Alcohol use disorder is one of the most disabling diseases worldwide. Glial-cell derived neurotrophic factor (Gdnf) shows promising results concerning the inhibition of alcohol consumption in rodent models. We investigated the epigenetic regulation of Gdnf following ethanol consumption and withdrawal in a rat model. 32 Wistar rats underwent 7 weeks of intermittent access to alcohol in a 2-bottle choice (IA2BC). Whole blood, Nucleus Accumbens (NAc) and Ventral Tegmental Area (VTA) were collected immediately after the last 24 h of an alcohol-drinking session (alcohol group, AG) or 24 h after withdrawal (withdrawal group, WG). MRNA levels were measured using real-time quantitative PCR. Bisulfite-conversion of DNA and capillary sequencing was used to determine methylation levels of the core promoter (CP) and the negative regulatory element (NRE). The CP of the AG in the NAc was significantly less methylated compared to controls (p < 0.05). In the NAc, mRNA expression was significantly higher in the WG (p < 0.05). In the WG, mRNA expression levels in the VTA were significantly lower (p < 0.05) and showed significantly less methylation in the NRE in the VTA (p < 0.001) and the NAc (p < 0.01) compared to controls. Changes in the cerebral mRNA expression correspond to alterations in DNA methylation of the Gdnf promoter in a rodent model. Our results hold clinical relevance since differences in Gdnf mRNA expression and DNA methylation could be a target for pharmacological interventions.

中文翻译：

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饮酒会改变大鼠Gdnf启动子的甲基化和表达。

饮酒障碍是全球最致残的疾病之一。胶质细胞衍生的神经营养因子（Gdnf）在啮齿类动物模型中显示出关于抑制饮酒的令人鼓舞的结果。我们调查了大鼠模型中乙醇消耗和戒断后Gdnf的表观遗传调控。在2瓶选择（IA2BC）中，对32只Wistar大鼠进行了7周间歇性饮酒。饮酒后最后24小时（酒精组，AG）或戒断后24小时（戒断组，WG）收集全血，伏隔核（NAc）和腹侧被盖区（VTA）。使用实时定量PCR测量MRNA水平。DNA的亚硫酸氢盐转化和毛细管测序用于确定核心启动子（CP）和负调控元件（NRE）的甲基化水平。与对照组相比，NAc中AG的CP甲基化程度显着降低（p <0.05）。在NAc中，WG中的mRNA表达明显更高（p <0.05）。与对照组相比，在WG中，VTA中的mRNA表达水平显着降低（p <0.05），并且显示VTA中的NRE（p <0.001）和NAc（p <0.01）中的甲基化水平显着降低。啮齿动物模型中脑mRNA表达的变化与Gdnf启动子DNA甲基化的变化相对应。我们的结果具有临床意义，因为Gdnf mRNA表达和DNA甲基化的差异可能是药物干预的目标。与对照组相比，VTA中的mRNA表达水平显着降低（p <0.05），并且NTA中的NRE（p <0.001）和NAc（p <0.01）中的甲基化水平显着降低。啮齿动物模型中脑mRNA表达的变化与Gdnf启动子DNA甲基化的变化相对应。我们的结果具有临床意义，因为Gdnf mRNA表达和DNA甲基化的差异可能是药物干预的目标。与对照组相比，VTA中的mRNA表达水平显着降低（p <0.05），并且NTA中的NRE（p <0.001）和NAc（p <0.01）中的甲基化水平显着降低。啮齿动物模型中脑mRNA表达的变化与Gdnf启动子DNA甲基化的变化相对应。我们的结果具有临床意义，因为Gdnf mRNA表达和DNA甲基化的差异可能是药物干预的目标。