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

    Author(s)
    Rose'Meyer, RB
    Mellick, AS
    Garnham, BG
    Harrison, GJ
    Massa, HM
    Griffiths, LR
    Griffith University Author(s)
    Harrison, Glenn J.
    Griffiths, Lyn
    Massa, Helen M.
    Rose'Meyer, Roselyn B.
    Garnham, Bronwyn G.
    Mellick, Albert S.
    Year published
    2003
    Metadata
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    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) a and ߮ 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 ...
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    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) a and ߮ 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-߬ following ovariectomy. No change was found in ADORA2B or ER-a. 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.
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    Journal Title
    Brain Research Protocols
    Volume
    11
    Publisher URI
    http://www.elsevier.com/wps/find/journaldescription.cws_home/622287/description#description
    DOI
    https://doi.org/10.1016/S1385-299X(02)00219-2
    Copyright Statement
    © 2003 Elsevier : Reproduced in accordance with the copyright policy of the publisher : This journal is available online - use hypertext links
    Subject
    Neurosciences
    Psychology
    Cognitive Sciences
    Publication URI
    http://hdl.handle.net/10072/6190
    Collection
    • Journal articles

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