Smirnova Lena, Seiler Andrea E M, Luch Andreas
Current address: Center for Alternatives to Animal Testing (CAAT), Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland.
Department of Experimental Toxicology and Center for Documentation and Evaluation of Alternatives to Animal Experiments (ZEBET), German Federal Institute for Risk Assessment (BfR), Berlin, Germany.
Curr Protoc Toxicol. 2015 May 4;64:20.9.1-20.9.22. doi: 10.1002/0471140856.tx2009s64.
microRNAs (miRNAs) are small non-coding RNA molecules functioning as post-transcriptional regulators of gene expression. miRNAs play a significant role in organism development, regulating developmental timing, cell differentiation, and specification. In the developing brain, miRNAs regulate neural stem cell differentiation, lineage specification, synaptogenesis, and brain morphogenesis. Temporal and spatial specificity of miRNA expression make them an attractive marker to study cellular responses to toxicant exposure. Neural differentiation of murine embryonic stem cells (mESCs) has been established as an alternative method to study developmental neurotoxicity (DNT) in vitro. This unit will describe a method for miRNA profiling (miRNomics) as a molecular end point to study developmental neurotoxicity. A protocol for neural differentiation of mESC will be described as a cellular model for DNT testing. The miRNomics protocol is versatile and can be used with other DNT cellular systems such as primary cultures, human embryonic stem cells (hESCs), or induced pluripotent stem cells (iPSCs).
微小RNA（miRNA）是一类小的非编码RNA分子，作为基因表达的转录后调节因子发挥作用。miRNA在生物体发育中起着重要作用，调节发育时间、细胞分化和细胞特化。在发育中的大脑中，miRNA调节神经干细胞分化、谱系特化、突触形成和脑形态发生。miRNA表达的时空特异性使其成为研究细胞对毒物暴露反应的有吸引力的标志物。小鼠胚胎干细胞（mESC）的神经分化已被确立为一种在体外研究发育性神经毒性（DNT）的替代方法。本单元将描述一种作为研究发育性神经毒性分子终点的miRNA谱分析（miRNomics）方法。将描述mESC神经分化的方案，作为DNT测试的细胞模型。miRNomics方案具有通用性，可与其他DNT细胞系统一起使用，如原代培养物、人类胚胎干细胞（hESC）或诱导多能干细胞（iPSC）。
