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Cited by 6Adenosine and Purinergic SignalingEstrogen and related hormone effectsMolecular Biology Techniques and Applications

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

Rose’Meyer, Roselyn B, Mellick, Albert S, Garnham, Bronwyn G, Harrison, Glenn J, Massa, Helen M, Griffiths, Lyn R·2003·Brain Research Protocols
DOI 10.1016/s1385-299x(02)00219-2
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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), A(1), A(2A), A(2B) and A(3), and estrogen receptors (ER) alpha and beta. 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 ADORA(2A) (>4-fold down) and consistent (>2-fold) down-regulation of ADORA(1), ADORA(3), and ER-beta, following ovariectomy. No change was found in ADORA(2B) or ER-alpha. 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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  • Following quiescence in primates, hormones transiently have different effects on the circadian system than those observed in species with low-level steroidogenesis throughout the juvenile period.

    “Since long-term hormone withdrawal is known to alter hormone receptor levels [122, 161], it seems more likely that following quiescence in primates, hormones transiently have different effects on the circadian system than those observed in species with low-level steroidogenesis throughout the juvenile period.”

    From The neuroendocrine control of the circadian system: Adolescent chronotype (2012) · cites this paper as [161]

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Authors
Rose’Meyer, R. B., Mellick, A. S., Garnham, B. G., Harrison, G. J., Massa, H. M., & Griffiths, L. R.
Affiliations
Griffith University (AU)
Corresponding author
Rose’Meyer, Roselyn B
ORCID
Venue
Brain Research Protocols
Publisher
Elsevier BV
Published
2003-03
Volume
11
Issue
1
Pages
9-18
ISSN
1385-299X (ISSN-L)
DOI
10.1016/s1385-299x(02)00219-2
OpenAlex
W1494387119
Cited by
4
References
25
Citations by year
2022: 1 · 2021: 1 · 2012: 1
FWCI
0.00
Type
journal article
Language
en
Open access
closed
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Brain Research Protocols (publishedVersion)
  • Institute of Health and Biomedical Innovation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Adenosine and Purinergic Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
Keywords
adenosine receptors, estrogen receptor, ovariectomy, brain gene expression, adenosine A2A receptor, adenosine A1 receptor, A3 adenosine receptor, adenosine A2B receptor, estrogen receptor alpha, estrogen receptor beta, real-time PCR, SYBR Green I, female rats, menopause

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