Abstract |
Oxidative stress is associated with the pathology of acute and chronic neurodegenerative disease. We have cloned a human neuroglobin (Nb) construct and over-expressed this protein in cultured human neuronal cells to assess whether Nb ameliorates the cellular response to experimental hypoxia-reoxygenation (H/R) injury. Parental cells transfected with a blank (pDEST40) vector responded to H/R injury with a significant decrease in cellular ATP at 5 and 24 h after insult. This was coupled with increases in the cytosolic Ca(2+), and the transition metals iron (Fe), copper (Cu), and zinc (Zn) within the cell body, as monitored simultaneously using X-ray fluorescence microprobe imaging. Parental cell viability decreased over the same time period with a approximately 4 to 5-fold increase in cell death (maximum approximately 25%) matched by an increase in caspase 3/7 activation (peaking at a 15-fold increase after 24 h) and condensation of beta-actin along axonal processes. Over-expression of Nb inhibited ATP loss and except for significant decreases in the sulfur (S), chlorine (Cl), potassium (K) and Ca(2+) contents, maintained cellular ion homeostasis after H/R insult. This resulted in increased cell viability, significantly diminished caspase activation and maintenance of the beta-actin cytoskeletal structure and receptor-mediated endocytosis. These data indicate that bolstering the cellular content of Nb inhibits neuronal cell dysfunction promoted by H/R insult through multiple protective actions including: (i) maintenance of cellular bioenergetics; (ii) inhibition of Ca(2+) influx; (iii) a reduction in cellular uptake of Fe, Cu and Zn at the expense of S, Cl and K; and (iv) an enhancement of cell viability through inhibiting necrosis and apoptosis.
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Authors | Thi Thuy Hong Duong, Paul Kenneth Witting, Shane Tony Antao, Sarah Nicole Parry, Marina Kennerson, Barry Lai, Stefan Vogt, Peter Andrew Lay, Hugh Hamlyn Harris |
Journal | Journal of neurochemistry
(J Neurochem)
Vol. 108
Issue 5
Pg. 1143-54
(Mar 2009)
ISSN: 1471-4159 [Electronic] England |
PMID | 19154338
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)
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Chemical References |
- Cytoskeletal Proteins
- Nerve Tissue Proteins
- Neuroglobin
- Copper
- Adenosine Triphosphate
- Globins
- Iron
- Caspases
- Zinc
- Oxygen
- Calcium
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Topics |
- Adenosine Triphosphate
(metabolism)
- Analysis of Variance
- Calcium
(metabolism)
- Caspases
(metabolism)
- Cell Differentiation
- Cell Hypoxia
(drug effects, physiology)
- Cell Line, Tumor
- Copper
(metabolism)
- Cytoskeletal Proteins
(metabolism)
- Endocytosis
- Flow Cytometry
(methods)
- Globins
(genetics, metabolism)
- Humans
- Iron
(metabolism)
- Nerve Tissue Proteins
(genetics, metabolism)
- Neuroblastoma
- Neuroglobin
- Neurons
(drug effects, physiology)
- Oxygen
(pharmacology)
- Spectrometry, X-Ray Emission
(methods)
- Time Factors
- Transfection
(methods)
- Zinc
(metabolism)
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