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Monitoring of cell oxygenation and responses to metabolic stimulation by intracellular oxygen sensing technique
Alternative Title
The Determinants of Oxygen Gradients in Respiring Samples and their Impact on Cellular Responses to Hypoxia
Date Issued
2010
Date Available
2013-11-29T09:16:52Z
Abstract
Quenched-phosphorescence oxygen (O2) sensing technique allows non-invasive, real-time monitoring of both intra- and extracellular O2 concentration in respiring samples. Using this technique we investigated O2 gradients in populations of neurosecretory PC12 cells cultured in 96-well plates and exposed to graded hypoxia at rest and upon metabolic stimulation. Under high atmospheric O2 (10–21%) the respiration of resting cells dictated that local O2 was moderately reduced, and at a certain threshold (6% in galactose medium) cell layer became practically anoxic. Furthermore, cell stimulation triggered a major redistribution of O2 and a prominent ‘hypoxic overshoot’ mediated by diffusion. The deep, prolonged cell deoxygenation upon stimulation was matched by an increase in nuclear HIF-1α levels. In the presence of nitric oxide the hypoxic overshoot was truncated and HIF-1α stabilization inhibited. Thus, the main determinants which impact upon cellular O2 levels and oxygen-sensitive signaling pathways are the atmospheric O2, sample geometry, cell density, respiration rate and its dynamics. Changes in any of these parameters can significantly alter the O2 levels experienced by the cells and the subsequently activated signaling pathways. This technique, which provides simple and reliable monitoring of cell oxygenation, is therefore important for hypoxia research, metabolic studies and experiments with respiring cells.
Other Sponsorship
Science Foundation Ireland, grant 07/IN.1/B1804 and Enterprise Ireland, grant EI CFTD/07/124
Type of Material
Journal Article
Publisher
The Royal Society of Chemistry
Journal
Integrative Biology
Volume
2
Issue
9
Copyright (Published Version)
2010 The Royal Society of Chemistry
Language
English
Status of Item
Peer reviewed
This item is made available under a Creative Commons License
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