Serotonin and substance P colocalization in medullary projections to the nucleus tractus solitarius: dual-colour immunohistochemistry combined with retrograde tracing.

Serotonin (5HT) and substance P (SP) are colocalized in terminals within the nucleus tractus solitarius (NTS). The purpose of the present study was to determine the origin of these terminals. 5HT- and SP-immunoreactivities (IR) were visualized using dual-colour immunofluorescence histochemistry with amino-4-methylcoumarin-3-acetic acid- and fluorescein isothiocyanate-conjugated secondary antisera, while NTS-afferent neurons were visualized by retrograde labelling with rhodamine beads. Extensive colocalization of 5HT- and SP-IR was seen in NTS-afferent neurons located in the nucleus raphe pallidus, nucleus raphe obscurus, nucleus raphe magnus, and in the parapyramidal region. Over 80 per cent of the SP-IR NTS-afferent neurons contained 5HT-IR, while 68 per cent of the 5HT-IR neurons contained SP-IR. Thus, 5HT- and SP-IR are extensively colocalized in NTS-afferent neurons in the medullary raphe nuclei and associated areas of the ventral medulla.