Chronically available alcohol escalates drinking in mice and a single injection of the immune activator lipopolysaccharide can mimic this effect and result in a persistent increase in alcohol consumption. We hypothesized that chronic alcohol drinking and lipopolysaccharide injections will produce some similar molecular changes that play a role in regulation of alcohol intake. We investigated the molecular mechanisms of chronic alcohol consumption or lipopolysaccharide insult by gene expression profiling in prefrontal cortex and liver of C57BL/6J mice. We identified similar patterns of transcriptional changes among four groups of animals, three consuming alcohol (vs water) in different consumption tests and one injected with lipopolysaccharide (vs. vehicle). The three tests of alcohol consumption are the continuous chronic two bottle choice (Chronic), two bottle choice available every other day (Chronic Intermittent) and limited access to one bottle of ethanol (Drinking in the Dark). Gene expression changes were more numerous and marked in liver than in prefrontal cortex for the alcohol treatments and similar in the two tissues for lipopolysaccharide. Many of the changes were unique to each treatment, but there was significant overlap in prefrontal cortex for Chronic-Chronic Intermittent and for Chronic Intermittent-lipopolysaccharide and in liver all pairs showed overlap. In silico cell-type analysis indicated that lipopolysaccharide had strongest effects on brain microglia and liver Kupffer cells. Pathway analysis detected a prefrontal cortex-based dopamine-related (PPP1R1B, DRD1, DRD2, FOSB, PDNY) network that was highly over-represented in the Chronic Intermittent group, with several genes from the network being also regulated in the Chronic and lipopolysaccharide (but not Drinking in the Dark) groups. Liver showed a CYP and GST centered metabolic network shared in part by all four treatments. We demonstrate common consequences of chronic alcohol consumption and immune activation in both liver and brain and show distinct genomic consequences of different types of alcohol consumption.
Liu J, Lewohl JM, Harris RA, Iyer VR, Dodd PR, et al. (2006) Patterns of gene expression in the frontal cortex discriminate alcoholic from nonalcoholic individuals. Neuropsychopharmacology 31: 1574–1582.
Zhou Z, Yuan Q, Mash DC, Goldman D (2011) Substance-specific and shared transcription and epigenetic changes in the human hippocampus chronically exposed to cocaine and alcohol. Proc Natl Acad Sci U S A 108: 6626–6631.
McBride WJ, Kimpel MW, Schultz JA, McClintick JN, Edenberg HJ, et al. (2010) Changes in gene expression in regions of the extended amygdala of alcohol-preferring rats after binge-like alcohol drinking. Alcohol 44: 171–183.
Wolstenholme JT, Warner JA, Capparuccini MI, Archer KJ, Shelton KL, et al. (2011) Genomic analysis of individual differences in ethanol drinking: evidence for non-genetic factors in C57BL/6 mice. PLoS One 6: e21100.
Mulligan MK, Ponomarev I, Hitzemann RJ, Belknap JK, Tabakoff B, et al. (2006) Toward understanding the genetics of alcohol drinking through transcriptome meta-analysis. Proc Natl Acad Sci U S A 103: 6368–6373.
Blednov YA, Benavidez JM, Geil C, Perra S, Morikawa H, et al. (2011) Activation of inflammatory signaling by lipopolysaccharide produces a prolonged increase of voluntary alcohol intake in mice. Brain Behav Immun 25 Suppl 1S92–S105.
Deaciuc IV, Doherty DE, Burikhanov R, Lee EY, Stromberg AJ, et al. (2004) Large-scale gene profiling of the liver in a mouse model of chronic, intragastric ethanol infusion. J Hepatol 40: 219–227.
Yin HQ, Je YT, Kim M, Kim JH, Kong G, et al. (2009) Analysis of hepatic gene expression during fatty liver change due to chronic ethanol administration in mice. Toxicol Appl Pharmacol 235: 312–320.
Pfefferbaum A, Sullivan EV, Rosenbloom MJ, Mathalon DH, Lim KO (1998) A controlled study of cortical gray matter and ventricular changes in alcoholic men over a 5-year interval. Arch Gen Psychiatry 55: 905–912.
Leeman RF, Heilig M, Cunningham CL, Stephens DN, Duka T, et al. (2010) Ethanol consumption: how should we measure it? Achieving consilience between human and animal phenotypes. Addict Biol 15: 109–124.
Wahlsten D, Bachmanov A, Finn DA, Crabbe JC (2006) Stability of inbred mouse strain differences in behavior and brain size between laboratories and across decades. Proc Natl Acad Sci U S A 103: 16364–16369.
Szabo G, Mandrekar P, Petrasek J, Catalano D (2011) The unfolding web of innate immune dysregulation in alcoholic liver injury. Alcohol Clin Exp Res 35: 782–786.
Blednov YA, Ponomarev I, Geil C, Bergeson S, Koob GF, et al. (2012) Neuroimmune regulation of alcohol consumption: behavioral validation of genes obtained from genomic studies. Addict Biol 17: 108–120.
Meliska CJ, Bartke A, McGlacken G, Jensen RA (1995) Ethanol, nicotine, amphetamine, and aspartame consumption and preferences in C57BL/6 and DBA/2 mice. Pharmacol Biochem Behav 50: 619–626.
Nocjar C, Middaugh LD, Tavernetti M (1999) Ethanol consumption and place-preference conditioning in the alcohol-preferring C57BL/6 mouse: relationship with motor activity patterns. Alcohol Clin Exp Res 23: 683–692.
Rhodes JS, Best K, Belknap JK, Finn DA, Crabbe JC (2005) Evaluation of a simple model of ethanol drinking to intoxication in C57BL/6J mice. Physiol Behav 84: 53–63.
Ponomarev I, Rau V, Eger EI, Harris RA, Fanselow MS (2010) Amygdala transcriptome and cellular mechanisms underlying stress-enhanced fear learning in a rat model of posttraumatic stress disorder. Neuropsychopharmacology 35: 1402–1411.
Harris RA, Osterndorff-Kahanek E, Ponomarev I, Homanics GE, Blednov YA (2011) Testing the silence of mutations: Transcriptomic and behavioral studies of GABA(A) receptor alpha1 and alpha2 subunit knock-in mice. Neurosci Lett 488: 31–35.
Bolstad BM, Irizarry RA, Astrand M, Speed TP (2003) A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19: 185–193.
Smyth GK (2004) Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 3: Article3.
Cahoy JD, Emery B, Kaushal A, Foo LC, Zamanian JL, et al. (2008) A transcriptome database for astrocytes, neurons, and oligodendrocytes: a new resource for understanding brain development and function. J Neurosci 28: 264–278.
Oldham MC, Konopka G, Iwamoto K, Langfelder P, Kato T, et al. (2008) Functional organization of the transcriptome in human brain. Nat Neurosci 11: 1271–1282.
Takahara Y, Takahashi M, Wagatsuma H, Yokoya F, Zhang QW, et al. (2006) Gene expression profiles of hepatic cell-type specific marker genes in progression of liver fibrosis. World J Gastroenterol 12: 6473–6499.
Huang DW, Sherman BT, Lempicki RA (2009a) Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources. Nature Protoc 4: 44–57.
Huang DW, Sherman BT, Lempicki RA (2009b) Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 37: 1–13.
Becker HC, Diaz-Granados JL, Hale RL (1997) Exacerbation of ethanol withdrawal seizures in mice with a history of multiple withdrawal experience. Pharmacol Biochem Behav 57: 179–183.
Vetreno PV, Crews FT (2013) Innate Immune Signaling and Alcoholism. In: Changhai Cui LG, Antonio Noronha, editor. Neural-Immune Interactions in Brain Function and Alcohol Related Disorders. New York: Springer, U.S. 251–278.
Alfonso-Loeches S, Pascual-Lucas M, Blanco AM, Sanchez-Vera I, Guerri C (2010) Pivotal role of TLR4 receptors in alcohol-induced neuroinflammation and brain damage. J Neurosci 30: 8285–8295.
Blanco AM, Guerri C (2007) Ethanol intake enhances inflammatory mediators in brain: role of glial cells and TLR4/IL-1RI receptors. Front Biosci 12: 2616–2630.
Zou J, Crews F (2010) Induction of innate immune gene expression cascades in brain slice cultures by ethanol: key role of NF-kappaB and proinflammatory cytokines. Alcohol Clin Exp Res 34: 777–789.
Maldve RE, Zhang TA, Ferrani-Kile K, Schreiber SS, Lippmann MJ, et al. (2002) DARPP-32 and regulation of the ethanol sensitivity of NMDA receptors in the nucleus accumbens. Nat Neurosci 5: 641–648.
Nairn AC, Svenningsson P, Nishi A, Fisone G, Girault JA, et al. (2004) The role of DARPP-32 in the actions of drugs of abuse. Neuropharmacology 47 Suppl 114–23.
Femenia T, Manzanares J (2012) Increased ethanol intake in prodynorphin knockout mice is associated to changes in opioid receptor function and dopamine transmission. Addict Biol 17: 322–337.
Bazov I, Kononenko O, Watanabe H, Kuntic V, Sarkisyan D, et al. (2011) The endogenous opioid system in human alcoholics: molecular adaptations in brain areas involved in cognitive control of addiction. Addict Biol 10 (1111/j.1369–1600.2011.00366): x.
Taqi MM, Bazov I, Watanabe H, Sheedy D, Harper C, et al. (2011) Prodynorphin CpG-SNPs associated with alcohol dependence: elevated methylation in the brain of human alcoholics. Addict Biol 16: 499–509.
Clarke TK, Ambrose-Lanci L, Ferraro TN, Berrettini WH, Kampman KM, et al. (2012) Genetic association analyses of PDYN polymorphisms with heroin and cocaine addiction. Genes Brain Behav 11: 415–423.
Kovacs KM, Szakall I, O'Brien D, Wang R, Vinod KY, et al. (2005) Decreased oral self-administration of alcohol in kappa-opioid receptor knock-out mice. Alcohol Clin Exp Res 29: 730–738.
Nishi A, Kuroiwa M, Miller DB, O'Callaghan JP, Bateup HS, et al. (2008) Distinct roles of PDE4 and PDE10A in the regulation of cAMP/PKA signaling in the striatum. J Neurosci 28: 10460–10471.
Contet C, Gardon O, Filliol D, Becker JA, Koob GF, et al. (2011) Identification of genes regulated in the mouse extended amygdala by excessive ethanol drinking associated with dependence. Addict Biol 16: 615–619.
