Document Detail

Monitoring of cell oxygenation and responses to metabolic stimulation by intracellular oxygen sensing technique.
MedLine Citation:
PMID:  20657921     Owner:  NLM     Status:  MEDLINE    
Quenched-phosphorescence oxygen (O(2)) sensing technique allows non-invasive, real-time monitoring of both intra- and extracellular O(2) concentration in respiring samples. Using this technique we investigated O(2) 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 O(2) (10-21%) the respiration of resting cells dictated that local O(2) 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 O(2) and a prominent 'hypoxic overshoot' mediated by diffusion. The deep, prolonged cell deoxygenation upon stimulation was matched by an increase in nuclear HIF-1alpha levels. In the presence of nitric oxide the hypoxic overshoot was truncated and HIF-1alpha stabilization inhibited. Thus, the main determinants which impact upon cellular O(2) levels and oxygen-sensitive signaling pathways are the atmospheric O(2), sample geometry, cell density, respiration rate and its dynamics. Changes in any of these parameters can significantly alter the O(2) 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.
Alexander V Zhdanov; Vladimir I Ogurtsov; Cormac T Taylor; Dmitri B Papkovsky
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-07-26
Journal Detail:
Title:  Integrative biology : quantitative biosciences from nano to macro     Volume:  2     ISSN:  1757-9708     ISO Abbreviation:  Integr Biol (Camb)     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-09-02     Completed Date:  2010-12-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101478378     Medline TA:  Integr Biol (Camb)     Country:  England    
Other Details:
Languages:  eng     Pagination:  443-51     Citation Subset:  IM    
Biochemistry Department, University College Cork, Cork, Ireland.
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MeSH Terms
Cell Count
Cell Hypoxia
Cell Respiration / drug effects
Egtazic Acid / pharmacology
Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
Models, Biological
Oxygen / metabolism*
PC12 Cells
Signal Transduction
Reg. No./Substance:
0/Hif1a protein, rat; 0/Hypoxia-Inducible Factor 1, alpha Subunit; 67-42-5/Egtazic Acid; 7782-44-7/Oxygen

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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