AIMS Alcohol abuse has attracted public attention and chronic alcohol exposure can result in irreversible structural changes in the brain. The molecular mechanisms underlying alcohol neurotoxicity are complex, mandating comprehensive mining of spatial protein expression profile. METHODS In this study, mice models of chronic alcohol intoxication were established after 95% alcohol vapor administration for 30 consecutive days. On Day 30, striatum (the dorsal and ventral striatum) and hippocampus, the two major brain regions responsible for learning and memorizing while being sensitive to alcohol toxicity, were collected. After that, isobaric tags for relative and absolute quantitation -based quantitative proteomic analysis were carried out for further exploration of the novel mechanisms underlying alcohol neurotoxicity. RESULTS Proteomic results showed that in the striatum, 29 proteins were significantly up-regulated and 17 proteins were significantly down-regulated. In the hippocampus, 72 proteins were significantly up-regulated, while 2 proteins were significantly down-regulated. Analysis of the overlay proteins revealed that a total of 102 proteins were consistently altered (P < 0.05) in both hippocampus and striatum regions, including multiple keratins such as Krt6a, Krt17 and Krt5. Ingenuity pathway analysis revealed that previously reported diseases/biofunctions such as dermatological diseases and developmental disorders were enriched in those proteins. Interestingly, the glucocorticoid receptor (GR) signaling was among the top enriched pathways in both brain regions, while multiple keratins from the GR signaling such as Krt1 and Krt17 exhibited significantly opposite expression patterns in the two brain nuclei. Moreover, there are several other involved pathways significantly differed between the hippocampus and striatum. CONCLUSIONS Our data revealed brain regional differences upon alcohol consumption and indicated the critical involvement of keratins from GR signaling in alcohol neurotoxicity. The differences in proteomic results between the striatum and hippocampus suggested a necessity of taking into consideration brain regional differences and intertwined signaling pathways rather than merely focusing on single nuclei or molecule during the study of drug-induced neurotoxicity in the future.

中文翻译：

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大脑区域的蛋白质组学分析揭示了大脑区域差异以及慢性酒精神经毒性中多种角蛋白的参与。

AIMS酗酒引起了公众的关注，长期酗酒会导致大脑中不可逆转的结构变化。酒精神经毒性的潜在分子机制很复杂，要求全面挖掘空间蛋白质表达谱。方法在这项研究中，连续95天给予95％酒精蒸气后，建立了慢性酒精中毒的小鼠模型。在第30天，收集纹状体（背侧和腹侧纹状体）和海马，这两个主要的大脑区域负责学习和记忆，同时对酒精中毒敏感。此后，进行了基于相对定量和绝对定量的蛋白质组学分析的同量异位标记，以进一步探索酒精神经毒性的新机制。结果蛋白质组学结果显示，纹状体中29种蛋白显着上调，17种蛋白显着下调。在海马中，有72种蛋白被显着上调，而有2种蛋白被显着下调。对覆盖蛋白的分析表明，在海马和纹状体区域中共有102种蛋白质被一致地改变（P <0.05），包括多种角蛋白，如Krt6a，Krt17和Krt5。创造力途径分析表明，以前报道的疾病/生物功能（如皮肤病和发育障碍）富含这些蛋白质。有趣的是，糖皮质激素受体（GR）信号是两个大脑区域中最富集的途径之一，而来自GR信号的多个角蛋白（例如Krt1和Krt17）在两个脑核中表现出明显相反的表达模式。此外，海马和纹状体之间还有其他明显不同的参与途径。结论我们的数据揭示了饮酒后大脑区域的差异，并表明了GR信号转导的角蛋白在酒精神经毒性中的关键作用。纹状体和海马之间蛋白质组学结果的差异表明，在将来研究药物诱导的神经毒性时，有必要考虑大脑区域的差异和相互交织的信号通路，而不是仅仅关注单个核或分子。海马和纹状体之间还有其他一些明显不同的途径。结论我们的数据揭示了饮酒后大脑区域的差异，并表明了GR信号转导的角蛋白在酒精神经毒性中的关键作用。纹状体和海马之间蛋白质组学结果的差异表明，在将来研究药物诱导的神经毒性时，有必要考虑大脑区域的差异和相互交织的信号通路，而不是仅仅关注单个核或分子。海马和纹状体之间还有其他一些明显不同的途径。结论我们的数据揭示了饮酒后大脑区域的差异，并表明了GR信号转导的角蛋白在酒精神经毒性中的关键作用。纹状体和海马之间蛋白质组学结果的差异表明，在将来研究药物诱导的神经毒性时，有必要考虑大脑区域的差异和相互交织的信号通路，而不是仅仅关注单个核或分子。