Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 379, Issue 4, pp 417–420 | Cite as

Analyzing the validity of GalR1 and GalR2 antibodies using knockout mice

Short Communication

Abstract

G-protein-coupled receptors (GPCRs) comprise the largest family of cell surface receptors and are the major drug targets for the treatment of various human diseases. The lack of sensitive and selective antibodies capable of recognizing endogenous GPCRs, however, hampers the progress of research on this class of receptors. GalR1 through GalR3, GPCRs for the neuropeptide galanin, are potential drug targets for seizure, Alzheimer’s disease, depression and anxiety, as well as pain and metabolic syndrome; therefore, determining the cellular and subcellular localization of galanin receptors is of high interest. Several Antibodies raised against galanin receptors are currently available from commercial or academic sources. We have tested several antibodies to GalR1 and GalR2 on tissues from respective knockout mice. Unexpectedly, the immunoreactivity patterns are the same in wild-type and in knockout mice, suggesting that current GalR1 and GalR2 antibodies, under standard immunodetection conditions, might not be suitable for mapping the receptors. These findings argue for taking precaution when using antibodies to galanin receptors.

Keywords

Galanin Antibody CNS 

Notes

Acknowledgments

We thank Jacinta Lucero (University of California, San Diego) for her help in testing various GalR antibodies. We thank Bruno Conti and Brendon Ross for reading the manuscript and providing helpful comments. The study was supported by MH074055 and MH063080 (T.B).

References

  1. Bailey KR, Pavlova MN, Rohde AD, Hohmann JG, Crawley JN (2007) Galanin receptor subtype 2 (GalR2) null mutant mice display an anxiogenic-like phenotype specific to the elevated plus-maze. Pharmacol Biochem Behav 86:8–20PubMedCrossRefGoogle Scholar
  2. Bartfai T, Hokfelt T, Langel U (1993) Galanin—a neuroendocrine peptide. Crit Rev Neurobiol 7:229–274PubMedGoogle Scholar
  3. Counts SE, Perez SE, Mufson EJ (2008) Galanin in Alzheimer’s disease: neuroinhibitory or neuroprotective? Cell Mol Life Sci 65:1842–1853PubMedCrossRefGoogle Scholar
  4. Crawley JN (2008) Galanin impairs cognitive abilities in rodents: relevance to Alzheimer’s disease. Cell Mol Life Sci 65:1836–1841PubMedCrossRefGoogle Scholar
  5. Elliott-Hunt CR, Pope RJ, Vanderplank P, Wynick D (2007) Activation of the galanin receptor 2 (GalR2) protects the hippocampus from neuronal damage. J Neurochem 100:780–789PubMedCrossRefGoogle Scholar
  6. Gottsch ML, Zeng H, Hohmann JG, Weinshenker D, Clifton DK, Steiner RA (2005) Phenotypic analysis of mice deficient in the type 2 galanin receptor (GALR2). Mol Cell Biol 25:4804–4811PubMedCrossRefGoogle Scholar
  7. Hawes JJ, Picciotto MR (2005) Erratum: characterization of GalR1, GalR2, and GalR3 immunoreactivity in catecholaminergic nuclei of the mouse brain. J Comp Neurol 490:98–100CrossRefGoogle Scholar
  8. He B, Counts SE, Perez SE, Hohmann JG, Koprich JB, Lipton JW, Steiner RA, Crawley JN, Mufson EJ (2005) Ectopic galanin expression and normal galanin receptor 2 and galanin receptor 3 mRNA levels in the forebrain of galanin transgenic mice. Neuroscience 133:371–380PubMedCrossRefGoogle Scholar
  9. Hohmann JG, Jureus A, Teklemichael DN, Matsumoto AM, Clifton DK, Steiner RA (2003) Distribution and regulation of galanin receptor 1 messenger RNA in the forebrain of wild type and galanin-transgenic mice. Neuroscience 117:105–117PubMedCrossRefGoogle Scholar
  10. Hokfelt T, Xu ZQ, Shi TJ, Holmberg K, Zhang X (1998) Galanin in ascending systems. Focus on coexistence with 5-hydroxytryptamine and noradrenaline. Ann N Y Acad Sci 863:252–263PubMedCrossRefGoogle Scholar
  11. Jacoby AS, Hort YJ, Constantinescu G, Shine J, Iismaa TP (2002) Critical role for GALR1 galanin receptor in galanin regulation of neuroendocrine function and seizure activity. Brain Res Mol Brain Res 107:195–200PubMedCrossRefGoogle Scholar
  12. Kuteeva E, Hokfelt T, Wardi T, Ogren SO (2008) Galanin, galanin receptor subtypes and depression-like behaviour. Cell Mol Life Sci 65:1854–1863PubMedCrossRefGoogle Scholar
  13. Larm JA, Shen PJ, Gundlach AL (2003) Differential galanin receptor-1 and galanin expression by 5-HT neurons in dorsal raphe nucleus of rat and mouse: evidence for species-dependent modulation of serotonin transmission. Eur J Neurosci 17:481–493PubMedCrossRefGoogle Scholar
  14. Leibowitz SF (1989) Hypothalamic neuropeptide Y, galanin, and amines. Concepts of coexistence in relation to feeding behavior. Ann N Y Acad Sci 575:221–233 discussion 233–225PubMedCrossRefGoogle Scholar
  15. Lerner JT, Sankar R, Mazarati AM (2008) Galanin and epilepsy. Cell Mol Life Sci 65:1864–1871PubMedCrossRefGoogle Scholar
  16. Lu X, Sharkey L, Bartfai T (2007) The brain galanin receptors: targets for novel antidepressant drugs. CNS Neurol Disord Drug Targets 6:183–192PubMedCrossRefGoogle Scholar
  17. Lu X, Ross B, Sanchez-Alavez M, Zorrilla EP, Bartfai T (2008) Phenotypic analysis of GalR2 knockout mice in anxiety- and depression-related behavioral tests. Neuropeptides 42:387–397PubMedCrossRefGoogle Scholar
  18. Picciotto MR (2008) Galanin and addiction. Cell Mol Life Sci 65:1872–1879PubMedCrossRefGoogle Scholar
  19. Shi TJ, Hua XY, Lu X, Malkmus S, Kinney J, Holmberg K, Wirz S, Ceccatelli S, Yaksh T, Bartfai T, Hokfelt T (2006) Sensory neuronal phenotype in galanin receptor 2 knockout mice: focus on dorsal root ganglion neurone development and pain behaviour. Eur J Neurosci 23:627–636PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.The Harold L. Dorris Neurological Research Institute, Molecular and Integrative Neurosciences DepartmentThe Scripps Research InstituteLa JollaUSA

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