombesin, substance P, neurotensin, somatostatin, thyrotropin‐releasing hormone, and angiotensin II was described in detail first. Then the distribution of retrogradely labeled neurons that were also stained for one or another of these peptides was described after injections of true blue, or in some cases SITS, into the regions of the subparaventricular zone, the paraventricular and parataenial nuclei of the thalamus, the ventromedial nucleus, the dorsomedial nucleus, and the periaqueductal gray. The results confirm previous immunohistochemical and anterograde tracing studies and in addition indicate that cells in dorsal as well as ventral parts of the SCh project to each of the terminal fields examined, as do many cells in surrounding areas, including the subparaventricular zone. Our results also suggest that, at the very least, vasopressin‐, VIP‐, and neurotensin‐stained cells in the SCh project to the subparaventricular zone, midline thalamus, and dorsomedial nucleus, and that the vasopressin and VIP‐stained fiber systems are partially segregated at the level of the subparaventricular zone. Finally, we have shown that neuropeptide‐Y‐stained neurons in the ventral lateral geniculate nucleus (the intergeniculate leaflet) project to the same nucleus on the opposite side of the brain.