Skip to main content

Analysis of Nrf2-Mediated Transcriptional Induction of Antioxidant Response in Early Embryos

  • Protocol
  • First Online:
Developmental Toxicology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 889))

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Giudice A, Montella M (2006) Activation of the Nrf2-ARE signaling pathway: a promising strategy in cancer prevention. Bioessays 28:169–181

    Article  PubMed  CAS  Google Scholar 

  2. Nguyen T, Nioi P, Pickett CB (2009) The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 284:13291–13295

    Article  PubMed  CAS  Google Scholar 

  3. Kensler TW, Wakabayashi N, Biswal S (2007) Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. Annu Rev Pharmacol Toxicol 47:89–116

    Article  PubMed  CAS  Google Scholar 

  4. Zhang DD (2006) Mechanistic studies of the Nrf2-Keap1 signaling pathway. Drug Metab Rev 38:769–789

    Article  PubMed  CAS  Google Scholar 

  5. Li WG, Kong AN (2009) Molecular mechanisms of Nrf2-mediated antioxidant response. Mol Carcinog 48:91–104

    Article  PubMed  CAS  Google Scholar 

  6. Cho HY, Jedlicka AE, Reddy SP et al (2002) Role of NRF2 in protection against hyperoxic lung injury in mice. Am J Respir Cell Mol Biol 26:175–182

    PubMed  CAS  Google Scholar 

  7. Lee JM, Shih AY, Murphy TH et al (2003) NF-E2-related factor-2 mediates neuroprotection against mitochondrial complex I inhibitors and increased concentrations of intracellular calcium in primary cortical neurons. J Biol Chem 278:37948–37956

    Article  PubMed  CAS  Google Scholar 

  8. Lee JM, Calkins MJ, Chan K et al (2003) Identification of the NF-E2-related factor-2-dependent genes conferring protection against oxidative stress in primary cortical astrocytes using oligonucleotide microarray analysis. J Biol Chem 278:12029–12038

    Article  PubMed  CAS  Google Scholar 

  9. Danilov CA, Chandirasekaran K, Racz J et al (2009) Sulforaphane protects astrocytes against oxidative stress and delayed death caused by oxygen and glucose deprivation. Glia 57:645–656

    Article  PubMed  Google Scholar 

  10. Zhang JD, Hung AC, Ng PY et al (2009) PKC delta mediates Nrf2-dependent protection of neuronal cells from NO-induced apoptosis. Biochem Biophys Res Commun 386:750–756

    Article  PubMed  CAS  Google Scholar 

  11. Yan D, Dong J, Sulik KK, Chen S-Y (2010) Induction of the Nrf2-driven antioxidant response by tert-butylhydroquinone prevents ethanol-induced apoptosis in cranial neural crest cells. Biochem Pharmacol 80:144–149

    Article  PubMed  CAS  Google Scholar 

  12. Dong J, Sulik KK, Chen SY (2008) Nrf2-mediated transcriptional induction of antioxidant response in mouse embryos exposed to ethanol in vivo: implications for the prevention of fetal alcohol spectrum disorders. Antioxid Redox Signal 10:2023–2033

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The author would like to thank Drs. Jian Dong, Haijing Sun, and Ms Jie Liu for technical assistance. This work is supported by NIH grants AA017446 and AA013908.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Shao-yu Chen Ph.D. .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Chen, Sy. (2012). Analysis of Nrf2-Mediated Transcriptional Induction of Antioxidant Response in Early Embryos. In: Harris, C., Hansen, J. (eds) Developmental Toxicology. Methods in Molecular Biology, vol 889. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-867-2_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-867-2_17

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-866-5

  • Online ISBN: 978-1-61779-867-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics