Selective anterograde tracing of the individual serotonergic and nonserotonergic components of the dorsal raphe nucleus projection to the vestibular nuclei

It is well known that the dorsal raphe nucleus (DRN) sends serotonergic and nonserotonergic projections to target regions in the brain stem and forebrain, including the vestibular nuclei. Although retrograde tracing studies have reported consistently that there are differences in the relative innervation of different target regions by serotonergic and nonserotonergic DRN neurons, the relative termination patterns of these two projections have not been compared using anterograde tracing methods. The object of the present investigation was to trace anterogradely the individual serotonergic and nonserotonergic components of the projection from DRN to the vestibular nuclei in rats. To trace nonserotonergic DRN projections, animals were pretreated with the serotonergic neurotoxin 5,7-dihydroxytryptamine (5,7-DHT), and then, after 7 days, the anterograde tracer biotinylated dextran amine (BDA) was iontophoretically injected into the DRN. In animals treated with 5,7-DHT, nonserotonergic BDA-labeled fibers were found to descend exclusively within the ventricular plexus and to terminate predominantly within the periventricular aspect of the vestibular nuclei. Serotonergic DRN projections were traced by injecting 5,7-DHT directly into DRN, and amino–cupric–silver staining was used to visualize the resulting pattern of terminal degeneration. Eighteen hours after microinjection of 5,7-DHT into the DRN, fine-caliber degenerating serotonergic terminals were found within the region of the medial vestibular nucleus (MVN) that borders the fourth ventricle, and a mixture of fine- and heavier-caliber degenerating serotonergic terminals was located further laterally within the vestibular nuclear complex. These findings indicate that fine-caliber projections from serotonergic and nonserotonergic DRN neurons primarily innervate the periventricular regions of MVN, whereas heavier-caliber projections from serotonergic DRN neurons innervate terminal fields located in more lateral regions of the vestibular nuclei. Thus, serotonergic and nonserotonergic DRN axons target distinct but partially overlapping terminal fields within the vestibular nuclear complex, raising the possibility that these two DRN projection systems are organized in a manner that permits regionally-specialized regulation of processing within the vestibular nuclei.