Housekeepers for accurate transcript expression analysis in Alzheimer's disease autopsy brain tissue

Gebhardt, Florian M., Scott, Heather A. and Dodd, Peter R. (2010) Housekeepers for accurate transcript expression analysis in Alzheimer's disease autopsy brain tissue. Alzheimer's and Dementia, 6 6: 465-474. doi:10.1016/j.jalz.2009.11.002

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Author Gebhardt, Florian M.
Scott, Heather A.
Dodd, Peter R.
Title Housekeepers for accurate transcript expression analysis in Alzheimer's disease autopsy brain tissue
Formatted title
Housekeepers for accurate transcript expression analysis in Alzheimer’s disease autopsy brain tissue
Journal name Alzheimer's and Dementia   Check publisher's open access policy
ISSN 1552-5260
Publication date 2010-11
Sub-type Article (original research)
DOI 10.1016/j.jalz.2009.11.002
Volume 6
Issue 6
Start page 465
End page 474
Total pages 10
Editor Zaven Khachaturian
Place of publication New York, NY, U.S.A.
Publisher Elsevier
Collection year 2011
Language eng
Formatted abstract
Background Quantitative real-time reverse transcription–polymerase chain reaction (qRT-PCR) is a popular technique for mRNA expression studies. Normalization to an endogenous reference transcript (housekeeper) is widely used to correct for differences in loading and RNA quality. Alzheimer's disease (AD) alters brain metabolism. The stability of housekeeper transcript expression must be carefully validated.

Methods qRT-PCR was used to assess eight putative housekeeper transcripts in four brain regions from 15 control, 12 AD, and 10 AD/Lewy body disease (LBD) cases.

Results RNA quality is lower in AD and AD/LBD than in controls. Frequently used housekeepers such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin had lower overall expression in AD and AD/LBD cases than in controls. RPL13 and 18S were the most stably expressed housekeepers tested. Synaptophysin and glial fibrillary acidic protein were used to evaluate normalized quantification. By using different housekeepers we confirmed that synaptophysin expression was down-regulated in AD cases, whereas glial fibrillary acidic protein expression was increased.

Conclusions Among all candidates tested, RPL13 was the best housekeeper for qRT-PCR studies in autopsy brain tissue samples from controls and AD cases. RNA quality should be assessed and data normalized on this index as well.
© 2010 The Alzheimer’s Association. All rights reserved.
Keyword Quantitative real-time PCR
RNA integrity number
Postmortem interval
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Official 2011 Collection
School of Chemistry and Molecular Biosciences
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Citation counts: TR Web of Science Citation Count  Cited 13 times in Thomson Reuters Web of Science Article | Citations
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Created: Tue, 09 Nov 2010, 09:21:11 EST by Jon Swabey on behalf of School of Chemistry & Molecular Biosciences