Voluntary alcohol consumption represents an endophenotype that can be modeled in rodents and that may reflect susceptibility to the development of alcohol dependence. Selective breeding of mice and rats for differences in this phenotype indicates a genetic influence on voluntary alcohol consumption. We have used mice selectively bred for high and low alcohol preference (HAP and LAP mice, respectively), as measured in a two-bottle choice paradigm, to identify candidate genes that contribute to alcohol preference drinking through their differential expression levels in brain. We now propose to confirm the differential expression of these genes, and localize the differential expression within brain regions, by quantitative reverse transcriptase real-time PCR (qRT-PCR) using a """"""""voxelation"""""""" procedure, and by quantitative in situ hybridization, in brains of HAP and LAP mice. The expression of selected genes, prioritized based on function and proposed role in alcohol drinking behavior, will then be reduced by treatment of the mice with RNAi reagents. We will design siRNAs and shRNAs in collaboration with the INIA RNAi Core and Dharmacon. The efficacy and specificity of these reagents will be assayed in vitro. siRNAs will be delivered using osmotic minipumps or by site-specific infection of lentiviral vectors containing shRNAs. We will determine the time course, duration and extent of target gene down-regulation by qRT-PCR and specificity of effects by qRT-PCR and microarray analysis in collaboration with the INIA Colorado Gene Array Core. Once specific gene knockdown has been confirmed, mice will be tested for changes in alcohol preference drinking in the two-bottle choice paradigm. HAP and LAP mice will also be treated chronically with ethanol in the """"""""withdrawal-induced drinking"""""""" procedures of the INIA Mouse Animal Models Core. Changes in brain gene expression that correlate with increases in voluntary alcohol consumption, following the chronic alcohol exposure, will be determined. These experiments will systematically investigate the role of identified candidate genes, alone and in combinations reflecting signal transduction pathways, in the modulation of voluntary alcohol consumption. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AA016649-03
Application #
7483227
Study Section
Special Emphasis Panel (ZAA1-DD (70))
Program Officer
Reilly, Matthew
Project Start
2006-09-30
Project End
2011-08-31
Budget Start
2008-09-01
Budget End
2009-08-31
Support Year
3
Fiscal Year
2008
Total Cost
$261,650
Indirect Cost
Name
University of Colorado Denver
Department
Pharmacology
Type
Schools of Medicine
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
Vanderlinden, Lauren A; Saba, Laura M; Bennett, Beth et al. (2015) Influence of sex on genetic regulation of ""drinking in the dark"" alcohol consumption. Mamm Genome 26:43-56
Saba, Laura M; Flink, Stephen C; Vanderlinden, Lauren A et al. (2015) The sequenced rat brain transcriptome--its use in identifying networks predisposing alcohol consumption. FEBS J 282:3556-78
Vanderlinden, Lauren A; Saba, Laura M; Printz, Morton P et al. (2014) Is the alcohol deprivation effect genetically mediated? Studies with HXB/BXH recombinant inbred rat strains. Alcohol Clin Exp Res 38:2148-57
Hoffman, P L; Saba, L M; Flink, S et al. (2014) Genetics of gene expression characterizes response to selective breeding for alcohol preference. Genes Brain Behav 13:743-57
Vanderlinden, Lauren A; Saba, Laura M; Kechris, Katerina et al. (2013) Whole brain and brain regional coexpression network interactions associated with predisposition to alcohol consumption. PLoS One 8:e68878
Tabakoff, Boris; Hoffman, Paula L (2013) The neurobiology of alcohol consumption and alcoholism: an integrative history. Pharmacol Biochem Behav 113:20-37
Hoffman, Paula L; Bennett, Beth; Saba, Laura M et al. (2011) Using the Phenogen website for 'in silico' analysis of morphine-induced analgesia: identifying candidate genes. Addict Biol 16:393-404
Desrivières, Sylvane; Pronko, Sergey P; Lourdusamy, Anbarasu et al. (2011) Sex-specific role for adenylyl cyclase type 7 in alcohol dependence. Biol Psychiatry 69:1100-8
Bennett, Beth; Saba, Laura M; Hornbaker, Cheryl K et al. (2011) Genetical genomic analysis of complex phenotypes using the PhenoGen website. Behav Genet 41:625-8
Saba, Laura M; Bennett, Beth; Hoffman, Paula L et al. (2011) A systems genetic analysis of alcohol drinking by mice, rats and men: influence of brain GABAergic transmission. Neuropharmacology 60:1269-80

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