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Validation of endog...
Validation of endogenous controls for quantitative gene expression analysis : Application on brain cortices of human chronic alcoholics
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- Johansson, Sofia (author)
- Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
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- Fuchs, Andrea (author)
- Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
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- Ökvist, Anna (author)
- Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
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- Karimi, Mohsen (author)
- Karolinska Institutet
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- Harper, Clive (author)
- Discipline of Pathology, University of Sydney, NSW Australia
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- Garrick, Therese (author)
- Discipline of Pathology, University of Sydney, NSW Australia
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- Sheedy, Donna (author)
- Discipline of Pathology, University of Sydney, NSW Australia
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- Hurd, Yasmin (author)
- Departments of Psychiatry and Pharmacology and Biological Chemistry at Mount Sinai School of Medicine, New York, USA
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- Bakalkin, Georgy (author)
- Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden,Molecular neuropsychopharmacology
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- Ekström, Tomas J. (author)
- Karolinska Institutet
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(creator_code:org_t)
- Elsevier BV, 2007
- 2007
- English.
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In: Brain Research. - : Elsevier BV. - 0006-8993 .- 1872-6240. ; 1132:1, s. 20-8
- Related links:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
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- Real-time PCR is frequently used for gene expression quantification due to its methodological sensitivity and reproducibility. The gene expression is quantified by normalization to one or more reference genes, usually beta-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPD) or to ribosomal RNA (18S). However, different environmental or pathological conditions might also influence the expression of normalizing genes, which could severely skew the interpretation of quantitative results. This study evaluates whether 16 genes frequently used as endogenous controls in expression studies, can serve as such for comparison of human brain tissues of chronic alcoholics and control subjects. The prefrontal and motor cortices that are affected differently by chronic alcohol consumption were analyzed. The reference genes that have no or small differences in expression in alcoholics and control subjects, were found to be specific for each region: beta-actin (ACTB) and ribosomal large P0 (RPLP0) for the prefrontal cortex while importin 8 (IPO8) and RNA polymerase II (POLR2A) for the motor cortex. Four out of sixteen analyzed genes demonstrated significant differences in expression between alcoholics and controls: phosphoglycerate kinase (PGK1), hypoxanthine phosphoribosyl transferase (HPRT1) and peptidylprolyl isomerase A (PPIA) in the motor cortex and beta-2-microglobulin (B2M) in the prefrontal cortex. Our study demonstrates the importance of validation of endogenous control genes prior to real-time PCR analysis of human brain tissues. Prescribed and non-prescribed drugs, pathological or environmental conditions along with alcohol abuse may differentially influence expression of reference genes.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Keyword
- Reference gene
- Frontal
- Motor
- geNORM
- Real-time PCR
- LDA
- Molecular biology
- Molekylärbiologi
- Biology with specialization in Molecular Biology
- Biologi med inriktning mot molekylärbiologi
- Molecular Biology
- Molekylärbiologi
- Neuroscience
- Neurovetenskap
Publication and Content Type
- ref (subject category)
- art (subject category)
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To the university's database
- By the author/editor
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Johansson, Sofia
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Fuchs, Andrea
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Ökvist, Anna
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Karimi, Mohsen
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Harper, Clive
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Garrick, Therese
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show more...
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Sheedy, Donna
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Hurd, Yasmin
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Bakalkin, Georgy
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Ekström, Tomas J ...
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show less...
- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Cell and Molecul ...
- Articles in the publication
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Brain Research
- By the university
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Uppsala University
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Karolinska Institutet