Bowles Center for Alcohol Studies, The University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Carolina Institute for Developmental Disabilities, The University of North Carolina at Chapel Hill, Chapel Hill, NC, United States; UNC Neuroscience Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States; Department of Genetics, The University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Alcohol. 2023 Feb;106:1-9. doi: 10.1016/j.alcohol.2022.09.001. Epub 2022 Oct 4.
Alcohol exposure during the formation and closure of the neural tube, or neurulation (embryonic day [E] 8-10 in mice; ∼4th week of human pregnancy), perturbs development of midline brain structures and significantly disrupts gene expression in the rostroventral neural tube (RVNT). Previously, alcohol exposure during neurulation was found to alter gene pathways related to cell proliferation, p53 signaling, ribosome biogenesis, immune signaling, organogenesis, and cell migration 6 or 24 h after administration. Our current study expands upon this work by investigating short-term gene expression changes in the RVNT following a single binge-like alcohol exposure during neurulation. Female C57BL/6J mice were administered a single dose of 2.9 g/kg alcohol or vehicle on E9.0 to target mid-neurulation. The RVNTs of stage-matched embryos were collected 2 or 4 h after exposure and processed for RNA-seq. Functional profiling was performed with g:Profiler, as well as with the CiliaCarta and DisGeNet databases. Two hours following E9.0 alcohol exposure, 650 genes in the RVNT were differentially expressed. Functional enrichment analysis revealed that pathways related to cellular metabolism, gene expression, cell cycle, organogenesis, and Hedgehog signaling were down-regulated, and pathways related to cellular stress response, p53 signaling, and hypoxia were up-regulated by alcohol. Four hours after alcohol exposure, 225 genes were differentially expressed. Biological processes related to metabolism, RNA binding, ribosome biogenesis, and methylation were down-regulated, while protein localization and binding, autophagy, and intracellular signaling pathways were up-regulated. Two hours after alcohol exposure, the differentially expressed genes were associated with disease terms related to eye and craniofacial development and anoxia. These data provide further information regarding the biological functions targeted by alcohol exposure during neurulation in regions of the neural tube that give rise to alcohol-sensitive midline brain structures. Disruption of these gene pathways contributes to the craniofacial and brain malformations associated with prenatal alcohol exposure.
神经管（神经胚形成和闭合期，即小鼠 E8-10 天；人类妊娠第 4 周）形成和闭合期间暴露于酒精会扰乱中线脑结构的发育，并显著破坏头侧神经管（RVNT）中的基因表达。此前发现，神经胚形成期暴露于酒精会改变与细胞增殖、p53 信号转导、核糖体生物发生、免疫信号转导、器官发生和细胞迁移相关的基因途径，这些改变在给药后 6 或 24 小时出现。我们的研究通过在神经胚形成期单次 binge-like 酒精暴露后研究 RVNT 的短期基因表达变化，进一步扩展了这一工作。C57BL/6J 雌性小鼠在 E9.0 时接受 2.9 g/kg 酒精或载体单次给药，以靶向中神经胚形成期。暴露后 2 或 4 小时收集阶段匹配胚胎的 RVNTs ，并进行 RNA-seq 处理。使用 g:Profiler 以及 CiliaCarta 和 DisGeNet 数据库进行功能谱分析。E9.0 酒精暴露后 2 小时，RVNT 中有 650 个基因表达差异。功能富集分析显示，与细胞代谢、基因表达、细胞周期、器官发生和 Hedgehog 信号转导相关的途径下调，与细胞应激反应、p53 信号转导和缺氧相关的途径上调。E9.0 酒精暴露后 4 小时，有 225 个基因表达差异。与代谢、RNA 结合、核糖体生物发生和甲基化相关的生物学过程下调，而蛋白质定位和结合、自噬和细胞内信号通路上调。E9.0 酒精暴露后 2 小时，差异表达基因与与眼和颅面发育及缺氧相关的疾病术语相关。这些数据进一步提供了有关神经管中与酒精敏感中线脑结构相关区域的神经胚形成期酒精暴露靶向的生物学功能的信息。这些基因途径的破坏导致与产前酒精暴露相关的颅面和脑畸形。
