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Obtaining High Quality RNA from Single Cell Populations in Human Postmortem Brain Tissue
Published on: August 6, 2009
Selection of reference gene expression in a schizophrenia brain cohort
Cynthia Shannon Weickert1, Donna Sheedy, Debora A Rothmond
1Schizophrenia Research Institute, Sydney, New South Wales, Australia. c.weickert@powmri.edu.au
The Australian and New Zealand Journal of Psychiatry
|January 16, 2010
Summary
This study found that while brain tissue pH and agonal state impact RNA quality in schizophrenia research, well-matched cohorts enable reliable detection of gene expression differences. Careful consideration of factors is key for human brain studies.
Area of Science:
- Neuroscience
- Genetics
- Biochemistry
Background:
- Establishing well-characterized human brain cohorts is crucial for postmortem research on schizophrenia's neuropathological basis.
- Specimen characteristics can influence RNA quality and gene expression analysis in postmortem brain tissue.
Purpose of the Study:
- To determine if specimen characteristics (diagnosis, age, PMI, pH, agonal state) affect RNA quality.
- To identify appropriate reference gene mRNAs for normalization in schizophrenia brain studies.
Main Methods:
- A matched cohort of 74 subjects (37 schizophrenia/schizoaffective disorder, 37 controls) was analyzed.
- Middle frontal gyrus tissue pH and RNA integrity number (RIN) were assessed.
- Quantitative reverse transcription-polymerase chain reaction measured nine housekeeper genes.
Main Results:
- High RNA integrity numbers (RINs) were observed (mean = 7.3).
- RIN and pH correlated with housekeeper gene expression; agonal state and duration of illness affected some mRNAs.
- Schizophrenia cases showed reduced PPIA and SDHA mRNA, suggesting these are poor normalization candidates.
Conclusions:
- The study cohort was well-matched and adequately powered to detect disease-related mRNA differences.
- Multiple factors, including pH and agonal state, must be considered in human brain mRNA expression studies.
- Well-matched cohorts allow for reliable detection of subtle gene expression differences in schizophrenia.
