Table_2_Ethanol-induced transcriptional and translational changes in Aldh1l1-Egfp/Rpl10a cortical astrocyte cultures.XLSX

The role astrocytes play in brain development and function has garnered greater attention as the diversity of roles they are involved in has become apparent. We have previously shown that ethanol-exposed astrocytes alter neuronal neurite outgrowth in an in vitro co-culture system and that ethanol alters the astrocyte-produced extracellular matrix (ECM) in vitro, with similar alterations in vivo. In this study, we utilized the translating ribosome affinity purification (TRAP) procedure in Aldh1l1-EGFP/Rpl10a transgenic mouse primary cortical astrocyte cultures to transcriptionally and translationally profile the astrocyte response to ethanol. We found a large number of differences between the total RNA pool and the translating RNA pool, indicating that the transcriptional state of astrocytes may not always reflect the translational state of astrocytes. In addition, there was a considerable overlap between ethanol-dysregulated genes in the total RNA pool and the translating RNA pool. Comparisons to published datasets indicate the in vitro model used here is most similar to PD1 or PD7 in vivo cortical astrocytes, and the ethanol-regulated genes showed a significant overlap with models of chronic ethanol exposure in astrocytes, a model of third-trimester ethanol exposure in the hippocampus and cerebellum, and an acute model of ethanol exposure in the hippocampus. These findings will further our understanding of the effects of ethanol on astrocyte gene expression and protein translation and how these changes may alter brain development and support the use of in vitro astrocyte cultures as models of neonatal astrocytes.

星形胶质细胞（astrocytes）在大脑发育与功能中所发挥的作用，随着其参与调控的功能多样性逐渐被阐明，愈发受到研究界的广泛关注。我们此前的研究证实，经乙醇暴露的星形胶质细胞可在体外共培养体系中改变神经元的神经突生长；同时乙醇会在体外环境中改变星形胶质细胞分泌的细胞外基质（extracellular matrix, ECM），该类改变在体内也存在相似表现。本研究利用Aldh1l1-EGFP/Rpl10a转基因小鼠原代皮层星形胶质细胞培养物中的翻译核糖体亲和纯化（translating ribosome affinity purification, TRAP）技术，对星形胶质细胞应答乙醇刺激的转录组与翻译组进行了谱分析。研究发现，总RNA池与翻译RNA池之间存在大量表达差异，这提示星形胶质细胞的转录组状态或许并不总能反映其翻译组的实际状态。此外，总RNA池与翻译RNA池中受乙醇调控异常的基因存在相当程度的重叠。通过与已发表数据集进行比对，本研究使用的体外模型与体内PD1或PD7皮层星形胶质细胞最为相似；且乙醇调控基因与星形胶质细胞慢性乙醇暴露模型、海马及小脑第三孕期乙醇暴露模型，以及海马急性乙醇暴露模型的差异基因谱均存在显著重叠。本研究结果将进一步深化我们对乙醇如何影响星形胶质细胞基因表达与蛋白质翻译，以及这些改变如何调控大脑发育的理解，并支持将体外星形胶质细胞培养作为新生儿星形胶质细胞研究模型的应用价值。