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Genetic and genomic signatures in ethanol withdrawal seizure-prone and seizure-resistant mice implicate genes involved in epilepsy and neuronal excitability

Molecular Psychiatry volume 27, pages 4611–4623 (2022)Cite this article

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Abstract

Alcohol withdrawal is a clinically important consequence and potential driver of Alcohol Use Disorder. However, susceptibility to withdrawal symptoms, ranging from craving and anxiety to seizures and delirium, varies greatly. Selectively bred Withdrawal Seizure-Prone (WSP) and Seizure-Resistant (WSR) mice are an animal model of differential susceptibility to withdrawal and phenotypes with which withdrawal severity correlates. To identify innate drivers of alcohol withdrawal severity, we performed a multi-omic study of the WSP and WSR lines and F2 mice derived from them, using genomic, genetic, and transcriptomic analyses. Genes implicated in seizures and epilepsy were over-represented among those that segregated between WSP and WSR mice and that displayed differential expression in F2 mice high and low in withdrawal. Quantitative trait locus (QTL) analysis of ethanol withdrawal convulsions identified several genome-wide significant loci and pointed to genes that modulate potassium channel function and neural excitability. Perturbations of expression of genes involved in synaptic transmission, including GABAergic and glutamatergic genes, were prominent in prefrontal cortex transcriptome. Expression QTL (eQTL) analysis fine mapped genes within the peak ethanol withdrawal QTL regions. Genetic association analysis in human subjects provided converging evidence for the involvement of those genes in severity of alcohol withdrawal and dependence. Our results reveal a polygenic network and neural signaling pathways contributing to ethanol withdrawal seizures and related phenotypes that overlap with genes modulating epilepsy and neuronal excitability.

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Fig. 1: Genetic differences, QTL mapping, and transcriptome analysis of selectively bred WSP/WSR mice.
Fig. 2: Over-representation of seizure-related genes among genes either co-segregating or showing differential expression between seizure-prone and seizure-resistant mice.
Fig. 3: Perturbed expression of genes involved in GABAergic and glutamatergic transmission is implicated in ethanol withdrawal convulsion.
Fig. 4: Genome-wide identification of cis-regulated gene expression and co-expression networks.
Fig. 5: Relationships of cis-regulated gene expression, ethanol withdrawal phenotype-based differential expression, and withdrawal QTL signals at three loci.

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Data availability

Sequences of the WSP1, WSR1, WSP2, and WSR2 exomes and WSP/WSR F2 transcriptomes have been deposited in NCBI as a BioProject (accession no. PRJNA875906).

References

  1. Hall W, Zador D. The alcohol withdrawal syndrome. Lancet. 1997;349:1897–1900.

    CAS  PubMed  Google Scholar 

  2. Gilpin NW, Koob GF. Neurobiology of alcohol dependence: focus on motivational mechanisms. Alcohol Res Health. 2008;31:185–95.

    PubMed  PubMed Central  Google Scholar 

  3. Goldman D, Oroszi G, Ducci F. The genetics of addictions: uncovering the genes. Nat Rev Genet. 2005;6:521–32.

    CAS  PubMed  Google Scholar 

  4. Edenberg HJ, Koller DL, Xuei X, Wetherill L, McClintick JN, Almasy L, et al. Genome-wide association study of alcohol dependence implicates a region on chromosome 11. Alcohol Clin Exp Res. 2010;34:840–52.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Walters RK, Polimanti R, Johnson EC, McClintick JN, Adams MJ, Adkins AE, et al. Transancestral GWAS of alcohol dependence reveals common genetic underpinnings with psychiatric disorders. Nat Neurosci. 2018;21:1656–69.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Zhou Z, Karlsson C, Liang T, Xiong W, Kimura M, Tapocik JD, et al. Loss of metabotropic glutamate receptor 2 escalates alcohol consumption. Proc Natl Acad Sci USA. 2013;110:16963–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Zhou Z, Yuan Q, Mash DC, Goldman D. Substance-specific and shared transcription and epigenetic changes in the human hippocampus chronically exposed to cocaine and alcohol. Proc Natl Acad Sci USA. 2011;108:6626–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Mhatre MC, Ticku MK. Chronic ethanol administration alters gamma-aminobutyric acid A receptor gene expression. Mol Pharm. 1992;42:415–22.

    CAS  Google Scholar 

  9. Herman MA, Roberto M. Cell-type-specific tonic GABA signaling in the rat central amygdala is selectively altered by acute and chronic ethanol. Addict Biol. 2016;21:72–86.

    CAS  PubMed  Google Scholar 

  10. Christian DT, Alexander NJ, Diaz MR, McCool BA. Thalamic glutamatergic afferents into the rat basolateral amygdala exhibit increased presynaptic glutamate function following withdrawal from chronic intermittent ethanol. Neuropharmacology. 2013;65:134–42.

    CAS  PubMed  Google Scholar 

  11. Griffin WC 3rd, Haun HL, Hazelbaker CL, Ramachandra VS, Becker HC. Increased extracellular glutamate in the nucleus accumbens promotes excessive ethanol drinking in ethanol-dependent mice. Neuropsychopharmacology. 2014;39:707–17.

    CAS  PubMed  Google Scholar 

  12. Abrahao KP, Salinas AG, Lovinger DM. Alcohol and the brain: neuronal molecular targets, synapses, and circuits. Neuron. 2017;96:1223–38.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Tabarki B, AlMajhad N, AlHashem A, Shaheen R, Alkuraya FS. Homozygous KCNMA1 mutation as a cause of cerebellar atrophy, developmental delay and seizures. Hum Genet. 2016;135:1295–8.

    CAS  PubMed  Google Scholar 

  14. Krapivinsky G, Medina I, Krapivinsky L, Gapon S, Clapham DE. SynGAP-MUPP1-CaMKII synaptic complexes regulate p38 MAP kinase activity and NMDA receptor-dependent synaptic AMPA receptor potentiation. Neuron. 2004;43:563–74.

    CAS  PubMed  Google Scholar 

  15. Fehr C, Shirley RL, Belknap JK, Crabbe JC, Buck KJ. Congenic mapping of alcohol and pentobarbital withdrawal liability loci to a <1 centimorgan interval of murine chromosome 4: identification of Mpdz as a candidate gene. J Neurosci. 2002;22:3730–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Bubier JA, Jay JJ, Baker CL, Bergeson SE, Ohno H, Metten P, et al. Identification of a QTL in Mus musculus for alcohol preference, withdrawal, and Ap3m2 expression using integrative functional genomics and precision genetics. Genetics. 2014;197:1377–93.

    PubMed  PubMed Central  Google Scholar 

  17. Crabbe JC, Kosobud A, Young ER, Tam BR, McSwigan JD. Bidirectional selection for susceptibility to ethanol withdrawal seizures in Mus musculus. Behav Genet. 1985;15:521–36.

    CAS  PubMed  Google Scholar 

  18. Metten P, Schlumbohm JP, Huang LC, Greenberg GD, Hack WR, Spence SE, et al. An alcohol withdrawal test battery measuring multiple behavioral symptoms in mice. Alcohol. 2018;68:19–35.

    PubMed  Google Scholar 

  19. McClearn GE, Wilson JR, Meredith JE. The use of isogenic and heterogenic mouse stocks in behavioral research. In: G Lindzey DT (ed). Contributions to behavior-genetic analysis: the mouse as a prototype. Appleton-Century-Crofts: New York, 1970, pp 3–22.

  20. Crabbe JC, Spence SE, Huang LC, Cameron AJ, Schlumbohm JP, Barkley-Levenson AM, et al. Ethanol drinking in Withdrawal Seizure-Prone and -Resistant selected mouse lines. Alcohol. 2013;47:381–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Hartmann MC, Holbrook SE, Haney MM, Crabbe JC, Rosenwasser AM. Affective behavior in withdrawal seizure-prone and withdrawal seizure-resistant mice during long-term alcohol abstinence. Alcohol Clin Exp Res. 2019;43:1478–85.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Philibin SD, Cameron AJ, Schlumbohm JP, Metten P, Crabbe JC. Ethanol withdrawal-induced motor impairment in mice. Psychopharmacology. 2012;220:367–78.

    CAS  PubMed  Google Scholar 

  23. Bergeson SE, Kyle Warren R, Crabbe JC, Metten P, Gene Erwin V, Belknap JK. Chromosomal loci influencing chronic alcohol withdrawal severity. Mamm Genome. 2003;14:454–63.

    PubMed  Google Scholar 

  24. Mark GP, Finn DA. The relationship between hippocampal acetylcholine release and cholinergic convulsant sensitivity in withdrawal seizure-prone and withdrawal seizure-resistant selected mouse lines. Alcohol Clin Exp Res. 2002;26:1141–52.

    CAS  PubMed  Google Scholar 

  25. Plotkin SR, Banks WA, Cohn CS, Kastin AJ. Withdrawal from alcohol in withdrawal seizure-prone and -resistant mice: evidence for enkephalin resistance. Pharm Biochem Behav. 2001;68:379–87.

    CAS  Google Scholar 

  26. Snelling C, Tanchuck-Nipper MA, Ford MM, Jensen JP, Cozzoli DK, Ramaker MJ, et al. Quantification of ten neuroactive steroids in plasma in Withdrawal Seizure-Prone and -Resistant mice during chronic ethanol withdrawal. Psychopharmacology. 2014;231:3401–14.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Keir WJ, Morrow AL. Differential expression of GABAA receptor subunit mRNAs in ethanol-naive withdrawal seizure resistant (WSR) vs. withdrawal seizure prone (WSP) mouse brain. Brain Res Mol Brain Res. 1994;25:200–8.

    CAS  PubMed  Google Scholar 

  28. Mason JN, Eshleman AJ, Belknap JK, Crabbe JC, Loftis JM, Macey TA, et al. NMDA receptor subunit mRNA and protein expression in ethanol-withdrawal seizure-prone and -resistant mice. Alcohol Clin Exp Res. 2001;25:651–60.

    CAS  PubMed  Google Scholar 

  29. Langfelder P, Horvath S. WGCNA: an R package for weighted correlation network analysis. BMC Bioinform. 2008;9:559.

    Google Scholar 

  30. Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, et al. STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 2019;47:D607–d613.

    CAS  PubMed  Google Scholar 

  31. Ridgway LD, Kim EY, Dryer SE. MAGI-1 interacts with Slo1 channel proteins and suppresses Slo1 expression on the cell surface. Am J Physiol Cell Physiol. 2009;297:C55–65.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Tanemoto M, Abe T, Uchida S, Kawahara K. Mislocalization of K+ channels causes the renal salt wasting in EAST/SeSAME syndrome. FEBS Lett. 2014;588:899–905.

    CAS  PubMed  Google Scholar 

  33. Shirley RL, Walter NA, Reilly MT, Fehr C, Buck KJ. Mpdz is a quantitative trait gene for drug withdrawal seizures. Nat Neurosci. 2004;7:699–700.

    CAS  PubMed  Google Scholar 

  34. Hu JH, Park JM, Park S, Xiao B, Dehoff MH, Kim S, et al. Homeostatic scaling requires group I mGluR activation mediated by Homer1a. Neuron. 2010;68:1128–42.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Kammermeier PJ. Endogenous homer proteins regulate metabotropic glutamate receptor signaling in neurons. J Neurosci. 2008;28:8560–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Medina-Kauwe LK, Tobin AJ, De Meirleir L, Jaeken J, Jakobs C, Nyhan WL, et al. 4-Aminobutyrate aminotransferase (GABA-transaminase) deficiency. J Inherit Metab Dis. 1999;22:414–27.

    CAS  PubMed  Google Scholar 

  37. Youle RJ, van der Bliek AM. Mitochondrial fission, fusion, and stress. Science. 2012;337:1062–5.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Tonduti D, Dorboz I, Imbard A, Slama A, Boutron A, Pichard S, et al. New spastic paraplegia phenotype associated to mutation of NFU1. Orphanet J Rare Dis. 2015;10:13.

    PubMed  PubMed Central  Google Scholar 

  39. Zhang R, Engler A, Taylor V. Notch: an interactive player in neurogenesis and disease. Cell Tissue Res. 2018;371:73–89.

    CAS  PubMed  Google Scholar 

  40. Noebels JL. Sodium channel gene expression and epilepsy. Novartis Found Symp. 2002;241:109–20. discussion 120-103, 226-132

    CAS  PubMed  Google Scholar 

  41. Reynolds C, King MD, Gorman KM. The phenotypic spectrum of SCN2A-related epilepsy. Eur J Paediatr Neurol. 2020;24:117–22.

    PubMed  Google Scholar 

  42. Erol A, Karpyak VM. Sex and gender-related differences in alcohol use and its consequences: contemporary knowledge and future research considerations. Drug Alcohol Depend. 2015;156:1–13.

    PubMed  Google Scholar 

  43. Sharrett-Field L, Butler TR, Reynolds AR, Berry JN, Prendergast MA. Sex differences in neuroadaptation to alcohol and withdrawal neurotoxicity. Pflug Arch. 2013;465:643–54.

    CAS  Google Scholar 

  44. Sullivan JT, Sykora K, Schneiderman J, Naranjo CA, Sellers EM. Assessment of alcohol withdrawal: the revised clinical institute withdrawal assessment for alcohol scale (CIWA-Ar). Br J Addict. 1989;84:1353–7.

    CAS  PubMed  Google Scholar 

  45. Zhou Z, Roy A, Lipsky R, Kuchipudi K, Zhu G, Taubman J, et al. Haplotype-based linkage of tryptophan hydroxylase 2 to suicide attempt, major depression, and cerebrospinal fluid 5-hydroxyindoleacetic acid in 4 populations. Arch Gen Psychiatry. 2005;62:1109–18.

    CAS  PubMed  Google Scholar 

  46. Khalili AA, Loudet O. eqtl: tools for analyzing eQTL experiments: a complementary to Karl Broman’s ‘qtl’ package for genome-wide analysis. 2012.

  47. Broman KW, Wu H, Sen S, Churchill GA. R/qtl: QTL mapping in experimental crosses. Bioinformatics. 2003;19:889–90.

    CAS  PubMed  Google Scholar 

  48. Clayton D. snpStats: SnpMatrix and XSnpMatrix classes and methods. R package version 1.16.0. 2014.

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Acknowledgements

The authors wish to thank many former technicians, interns, post-doctoral fellows, and graduate students of the Crabbe lab whose efforts in breeding, testing, and scoring withdrawal behaviors were invaluable. This work was supported by the Intramural Research Program of the NIAAA; NIAAA Grant AA10760 (to JCC); and US Department of Veteran’s Affairs Grant BX000313 (to JCC).

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  1. These authors contributed equally: Zhifeng Zhou, Pamela Metten.

Authors and Affiliations

  1. Laboratory of Neurogenetics, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD, USA

    Zhifeng Zhou, Qiaoping Yuan, Hui Sun, Colin A. Hodgkinson, Pei-Hong Shen, Cheryl Marietta & David Goldman

  2. VA Portland Health Care System and Department of Behavioral Neuroscience, Oregon Health & Science University, Portland, OR, USA

    Pamela Metten & John C. Crabbe

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Contributions

ZZ, PM, JCC, and DG designed the research project. ZZ, PM, HS, CAH, and CM carried out or contributed to the experiments. ZZ, QY, and P-HS analyzed the data. ZZ, PM, JCC, and DG wrote the paper.

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Correspondence to David Goldman.

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Zhou, Z., Metten, P., Yuan, Q. et al. Genetic and genomic signatures in ethanol withdrawal seizure-prone and seizure-resistant mice implicate genes involved in epilepsy and neuronal excitability. Mol Psychiatry 27, 4611–4623 (2022). https://doi.org/10.1038/s41380-022-01799-x

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