Evaluation of fluorescent dyes for measuring intracellular glutathione content in primary cultures of human neurons and neuroblastoma SH‐SY5Y

@article{Sebasti2003EvaluationOF,
  title={Evaluation of fluorescent dyes for measuring intracellular glutathione content in primary cultures of human neurons and neuroblastoma SH‐SY5Y},
  author={Jordi Ponce Sebasti{\`a} and Rosa M. Crist{\`o}fol and Manuela Mart{\'i}n and Eduard Rodr{\'i}guez-Farr{\'e} and Coral Sanfeliu},
  journal={Cytometry Part A},
  year={2003},
  volume={51A}
}
Reduced glutathione (GSH) protects cells against oxidative injury and maintains a range of vital functions. To study GSH content in human neuronal cell cultures, thiol‐sensitive fluorescent techniques requiring a small number of cells may be of great value, but their GSH specificity has not been established in these cells. 
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110 Citations

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References

SHOWING 1-10 OF 29 REFERENCES

Characterization of Hydrogen Peroxide Toxicity in Cultured Rat Forebrain Neurons

TLDR
It is demonstrated that H2O2 is a potent and effective neurotoxin that produces oxidative stress, as well as apoptotic neuronal death, in cultured rat forebrain neurons.

Glutathione levels in primary glial cultures: Monochlorobimane provides evidence of cell type‐specific distribution

TLDR
It seems that owing to the greater intracellular concentration of reactive oxygen and nitrogen species to which microglia are subjected, especially under conditions of inflammation, this cell type is fortified with higher GSH levels as a means to combat oxidative and nitrosative stress.

Evaluation of glutathione‐sensitive fluorescent dyes in cortical culture

TLDR
Neurons experienced a dramatic decline in GSH levels relative to astrocytes between 5–6 DIV, as shown by a loss of neuronal staining with CMAC, CMAC‐blue and MCB, indicating specificity for GSH.

Direct evidence for glutathione as mediator of apoptosis in neuronal cells.

Glutathione depletion and neuronal cell death: the role of reactive oxygen intermediates and mitochondrial function

Synthesis of the Antioxidant Glutathione in Neurons: Supply by Astrocytes of CysGly as Precursor for Neuronal Glutathione

TLDR
The data suggest the following metabolic interaction in glutathione metabolism of brain cells: the ectoenzyme γ-glutamyl transpeptidase uses as substrate the glutATHione released by astrocytes to generate the dipeptide CysGly that is subsequently used by neurons as precursor for glutathion synthesis.

Quantitative analysis of cellular glutathione by flow cytometry utilizing monochlorobimane: some applications to radiation and drug resistance in vitro and in vivo.

TLDR
Heterogeneity in populational GSH levels was observed under certain circumstances, such as release from hypoxia, and the significance of this heterogeneity is discussed in regard to the induction of gene amplification and drug resistance by transient Hypoxia.

Evaluation of methods for measuring cellular glutathione content using flow cytometry.

TLDR
It is recommended that flow cytometric GSH assays should routinely include a sample of cells that have been depleted of GSH in order to determine the extent of background labeling, and monobromobimane is the agent of choice.

Metabolism and functions of glutathione in brain

Glutathione metabolism in brain

TLDR
The main focus of this short review is recent results on glutathione metabolism of brain astrocytes and neurons in culture, which show that these two types of cell prefer different extracellular precursors for glutATHione.