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In Vitro Neurotoxicology pp 205–214Cite as

Assessment of Glutathione Homeostasis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 758))

Abstract

The tripeptide glutathione (γ-glutamylcysteinylglycine; GSH) is the most abundant antioxidant thiol in the brain. GSH plays a critical role in protecting brain cells from oxidative stress and xenobiotics, as well as maintaining the thiol redox state. High levels of GSH are present in the central nervous system, particularly in astrocytes. GSH is synthesized into two enzymatic steps, the first, and the rate-limiting one, is catalyzed by glutamate–cysteine ligase (GCL) to form a dipeptide which is then converted to GSH by GSH synthetase. In this chapter, we describe assays for the measurements of GSH levels and GCL activity. The first spectrophotometric assay is based on the affinity of 2,3-naphthalenedicarboxaldehyde (NDA) for GSH. In the second assay, GSH levels are measured after being derivatized using the fluorescent thiol reactive compound monobromobimane (MBB) and are detected by high-performance liquid chromatography (HPLC). The third assay allows the assessment of GCL activity, also by HPLC.

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Author information

Authors and Affiliations

  1. Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA, USA

    Lucio G. Costa

  2. Department of Human Anatomy, Pharmacology, and Forensic Science, University of Parma Medical School, Parma, Italy

    Lucio G. Costa

Authors
  1. Gennaro Giordano
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  2. Collin C. White
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  3. Lucio G. Costa
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Corresponding author

Correspondence to Lucio G. Costa .

Editor information

Editors and Affiliations

  1. , Department of Environmental and, University of Washington, Roosevelt Way NE 4225, Seattle, 98105, Washington, USA

    Lucio G. Costa

  2. , Department of Environmental and, University of Washington, Roosevelt Way NE 4225, Seattle, 98105, Washington, USA

    Gennaro Giordano

  3. Jesse Brown VA Medical Center, Department of Psychiatry, University of Illinois at Chicago, South Damen Avenue 820, Seattle, 60612, Illinois, USA

    Marina Guizzetti

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Giordano, G., White, C.C., Costa, L.G. (2011). Assessment of Glutathione Homeostasis. In: Costa, L., Giordano, G., Guizzetti, M. (eds) In Vitro Neurotoxicology. Methods in Molecular Biology, vol 758. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-170-3_14

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  • DOI: https://doi.org/10.1007/978-1-61779-170-3_14

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-169-7

  • Online ISBN: 978-1-61779-170-3

  • eBook Packages: Springer Protocols

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