Mulligan Megan K, Mozhui Khyobeni, Pandey Ashutosh K, Smith Maren L, Gong Suzhen, Ingels Jesse, Miles Michael F, Lopez Marcelo F, Lu Lu, Williams Robert W
Department of Genetics, Genomics and Informatics, University of Tennessee Health Science Center, United States.
Department of Genetics, Genomics and Informatics, University of Tennessee Health Science Center, United States.
Alcohol. 2017 Feb;58:61-72. doi: 10.1016/j.alcohol.2016.09.001. Epub 2016 Oct 15.
Genetic factors that influence the transition from initial drinking to dependence remain enigmatic. Recent studies have leveraged chronic intermittent ethanol (CIE) paradigms to measure changes in brain gene expression in a single strain at 0, 8, 72 h, and even 7 days following CIE. We extend these findings using LCM RNA-seq to profile expression in 11 brain regions in two inbred strains - C57BL/6J (B6) and DBA/2J (D2) - 72 h following multiple cycles of ethanol self-administration and CIE. Linear models identified differential expression based on treatment, region, strain, or interactions with treatment. Nearly 40% of genes showed a robust effect (FDR < 0.01) of region, and hippocampus CA1, cortex, bed nucleus stria terminalis, and nucleus accumbens core had the highest number of differentially expressed genes after treatment. Another 8% of differentially expressed genes demonstrated a robust effect of strain. As expected, based on similar studies in B6, treatment had a much smaller impact on expression; only 72 genes (p < 0.01) are modulated by treatment (independent of region or strain). Strikingly, many more genes (415) show a strain-specific and largely opposite response to treatment and are enriched in processes related to RNA metabolism, transcription factor activity, and mitochondrial function. Over 3 times as many changes in gene expression were detected in D2 compared to B6, and weighted gene co-expression network analysis (WGCNA) module comparison identified more modules enriched for treatment effects in D2. Substantial strain differences exist in the temporal pattern of transcriptional neuroadaptation to CIE, and these may drive individual differences in risk of addiction following excessive alcohol consumption.
影响从初次饮酒到酒精依赖转变的遗传因素仍然是个谜。最近的研究利用慢性间歇性乙醇(CIE)范式来测量在CIE后0、8、72小时甚至7天,单一品系大脑基因表达的变化。我们使用激光捕获显微切割RNA测序(LCM RNA-seq)扩展了这些发现,以分析在多次乙醇自我给药和CIE后72小时,两个近交系——C57BL/6J(B6)和DBA/2J(D2)——11个脑区的基因表达情况。线性模型基于处理、脑区、品系或与处理的相互作用来识别差异表达。近40%的基因显示出脑区的显著影响(错误发现率<0.01),处理后海马CA1区、皮质、终纹床核和伏隔核核心的差异表达基因数量最多。另外8%的差异表达基因显示出品系的显著影响。正如在B6中类似研究预期的那样,处理对基因表达的影响要小得多;只有72个基因(p<0.01)受到处理的调节(与脑区或品系无关)。引人注目的是,更多基因(415个)显示出品系特异性且对处理的反应基本相反,并且在与RNA代谢、转录因子活性和线粒体功能相关的过程中富集。与B6相比,在D2中检测到的基因表达变化多出3倍以上,加权基因共表达网络分析(WGCNA)模块比较确定在D2中有更多模块富集了处理效应。在对CIE的转录神经适应的时间模式上存在显著的品系差异,这些差异可能导致过量饮酒后成瘾风险的个体差异。