Astrocyte-specific transcriptome analysis using the ALDH1L1 bacTRAP mouse reveals novel biomarkers of astrogliosis in response to neurotoxicity
Astrocyte-specific transcriptome analysis using the ALDH1L1 bacTRAP mouse reveals novel biomarkers of astrogliosis in response to neurotoxicity
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DOI：
10.1111/jnc.14800
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发表时间：
2019-07-11
影响因子：
4.7
通讯作者：
O'Callaghan，James P.
中科院分区：
文献类型：
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作者：
Michalovicz，Lindsay T.;Kelly，Kimberly A.;O'Callaghan，James P.
神经毒理学因无法预测毒物引起的损伤的区域和细胞目标而受到阻碍。评估星形胶质细胞增生可以克服这个问题，因为反应性星形胶质细胞突出了毒物引起的损伤的位置。虽然神经胶质原纤维酸性蛋白表达增强是星形胶质细胞增生的标志，但尚未发现其他生物标志物。然而，细菌人工染色体翻译核糖体亲和纯化（bacTRAP）技术可以表征特定细胞类型的主动翻译转录组；利用这一技术在乙醛脱氢酶1家族成员L1（ALDH1L1）bacTRAP小鼠中可以识别星形胶质细胞中选择性表达的基因。本研究的目的是使用 ALDH1L1 bacTRAP 小鼠来表征神经毒性诱导的星形胶质细胞增生的其他生物标志物。已知的多巴胺能神经毒剂 1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP；12.5 mg/kg s.c.）用于诱导星形胶质细胞增生。暴露后 12、24 和 48 小时获取纹状体组织，以分离主动翻译的 RNA。随后，通过微阵列分析 MPTP 诱导的 RNA 库变化，并鉴定出 184 个具有统计学意义的差异表达基因。通过基因本体、通路和共表达网络分析对数据集进行了询问，确定了新基因以及具有已知免疫和炎症功能的基因。通过这些分析，我们找到了与反应性星形胶质细胞相关的几个基因。其中，TIMP1 和 miR-147 被确定为候选生物标志物，因为它们在 MPTP 和三甲基锡暴露后表达显着增加。因此，我们已经证明 bacTRAP 可用于识别星形胶质细胞增生的新生物标志物，并有助于表征有毒物质引起的星形胶质细胞表型。开放科学徽章本文已获得“开放材料”徽章，因为它提供了在手稿中重现研究的所有相关信息。本文的完整开放科学披露表格可以在文章末尾找到。有关开放实践徽章的更多信息，请访问 https://cos.io/our-services/open-science-badges/。本期封面图片：doi: 10.1111/jnc.14518。
Neurotoxicology is hampered by the inability to predict regional and cellular targets of toxicant‐induced damage. Evaluating astrogliosis overcomes this problem because reactive astrocytes highlight the location of toxicant‐induced damage. While enhanced expression of glial fibrillary acidic protein is a hallmark of astrogliosis, few other biomarkers have been identified. However, bacterial artificial chromosome ‐ translating ribosome affinity purification (bacTRAP) technology allows for characterization of the actively translating transcriptome of a particular cell type; use of this technology in aldehyde dehydrogenase 1 family member L1 (ALDH1L1) bacTRAP mice can identify genes selectively expressed in astrocytes. The aim of this study was to characterize additional biomarkers of neurotoxicity‐induced astrogliosis using ALDH1L1 bacTRAP mice. The known dopaminergic neurotoxicant 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP; 12.5 mg/kg s.c.) was used to induce astrogliosis. Striatal tissue was obtained 12, 24, and 48 h following exposure for the isolation of actively translating RNA. Subsequently, MPTP‐induced changes in this RNA pool were analyzed by microarray and 184 statistically significant, differentially expressed genes were identified. The dataset was interrogated by gene ontology, pathway, and co‐expression network analyses, which identified novel genes, as well as those with known immune and inflammatory functions. Using these analyses, we were directed to several genes associated with reactive astrocytes. Of these, TIMP1 and miR‐147 were identified as candidate biomarkers because of their robust increased expression following both MPTP and trimethyl tin exposures. Thus, we have demonstrated that bacTRAP can be used to identify new biomarkers of astrogliosis and aid in the characterization of astrocyte phenotypes induced by toxicant exposures.Open Science BadgesThis article has received a badge for *Open Materials* because it provided all relevant information to reproduce the study in the manuscript. The complete Open Science Disclosure form for this article can be found at the end of the article. More information about the Open Practices badges can be found at https://cos.io/our-services/open-science-badges/.Cover Image for this issue: doi: 10.1111/jnc.14518.