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Molecular characterization and expression of cloned human galanin receptors GALR2 and GALR3

Academic Article
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Overview

authors

  • Kolakowski, L. F.
  • O'Neill, G. P.
  • Howard, A. D.
  • Broussard, S. R.
  • Sullivan, K. A.
  • Feighner, S. D.
  • Sawzdargo, M.
  • Nguyen, T.
  • Kargman, S.
  • Shiao, L. L.
  • Hreniuk, D. L.
  • Tan, C. P.
  • Evans, J.
  • Abramovitz, M.
  • Chateauneuf, A.
  • Coulombe, N.
  • Ng, G.
  • Johnson, M. P.
  • Tharian, A.
  • Khoshbouei, H.
  • George, S. R.
  • Smith, Roy
  • O'Dowd, B. F.

publication date

  • December 1998

journal

  • Journal of Neurochemistry  Journal

abstract

  • Galanin is a 29- or 30-amino acid peptide with wide-ranging effects on hormone release, feeding behavior, smooth muscle contractility, and somatosensory neuronal function. Three distinct galanin receptor (GALR) subtypes, designated GALR1, 2, and 3, have been cloned from the rat. We report here the cloning of the human GALR2 and GALR3 genes, an initial characterization of their pharmacology with respect to radioligand binding and signal transduction pathways, and a profile of their expression in brain and peripheral tissues. Human GALR2 and GALR3 show, respectively, 92 and 89% amino acid sequence identity with their rat homologues. Radioligand binding studies with 125I-galanin show that recombinant human GALR2 binds with high affinity to human galanin (K(D) = 0.3 nM). Human GALR3 binds galanin with less affinity (IC50 of 12 nM for porcine galanin and 75 nM for human galanin). Human GALR2 was shown to couple to phospholipase C and elevation of intracellular calcium levels as assessed by aequorin luminescence in HEK-293 cells and by Xenopus melanophore pigment aggregation and dispersion assays, in contrast to human GALR1 and human GALR3, which signal predominantly through inhibition of adenylate cyclase. GALR2 mRNA shows a wide distribution in the brain (mammillary nuclei, dentate gyrus, cingulate gyrus, and posterior hypothalamic, supraoptic, and arcuate nuclei), and restricted peripheral tissue distribution with highest mRNA levels detected in human small intestine. In comparison, whereas GALR3 mRNA was expressed in many areas of the rat brain, there was abundant expression in the primary olfactory cortex, olfactory tubercle, the islands of Calleja, the hippocampal CA regions of Ammon's horn, and the dentate gyrus. GALR3 mRNA was highly expressed in human testis and was detectable in adrenal gland and pancreas. The genes for human GALR2 and 3 were localized to chromosomes 17q25 and 22q12.2-13.1, respectively.

subject areas

  • Amino Acid Sequence
  • Animals
  • Blotting, Northern
  • Brain
  • Cell Line
  • Cloning, Molecular
  • Humans
  • Isomerism
  • Ligands
  • Mice
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Rats
  • Receptors, Galanin
  • Receptors, Neuropeptide
  • Ribonucleases
  • Signal Transduction
  • Swine
  • Xenopus laevis
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Research

keywords

  • G protein-coupled receptor
  • cloning
  • galanin receptor
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Identity

International Standard Serial Number (ISSN)

  • 0022-3042

Digital Object Identifier (DOI)

  • 10.1046/j.1471-4159.1998.71062239.x

PubMed ID

  • 9832121
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Additional Document Info

start page

  • 2239

end page

  • 2251

volume

  • 71

issue

  • 6

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