Analysis of Nrf2-mediated transcriptional induction of antioxidant response in early embryos.
Nrf2 is a transcription factor that regulates the induction of an expansive set of antioxidant proteins that act in synergy to remove reactive oxygen species (ROS). We have recently discovered that Nrf2 signaling is involved in the induction of antioxidant response in ethanol-exposed embryos. We have also demonstrated that chemically induced transcriptional activation of Nrf2 and subsequent induction of a broad spectrum of antioxidant proteins can act as an endogenous protective system against ethanol-induced oxidative stress and apoptosis in early mouse embryos. This chapter describes the methods used to analyze Nrf2-mediated transcriptional induction of antioxidant response in mouse embryos. Data which have been generated by using these methods have also been included to provide examples of their usefulness in evaluating Nrf2-mediated antioxidant response in early embryos.
Department of Cancer Biology and Pharmacology, University of Illinois College of Medicine at Peoria-UICOM-P, Peoria, IL, USA. sychen@uic.edu
Shao-yu
SY
Chen
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D004195:Disease Models, Animal; D024202:Electrophoretic Mobility Shift Assay; D004622:Embryo, Mammalian; D057075:Enzyme Assays; D000431:Ethanol / Q000633:toxicity*; D005260:Female; D063647:Fetal Alcohol Spectrum Disorders; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D008431:Maternal-Fetal Exchange; D051379:Mice; D008810:Mice, Inbred C57BL; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D011247:Pregnancy; D011485:Protein Binding; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020218:Response Elements; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D000099156:Superoxide Dismutase 2
D000975:Antioxidants; D002374:Catalase; D002457:Cell Extracts; D000431:Ethanol; D051267:NF-E2-Related Factor 2; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D000099156:Superoxide Dismutase 2; C495636:Nfe2l2 protein, mouse; C000606291:Sod1 protein, mouse