Distribution and chromatographic analysis of galanin message-associated peptide (GMAP)-like immunoreactivity in the rat

Abstract

Galanin message-associated peptide (GMAP) constitutes the C-terminal part of the precursor protein encoding also the biologically active neuropeptide galanin (GAL). We have raised antisera against a species-conserved portion of GMAP, and investigated the localization of GMAP-like immunoreactivity (-LI) in relation to that of GAL-LI in the rat central and peripheral nervous system using the indirect immunofluorescence technique. In the central nervous system, GMAP-immunoreactive (-IR) cell bodies were observed in the hypothalamus, while GMAP-IR nerve fibers were demonstrated in the septum, hypothalamus, pons and spinal cord. In the posterior pituitary and in the connecting infundibular stalk, weakly fluorescent GMAP-IR nerve fibers were observed. GMAP-IR nerve fibers were also observed throughout the gastrointestinal tract, i.e., from the stomach down to the colon, and in all layers, except in the epithelium, of the wall. In general, the staining of consecutive tissue sections suggested that GMAP-IR was co-distributed with that of GAL-IR. A sensitive radioimmunoassay (RIA) for characterization of GMAP-IR in the rat central and peripheral nervous system was also developed. Characterization of GMAP-LI in acid extracts of rat brain and small intestine, using reverse phase high pressure liquid chromatography (rpHPLC), revealed multiple GMAP-IR forms that co-eluted with a synthetic porcine GMAP(19–41)-amide fragment, or were less or more polar than this fragment. The corresponding chromatographic analysis of GAL-LI revealed only one major form corresponding to rat GAL. The immunohistochemical data indicate that a GMAP-like peptide(s) probably is axonally transported and may possibly have pre- and/or post-synaptic functions. The nature of the multiple GMAP-IR components remains to be investigated, but may tentatively represent differently processed and/or chemically modified forms of rat GMAP(1–60).