The measurement of adenosine and estrogen receptor expression in rat brains following ovariectomy using quantitative PCR analysis

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Title The measurement of adenosine and estrogen receptor expression in rat brains following ovariectomy using quantitative PCR analysis
Author Rose'Meyer, Roselyn; Mellick, Albert; Garnham, Bronwyn Gaye; Harrison, Glenn; Massa, Helen Maureen; Griffiths, Lyn
Journal Name Brain Research Protocols
Editor F G Wouterlood
Year Published 2003
Place of publication Netherlands
Publisher Elsevier BV
Abstract In our laboratory we have developed a quantitative-polymerase chain reaction (Q-PCR) strategy to examine the differential expression of adenosine receptor (ADOR), A1, A2A, A2B and A3, and estrogen receptors (ER) α and β. Brain and uterine mRNA were first used to optimise specific amplification conditions prior to SYBR Green I real time analysis of receptor subtype expression. SYBR Green I provided a convenient and sensitive means of examining specific PCR amplification product in real time, and allowed the generation of standard curves from which relative receptor abundance could be determined. Real time Q-PCR analysis was then performed, to examine changes in receptor expression levels in brains of adult female Wistar rats 3-month post ovariectomy. Comparison with sham-operated age-matched control rats demonstrated both comparative and absolute-copy number changes in receptor levels. Evaluation of both analytical methods investigated 18S rRNA as an internal reference for comparative gene expression analysis in the brain. The results of this study revealed preferential repression of ADORA2A (>4-fold down) and consistent (>2-fold) down-regulation of ADORA1, ADORA3, and ER-β, following ovariectomy. No change was found in ADORA2B or ER-α. Analysis of absolute copy number in this study revealed a correlation between receptor expression in response to ovariectomy, and relative receptor subtype abundance in the brain.
Peer Reviewed Yes
Published Yes
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Copyright Statement Copyright 2003 Elsevier : Reproduced in accordance with the copyright policy of the publisher : This journal is available online - use hypertext links
Volume 11
Page from 9
Page to 18
ISSN 1385-299X
Date Accessioned 2004-03-31
Date Available 2008-11-30T22:45:12Z
Language en_AU
Research Centre Griffith Health Institute; Molecular Basis of Disease; Heart Foundation Research Centre
Faculty Griffith Health Faculty
Subject Cardiology (incl. Cardiovascular Diseases)
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1

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