Dual-trait selection for ethanol consumption and withdrawal: genetic and transcriptional network effects.

BACKGROUND Data from C57BL/6J (B6) × DBA/2J (D2) F2 intercrosses (B6xD2 F2 ), standard and recombinant inbred strains, and heterogeneous stock mice indicate that a reciprocal (or inverse) genetic relationship exists between alcohol consumption and withdrawal severity. Furthermore, some genetic studies have detected reciprocal quantitative trait loci (QTLs) for these traits. We used a novel mouse model developed by simultaneous selection for both high alcohol consumption/low withdrawal and low alcohol consumption/high withdrawal and analyzed the gene expression and genome-wide genotypic differences. METHODS Randomly chosen third selected generation (S3 ) mice (N = 24/sex/line), bred from a B6xD2 F2 , were genotyped using the Mouse Universal Genotyping Array, which provided 2,760 informative markers. QTL analysis used a marker-by-marker strategy with the threshold for a significant log of the odds (LOD) set at 10. Gene expression in the ventral striatum was measured using the Illumina Mouse 8.2 array. Differential gene expression and the weighted gene co-expression network analysis (WGCNA) were implemented. RESULTS Significant QTLs for consumption/withdrawal were detected on chromosomes (Chr) 2, 4, 9, and 12. A suggestive QTL mapped to Chr 6. Some of the QTLs overlapped with known QTLs mapped for 1 of the traits individually. One thousand seven hundred and forty-five transcripts were detected as being differentially expressed between the lines; there was some overlap with known withdrawal genes (e.g., Mpdz) located within QTL regions. WGCNA revealed several modules of co-expressed genes showing significant effects in both differential expression and intramodular connectivity; a module richly annotated with kinase-related annotations was most affected. CONCLUSIONS Marked effects of selection on expression and network structure were detected. QTLs overlapping with differentially expressed genes on Chr 2 (distal) and 4 suggest that these are cis-eQTLs (Chr 2: Kif3b, Kcnq2; Chr 4: Mpdz, Snapc3). Other QTLs identified were on Chr 2 (proximal), 9, and 12. Network results point to involvement of kinase-related mechanisms and outline the need for further efforts such as interrogation of noncoding RNAs.

[1]  S. Grant,et al.  Novel MPDZ/MUPP1 transgenic and knockdown models confirm Mpdz's role in ethanol withdrawal and support its role in voluntary ethanol consumption , 2015, Addiction biology.

[2]  U. Heberlein,et al.  Happyhour, a Ste20 Family Kinase, Implicates EGFR Signaling in Ethanol-Induced Behaviors , 2014, Cell.

[3]  J. Crabbe,et al.  Selection for drinking in the dark alters brain gene coexpression networks. , 2013, Alcoholism, clinical and experimental research.

[4]  A. Barkley-Levenson,et al.  Ethanol drinking in withdrawal seizure-prone and -resistant selected mouse lines. , 2013, Alcohol.

[5]  S. Kang,et al.  PI3-Kinase γ Promotes Rap1a-Mediated Activation of Myeloid Cell Integrin α4β1, Leading to Tumor Inflammation and Growth , 2013, PloS one.

[6]  Shannon McWeeney,et al.  Differential Network Analysis Reveals Genetic Effects on Catalepsy Modules , 2013, PloS one.

[7]  D. Bottomly,et al.  Genes, behavior and next‐generation RNA sequencing , 2013, Genes, brain, and behavior.

[8]  P. Janak,et al.  BDNF‐mediated regulation of ethanol consumption requires the activation of the MAP kinase pathway and protein synthesis , 2013, The European journal of neuroscience.

[9]  Daniel Bottomly,et al.  Utilizing RNA-Seq data for de novo coexpression network inference , 2012, Bioinform..

[10]  S. McWeeney,et al.  Gene networks and haloperidol‐induced catalepsy , 2012, Genes, brain, and behavior.

[11]  Megan K. Mulligan,et al.  Molecular profiles of drinking alcohol to intoxication in C57BL/6J mice. , 2011, Alcoholism, clinical and experimental research.

[12]  Rui Luo,et al.  Is My Network Module Preserved and Reproducible? , 2011, PLoS Comput. Biol..

[13]  Shannon McWeeney,et al.  Genetic diversity and striatal gene networks: focus on the heterogeneous stock-collaborative cross (HS-CC) mouse , 2010, BMC Genomics.

[14]  Nora D Volkow,et al.  Neurocircuitry of Addiction , 2010, Neuropsychopharmacology.

[15]  W. Mcbride,et al.  Gene expression changes in the nucleus accumbens of alcohol-preferring rats following chronic ethanol consumption , 2009, Pharmacology Biochemistry and Behavior.

[16]  U. Heberlein,et al.  Happyhour, a Ste20 Family Kinase, Implicates EGFR Signaling in Ethanol-Induced Behaviors , 2009, Cell.

[17]  S. McWeeney,et al.  Detection of reciprocal quantitative trait loci for acute ethanol withdrawal and ethanol consumption in heterogeneous stock mice , 2009, Psychopharmacology.

[18]  C. Hodge,et al.  Increased operant responding for ethanol in male C57BL/6J mice: specific regulation by the ERK1/2, but not JNK, MAP kinase pathway , 2009, Psychopharmacology.

[19]  Steve Horvath,et al.  WGCNA: an R package for weighted correlation network analysis , 2008, BMC Bioinformatics.

[20]  L. Kaczmarek,et al.  Alcohol Relapse Induced by Discrete Cues Activates Components of AP-1 Transcription Factor and ERK Pathway in the Rat Basolateral and Central Amygdala , 2008, Neuropsychopharmacology.

[21]  A. Hansson,et al.  Neuroplasticity in brain reward circuitry following a history of ethanol dependence , 2008, The European journal of neuroscience.

[22]  Y. Xing,et al.  A Transcriptome Database for Astrocytes, Neurons, and Oligodendrocytes: A New Resource for Understanding Brain Development and Function , 2008, The Journal of Neuroscience.

[23]  S. Horvath,et al.  Weighted gene coexpression network analysis strategies applied to mouse weight , 2007, Mammalian Genome.

[24]  Ming D. Li,et al.  Regulation of platelet-derived growth factor signaling pathway by ethanol, nicotine, or both in mouse cortical neurons. , 2007, Alcoholism, clinical and experimental research.

[25]  P. Hyytiä,et al.  REVIEW: The alcohol‐preferring AA and alcohol‐avoiding ANA rats: neurobiology of the regulation of alcohol drinking , 2006, Addiction biology.

[26]  Megan K. Mulligan,et al.  Toward understanding the genetics of alcohol drinking through transcriptome meta-analysis. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[27]  S. Horvath,et al.  A General Framework for Weighted Gene Co-Expression Network Analysis , 2005, Statistical applications in genetics and molecular biology.

[28]  J. Crabbe,et al.  Alcohol withdrawal severity in inbred mouse (Mus musculus) strains. , 2005, Behavioral neuroscience.

[29]  N. Walter,et al.  Mpdz is a quantitative trait gene for drug withdrawal seizures , 2004, Nature Neuroscience.

[30]  J. A. Chester,et al.  Further evidence of an inverse genetic relationship between innate differences in alcohol preference and alcohol withdrawal magnitude in multiple selectively bred rat lines. , 2003, Alcoholism, clinical and experimental research.

[31]  J. Crabbe,et al.  Mapping murine loci for physical dependence on ethanol , 2002, Psychopharmacology.

[32]  J. Belknap,et al.  The replicability of QTLs for murine alcohol preference drinking behavior across eight independent studies , 2001, Mammalian Genome.

[33]  G. Mcclearn,et al.  High genetic susceptibility to ethanol withdrawal predicts low ethanol consumption , 1998, Mammalian Genome.

[34]  J. Belknap,et al.  Genes on mouse Chromosomes 2 and 9 determine variation in ethanol consumption , 1998, Mammalian Genome.

[35]  J. Crabbe,et al.  Drug withdrawal convulsions and susceptibility to convulsants after short-term selective breeding for acute ethanol withdrawal , 1998, Behavioural Brain Research.

[36]  J. Crabbe,et al.  Provisional mapping of quantitative trait loci for chronic ethanol withdrawal severity in BXD recombinant inbred mice. , 1998, The Journal of pharmacology and experimental therapeutics.

[37]  J. Crabbe,et al.  Quantitative Trait Loci Involved in Genetic Predisposition to Acute Alcohol Withdrawal in Mice , 1997, The Journal of Neuroscience.

[38]  J. Crabbe,et al.  Localization of genes affecting alcohol drinking in mice. , 1994, Alcoholism, clinical and experimental research.

[39]  J. Crabbe,et al.  Voluntary consumption of ethanol in 15 inbred mouse strains , 1993, Psychopharmacology.

[40]  J. Crabbe,et al.  Voluntary consumption of ethanol in WSP, WSC and WSR selectively bred mouse lines , 1988, Pharmacology, Biochemistry and Behavior.

[41]  V. Dole,et al.  Toward an analogue of alcoholism in mice: scale factors in the model. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[42]  D. B. Goldstein.  Relationship of alcohol dose to intensity of withdrawal signs in mice. , 1972, The Journal of pharmacology and experimental therapeutics.

[43]  D. B. Goldstein,et al.  Alcohol Dependence Produced in Mice by Inhalation of Ethanol: Grading the Withdrawal Reaction , 1971, Science.

[44]  J. Ballenger.  GDNF is a fast-acting potent inhibitor of alcohol consumption and relapse , 2009 .

[45]  Lon R. Cardon,et al.  Quantitative Trait Loci , 1995 .

[46]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[47]  J. Crabbe,et al.  Use of recombinant inbred strains for studying genetic determinants of responses to alcohol. , 1994, Alcohol and alcoholism (Oxford, Oxfordshire). Supplement.