1.Isolation and cDNA cloning of a novel galanin-like peptide (GALP) from porcine hypothalamus.
Ohtaki T1, Kumano S, Ishibashi Y, Ogi K, Matsui H, Harada M, Kitada C, Kurokawa T, Onda H, Fujino M. J Biol Chem. 1999 Dec 24;274(52):37041-5.
Galanin is a widely distributed neuropeptide with a variety of physiological functions. Three galanin receptor subtypes, GALR1, GALR2, and GALR3, have been reported. We isolated a novel galanin-like peptide (GALP) from porcine hypothalamus by observing its activity for increasing [(35)S]GTPgammaS binding to a membrane preparation of GALR2-transfected cells. The peptide had 60 amino acid residues and a non-amidated C terminus. The amino acid sequence of GALP-(9-21) was completely identical to that of galanin-(1-13). A cloned porcine GALP cDNA indicated that GALP was processed from a 120-amino acid GALP precursor protein. The structures of rat and human GALP-(1-60) were deduced from cloned cDNA, which indicated that the amino acid sequences 1-24 and 41-53 were highly conserved between humans, rats, and pigs. Receptor binding studies revealed that porcine GALP-(1-60) had a high affinity for the GALR2 receptor (IC(50) = 0.24 nM) and a lower affinity for the GALR1 receptor (IC(50) = 4.
2.Distribution and chromatographic analysis of galanin message-associated peptide (GMAP)-like immunoreactivity in the rat.
Xu Y1, Rökaeus A, Johansson O. Regul Pept. 1994 Apr 14;51(1):1-16.
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.
3.Pharmacological and functional characterization of galanin-like peptide fragments as potent galanin receptor agonists.
Lang R1, Berger A, Santic R, Geisberger R, Hermann A, Herzog H, Kofler B. Neuropeptides. 2005 Jun;39(3):179-84. Epub 2005 Feb 1.
The hypothalamic galanin-like peptide (GALP) was isolated by its ability to activate galanin receptors. The mature porcine GALP is a 60-amino acid neuropeptide proteolytically processed from a 120-amino acid precursor protein. It contains a region identical to the N-terminal 13-amino acids of the neuropeptide galanin. Within the sequence of human GALP (1-60) a potential proteolytic cleavage site between two basic amino acids is present at position 33, which might lead to a shorter C-terminally amidated peptide. In addition, the first two amino acids could be potentially removed via the action of dipeptidase IV. Ligand binding assays using the human neuroblastoma cell line SH-SY5Y transfected with the respective galanin receptors revealed that human GALP (1-60) displayed the highest affinity for the galanin receptor subtype GalR3 (IC50 = 10 nM) followed by GalR2 (IC50 = 28 nM) and GalR1 (IC50 = 77 nM). Ligand binding assays and functional studies showed that the human GALP (3-32) fragment was at least as potent as full length GALP (1-60).
4.Distribution, regulation and role of hypothalamic galanin systems: renewed interest in a pleiotropic peptide family.
Gundlach AL1, Burazin TC, Larm JA. Clin Exp Pharmacol Physiol. 2001 Jan-Feb;28(1-2):100-5.
1. Galanin peptide and galanin receptor-binding sites are known to be widely distributed within the central nervous system, particularly in the hypothalamus in the preoptic area, the paraventricular (PVN) and supraoptic (SON) nuclei and the arcuate nucleus/median eminence. 2. The present brief review focuses on some recent studies of the regional and cellular localization of mRNA encoding galanin and two galanin receptor subtypes (GalR1 and GalR2) in the hypothalamus, regulation of galanin and/or galanin receptor expression in various nuclei by physiological stimuli, electrophysiological effects of galanin on hypothalamic neurons and the isolation and cloning of galanin-like peptide (GALP), a putative endogenous ligand for GalR2. 3. In situ hybridization studies in rat brain have demonstrated an abundance of GalR1 mRNA in SON, magnocellular (m) and parvocellular (p) PVN and dorsomedial, ventromedial and arcuate nuclei. In contrast, GalR2 mRNA is enriched in pPVN, but not mPVN, and is not detected in SON.