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Limited predictability of postmortem human brain tissue quality by RNA integrity numbers
Kai-C Sonntag1, George Tejada1, Sivan Subburaju1
1Division of Basic Neuroscience, Department of Psychiatry, Harvard Brain Tissue Resource Center, McLean Hospital/Harvard Medical School, Belmont, Massachusetts, USA.
Journal of Neurochemistry
|April 11, 2016
Summary
RNA integrity numbers (RINs) do not fully predict mRNA integrity in postmortem brain tissue. While RINs indicate mRNA quantity, they do not reliably assess degradation, suggesting a need for more comprehensive quality control methods.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- The RNA integrity number (RIN) is a standard metric for assessing RNA quality, particularly in postmortem human brain samples.
- Concerns exist that RIN values may not accurately reflect messenger RNA (mRNA) integrity or degradation levels.
Purpose of the Study:
- To investigate the correlation between RIN values and mRNA integrity in postmortem human brain tissue.
- To determine if RIN is a reliable indicator of mRNA degradation and quality.
Main Methods:
- Utilized Agilent bioanalyzer and quantitative reverse transcription PCR (qRT-PCR) to analyze RNA.
- Evaluated the expression of 3'-5' mRNA sequences of the cytochrome C-1 (CYC1) gene.
- Assessed RNA and cDNA profiles, and analyzed qRT-PCR cycle thresholds.
Main Results:
- RINs showed inconsistent correlations with RNA and cDNA profiles, indicating poor prediction of cDNA quality.
- Positive correlations were observed between RINs and the amplification of full-length transcripts.
- RINs indicated mRNA quantity independent of degradation but did not predict mRNA integrity.
Conclusions:
- RIN is an incomplete measure of postmortem human brain tissue quality.
- RIN values do not reliably predict mRNA integrity or degradation.
- Relying solely on RIN for quality assessment of postmortem brain samples may be misleading.