microRNA Profiling as Tool for Developmental Neurotoxicity Testing (DNT).
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).
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.
Lena
L
Smirnova
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.
Andrea E M
AEM
Seiler
Department of Chemicals and Products Safety, German Federal Institute for Risk Assessment (BfR), Berlin, Germany.
Andreas
A
Luch
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods* / Q000592:standards
D035683:MicroRNAs
embryonic stem cells; in vitro developmental neurotoxicity; miRNA; miRNomics