Association between anxiety and non-coding genetic variants of the galanin neuropeptide

Autoři: Gergely Keszler aff001;  Zsuzsanna Molnár aff001;  Zsolt Rónai aff001;  Mária Sasvári-Székely aff001;  Anna Székely aff002;  Eszter Kótyuk aff002
Působiště autorů: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary aff001;  MTA‐ELTE Lendület Adaptation Research Group, Institute of Psychology, ELTE Eötvös Loránd University, Budapest, Hungary aff002
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article



Galanin, an inhibitory neuropeptide and cotransmitter has long been known to co-localize with noradrenaline and serotonin in the central nervous system. Several human studies demonstrated altered galanin expression levels in major depressive disorder and anxiety. Pharmacological modulation of galanin signaling and transgenic strategies provide further proof for the involvement of the galanin system in the pathophysiology of mood disorders. Little is known, however, on the dynamic regulation of galanin expression at the transcriptional level. The aim of the present study was to seek genetic association of non-coding single nucleotide variations in the galanin gene with anxiety and depression.


Six single nucleotide polymorphisms (SNP) occurring either in the regulatory 5’ or 3’ flanking regions or within intronic sequences of the galanin gene have been genotyped with a high-throughput TaqMan OpenArray qPCR system in 526 healthy students (40% males). Depression and anxiety scores were obtained by filling in the Hospital Anxiety and Depression Scale (HADS) questionnaire. Data were analyzed by ANCOVA and Bonferroni correction was applied for multiple testing. Linkage disequilibrium (LD) analysis was used to map two haploblocks in the analyzed region.

Results and conclusions

A single-locus and a haplotype genetic association proved to be statistically significant. In single-marker analysis, the T allele of the rs1042577 SNP within the 3’ untranslated region of the galanin gene associated with greater levels of anxiety (HADS scores were 7.05±4.0 vs 6.15±.15; p = 0.000407). Haplotype analysis revealed an association of the rs948854 C_rs4432027_C allele combination with anxiety [F(1,1046) = 4.140, p = 0.042141, η2 = 0.004, power = 0.529]. Neither of these associations turned out to be gender-specific.

These promoter polymorphisms are supposed to participate in epigenetic regulation of galanin expression by creating potentially methylatable CpG dinucleotides. The functional importance of the rs1042577_T allele remains to be elucidated.

Klíčová slova:

Alleles – Anxiety – Depression – DNA methylation – Emotions – Haplotypes – Linkage disequilibrium – Molecular genetics


1. Lang R, Gundlach AL, Holmes FE, Hobson SA, Wynick D, Hökfelt T, et al. Physiology, signaling, and pharmacology of galanin peptides and receptors: three decades of emerging diversity. Pharmacol Rev. 2015;67(1):118–75. doi: 10.1124/pr.112.006536 25428932

2. Hökfelt T, Barde S, Xu ZD, Kuteeva E, Rüegg J, Le Maitre E, et al. Neuropeptide and Small Transmitter Coexistence: Fundamental Studies and Relevance to Mental Illness. Front Neural Circuits. 2018;12:106. doi: 10.3389/fncir.2018.00106 30627087

3. Juhasz G, Hullam G, Eszlari N, Gonda X, Antal P, Anderson IM, et al. Brain galanin system genes interact with life stresses in depression-related phenotypes. Proc Natl Acad Sci U S A. 2014;111(16):E1666–73. doi: 10.1073/pnas.1403649111 24706871

4. Kuteeva E, Hökfelt T, Wardi T, Ogren SO. Galanin, galanin receptor subtypes and depression-like behaviour. Cell Mol Life Sci. 2008;65(12):1854–63. doi: 10.1007/s00018-008-8160-9 18500640

5. de Souza MM, Silote GP, Herbst LS, Funck VR, Joca SRL, Beijamini V. The antidepressant-like effect of galanin in the dorsal raphe nucleus of rats involves GAL2 receptors. Neurosci Lett. 2018;681:26–30. doi: 10.1016/j.neulet.2018.05.029 29787787

6. Lu X, Barr AM, Kinney JW, Sanna P, Conti B, Behrens MM, et al. A role for galanin in antidepressant actions with a focus on the dorsal raphe nucleus. Proc Natl Acad Sci U S A. 2005;102(3):874–9. doi: 10.1073/pnas.0408891102 15647369

7. Bing O, Möller C, Engel JA, Söderpalm B, Heilig M. Anxiolytic-like action of centrally administered galanin. Neurosci Lett. 1993;164(1–2): 17–20. doi: 10.1016/0304-3940(93)90846-d 7512244

8. Möller C, Sommer W, Thorsell A, Heilig M. Anxiogenic-like action of galanin after intra-amygdala administration in the rat. Neuropsychopharmacology. 1999;21(4):507–12. doi: 10.1016/S0893-133X(98)00102-X 10481834

9. Barrera G, Hernandez A, Poulin JF, Laforest S, Drolet G, Morilak DA. Galanin-mediated anxiolytic effect in rat central amygdala is not a result of corelease from noradrenergic terminals. Synapse. 2006;59(1):27–40. doi: 10.1002/syn.20208 16237681

10. Silote GP, Rosal AB, de Souza MM, Beijamini V. Infusion of galanin into the mid-caudal portion of the dorsal raphe nucleus has an anxiolytic effect on rats in the elevated T-maze. Behav Brain Res. 2013;252:312–7. doi: 10.1016/j.bbr.2013.06.023 23791934

11. Funck VR, Fracalossi MP, Vidigal APP, Beijamini V. Dorsal hippocampal galanin modulates anxiety-like behaviours in rats. Brain Res. 2018;1687:74–81. doi: 10.1016/j.brainres.2018.02.036 29499176

12. Morais JS, Souza MM, Campanha TM, Muller CJ, Bittencourt AS, Bortoli VC, et al. Galanin subtype 1 and subtype 2 receptors mediate opposite anxiety-like effects in the rat dorsal raphe nucleus. Behav Brain Res. 2016;314:125–33. doi: 10.1016/j.bbr.2016.08.007 27498247

13. Bailey KR, Pavlova MN, Rohde AD, Hohmann JG, Crawley JN. Galanin receptor subtype 2 (GalR2) null mutant mice display an anxiogenic-like phenotype specific to the elevated plus-maze. Pharmacol Biochem Behav. 2007;86(1):8–20. doi: 10.1016/j.pbb.2006.11.024 17257664

14. Brunner SM, Farzi A, Locker F, Holub BS, Drexel M, Reichmann F, et al. GAL3 receptor KO mice exhibit an anxiety-like phenotype. Proc Natl Acad Sci U S A. 2014;111(19):7138–43. doi: 10.1073/pnas.1318066111 24782539

15. Gonda X, Hullam G, Antal P, Eszlari N, Petschner P, Hökfelt TG, et al. Significance of risk polymorphisms for depression depends on stress exposure. Sci Rep. 2018;8(1):3946. doi: 10.1038/s41598-018-22221-z 29500446

16. Barnabas K, Zhang L, Wang H, Kirouac G, Vrontakis M. Changes in galanin systems in a rat model of post-traumatic stress disorder (PTSD). PLoS One. 2016;11(12):e0167569. doi: 10.1371/journal.pone.0167569 27907151

17. Alexandris A, Liu AK, Chang RC, Pearce RK, Gentleman SM. Differential expression of galanin in the cholinergic basal forebrain of patients with Lewy body disorders. Acta Neuropathol Commun. 2015;3:77. doi: 10.1186/s40478-015-0249-4 26621352

18. Wang YJ, Yang YT, Li H, Liu PZ, Wang CY, Xu ZQ. Plasma galanin is a biomarker for severity of major depressive disorder. Int J Psychiatry Med. 2014;48(2):109–19. doi: 10.2190/PM.48.2.d 25377152

19. Murck H, Held K, Ziegenbein M, Künzel H, Holsboer F, Steiger A. Intravenous administration of the neuropeptide galanin has fast antidepressant efficacy and affects the sleep EEG. Psychoneuroendocrinology. 2004;29(9):1205–11. doi: 10.1016/j.psyneuen.2004.02.006 15219645

20. Barde S, Rüegg J, Prud'homme J, Ekström TJ, Palkovits M, Turecki G, et al. Alterations in the neuropeptide galanin system in major depressive disorder involve levels of transcripts, methylation, and peptide. Proc Natl Acad Sci U S A. 2016;113(52):E8472–E8481. doi: 10.1073/pnas.1617824113 27940914

21. Gresle MM, Butzkueven H, Perreau VM, Jonas A, Xiao J, Thiem S, et al. Galanin is an autocrine myelin and oligodendrocyte trophic signal induced by leukemia inhibitory factor. Glia. 2015;63(6):1005–20. doi: 10.1002/glia.22798 25639936

22. Holmes PV, Yoo HS, Dishman RK. Voluntary exercise and clomipramine treatment elevate prepro-galanin mRNA levels in the locus coeruleus in rats. Neurosci Lett. 2006;408(1):1–4. doi: 10.1016/j.neulet.2006.04.057 16996684

23. Sciolino NR, Dishman RK, Holmes PV. Voluntary exercise offers anxiolytic potential and amplifies galanin gene expression in the locus coeruleus of the rat. Behav Brain Res. 2012;233(1):191–200. doi: 10.1016/j.bbr.2012.05.001 22580167

24. Unschuld PG, Ising M, Erhardt A, Lucae S, Kohli M, Kloiber S, et al. Polymorphisms in the galanin gene are associated with symptom-severity in female patients suffering from panic disorder. J Affect Disord. 2008;105(1–3):177–84. doi: 10.1016/j.jad.2007.05.006 17573119

25. Davidson S, Lear M, Shanley L, Hing B, Baizan-Edge A, Herwig A, et al. Differential activity by polymorphic variants of a remote enhancer that supports galanin expression in the hypothalamus and amygdala: implications for obesity, depression and alcoholism. Neuropsychopharmacology. 2011;36(11):2211–21. doi: 10.1038/npp.2011.93 21716262

26. da Conceição Machado F, de Souza LV, Rangel M, Jara ZP, do Carmo Franco M. The antidepressant-like effect of galanin in the dorsal raphe nucleus of rats involves GAL2 receptors. Neurosci Lett. 2018;681:26–30. doi: 10.1016/j.neulet.2018.05.029 29787787

27. Unschuld PG, Ising M, Roeske D, Erhardt A, Specht M, Kloiber S, et al. Gender-specific association of galanin polymorphisms with HPA-axis dysregulation, symptom severity, and antidepressant treatment response. Neuropsychopharmacology. 2010;35(7):1583–92. doi: 10.1038/npp.2010.30 20237460

28. Wang YJ, Li H, Yang YT, Tie CL, Li F, Xu ZQ, et al. Association of galanin and major depressive disorder in the Chinese Han population. PLoS One. 2013;8(5):e64617. doi: 10.1371/journal.pone.0064617 23741354

29. Kotyuk E, Keszler G, Nemeth N, Ronai Z, Sasvari-Szekely M, Szekely A. Glial cell line-derived neurotrophic factor (GDNF) as a novel candidate gene of anxiety. PLoS One. 2013;8(12):e80613. doi: 10.1371/journal.pone.0080613 24324616

30. Muszbek K, Szekely A, Balogh EM, Molnár M, Rohánszky M, Ruzsa A, et al. Validation of the Hungarian translation of Hospital Anxiety and Depression Scale. Qual Life Res. 2006;15(4):761–6. doi: 10.1007/s11136-005-3627-8 16688508

31. Zigmond AS, Snaith RP. The hospital anxiety and depression scale. Acta Psychiatr Scand. 1983;67(6):361–370. doi: 10.1111/j.1600-0447.1983.tb09716.x 6880820

32. Caci H, Baylé FJ, Mattei V, Dossios C, Robert P, Boyer P. How does the Hospital and Anxiety and Depression Scale measure anxiety and depression in healthy subjects? Psychiatry Res. 2003;118(1):89–99. doi: 10.1016/s0165-1781(03)00044-1 12759165

33. Barrett JC, Fry B, Maller J, Daly MJ. Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics. 2005;21(2):263–5. doi: 10.1093/bioinformatics/bth457 15297300

34. Stephens MJ, Donnelly P. A comparison of bayesian methods for haplotype reconstruction from population genotype data. Am J Hum Genet. 2003;73(5):1162–9. doi: 10.1086/379378 14574645

35. Stephens MJ, Smith N, Donnelly P. Documentation for PHASE, version 2.0.2. 2003

36. Stephens M, Smith NJ, Donnelly P. A new statistical method for haplotype reconstruction from population data. Am J Hum Genet. 2001;68(4):978–989. doi: 10.1086/319501 11254454

37. Miller R. G. Simultaneous statistical inference. 2nd ed, Springer-Verlag New York, 1981.

38. Feingold A. Gender differences in personality: a meta-analysis. Psychological Bulletin 1994;116(3):429–456. doi: 10.1037/0033-2909.116.3.429 7809307

39. Andrews B, Hejdenberg J, Wilding J. Student anxiety and depression: comparison of questionnaire and interview assessments. J Affect Disord. 2006 Oct;95(1–3):29–34. Epub 2006 Jun 9. doi: 10.1016/j.jad.2006.05.003 16764939

40. Kebede MA, Anbessie B, Ayano G. Prevalence and predictors of depression and anxiety among medical students in Addis Ababa, Ethiopia. Int J Ment Health Syst. 2019 May 6;13:30. doi: 10.1186/s13033-019-0287-6 31080499

41. Gan GG, Yuen Ling H. Anxiety, depression and quality of life of medical students in Malaysia. Med J Malaysia. 2019 Feb;74(1):57–61. 30846664

42. Kofler B, Evans HF, Liu ML, Falls V, Iismaa TP, Shine J, et al. Characterization of the 5'-flanking region of the human preprogalanin gene. DNA Cell Biol. 1995;14(4):321–9. doi: 10.1089/dna.1995.14.321 7536007

43. Schäuble N, Reichwald K, Grassl W, Bechstein H, Müller HC, Scherag A, et al. Human galanin (GAL) and galanin 1 receptor (GALR1) variations are not involved in fat intake and early onset obesity. J Nutr. 2005;135(6):1387–92. doi: 10.1093/jn/135.6.1387 15930442

44. Li J, Huang K, Hu G, Babarinde IA, Li Y, Dong X, et al. An alternative CTCF isoform antagonizes canonical CTCF occupancy and changes chromatin architecture to promote apoptosis. Nat Commun. 2019;10(1):1535. doi: 10.1038/s41467-019-08949-w 30948729

Článek vyšel v časopise


2019 Číslo 12
Nejčtenější tento týden