Immunohistochemical evidence of substance P-like immunoreactivity in some 5-hydroxytryptamine-containing neurons in the rat central nervous system

Abstract With the indirect immunofluorescence technique of Coons and collaborators a possible coexistence of 5-hydroxytryptamine (5-HT) and substance P in neurons of the lower medulla oblongata was explored. Antisera to 5-HT and to dopadecarboxylase (aromatic l -aminoacid decarboxylase), an enzyme probably present in immunologically indistinguishable forms both in catecholamine and 5-HT neurons, were used as markers for 5-HT neurons and an antiserum raised to synthetic substance P conjugated with bovine serum albumin for substance P-containing neurons. Five or 10 μm thick, consecutive sections were stained with the three antisera. Numerous cell somata in nucleus raphe magnus, nucleus raphe obscurus, nucleus raphe pallidus, pars α of the nucleus reticularis gigantocellularis and nucleus interfascicularis hypoglossi contained both substance P-like immunoreactivity and 5-HT (and dopadecarboxylase) immunoreactive material. After intraventricular or intracisternal injections of 5,6- or 5,7-dihydroxytryptamine, two neurotoxins assumed to cause degeneration mainly of 5-HT neurons, enlarged substance P and 5-HT (and dopadecarboxylase) positive fibres were seen in, around and lateral to the olivary complex. Furthermore, in these rats both substance P and 5-HT positive nerve terminals in the ventral horns of the spinal cord disappeared. These findings indicate that substance P and 5-HT may coexist not only in some cell bodies but also in axons and nerve endings. The latter conclusion must, however, remain tentative since the neurotoxins may cause unspecific damage and thus not only damage 5-HT (and postulated ‘SP-5-HT’) neurons. In further experiments reserpine was used, a drug known to deplete monoamines by affecting their storage sites. With a high dose of reserpine a marked depletion of 5-HT was obtained both in nerve terminals and cell bodies whereas substance P immunoreactive material seemed unaffected. Possible interpretations of these findings is that substance P and 5-HT have different storage sites within the neuron, or that reserpine selectively causes loss of 5-HT and not substance P from the same storage site.

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