The Effect of Hypoxia on G Protein Coupled (Opioid) Receptor Gene Expression in Cortical B50 Neurons in Culture



Ibegbu, A O and Mullaney, I and Fyfe, Lorna and McBean, Douglas (2011) The Effect of Hypoxia on G Protein Coupled (Opioid) Receptor Gene Expression in Cortical B50 Neurons in Culture. British Journal of Pharmacology and Toxicology, 2 (2). pp. 75-83. ISSN 2044-2467

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Abstract

Hypoxia adversely affects cells and tissues, and neuronal cells in particular have been shown to be more susceptible to the injurious effects of hypoxia in which they may begin to die when oxygen supply is reduced or completely eliminated. Opioid receptor agonists have been shown to elicit several central nervous system effects, mediated via G protein-coupled receptors. The aim of this study was to study the effect of hypoxia on G protein coupled receptor gene expression using mu opioid receptor as a case study in cortical neuronal B50 cell lines in culture. The B50 cells were cultured in normoxia (21% O2; 5% CO2) and hypoxia (5% O2; 5% CO2), and were treated with opioid agonists to determine their effects on hypoxia-induced changes. Three opioid agonists {DAMGO(μ), DSLET(*) and ICI--199,441(6)}, were administered to the cells as treatment for 48 hours after 48 hours of initial culture for a total of 96 hours of culture in hypoxic conditions at concentrations of 10, 50 and 100 :M. The levels of G-protein coupled receptor (mu opioid) mRNAs were assessed using RT-PCR. The results showed that hypoxia induced morphological changes in B50 cells in hypoxia while the mu opioid RT-PCR mRNA levels showed no appreciable changes in normal, hypoxic and treated cells. The results show that B50 neuronal cells are susceptible to damage and injurious effects of hypoxia, as are most brain cells and the opioid agonist treatments showed there were no changes in the level of mu opioid receptor gene expression due to hypoxia or agonist treatment in neuronal B50 cells in culture.

Item Type: Article
Additional Information: The authors acknowledge Queen Margaret University, Edinburgh for the award of the Martlet research Scholarship and the Ahmadu Bello University Zaria-Nigeria for awarding the first author study fellowship to undertake this research studies. The authors would like to thank Promega Corporation for generously providing us with free samples and assay kits and reagents. Our special thanks go to Drs Paul Kelly and Linda Ferrington of the Centre for Neuroscience, University of Edinburgh for their help and guidance in RT-PCR technique. Our thanks go to Dr. Elizabeth Fashola-Stone, Technical Manager European collection of cell cultures (ECACC), for providing specialist and technical advice on the use of B50 cells
Uncontrolled Keywords: Hypoxia, G-protein coupled receptor, Opioid (mu) receptor, messenger RNA, reverse transcription-PCR, opioid agonist
Divisions: School of Health Sciences > Dietetics, Nutrition and Biological Sciences
Date Deposited: 27 Mar 2012 10:57
Last Modified: 19 Mar 2014 12:59
URI: http://eresearch.qmu.ac.uk/id/eprint/2727

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