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High-resolution quantification of specific mRNA levels in human brain autopsies and biopsies.

Author(s): Castensson A, Emilsson L, Preece P, Jazin EE

Publication: Genome Res, 2000, Vol. 10, Page 1219

PubMed ID: 10958640 PubMed Review Paper? No

Purpose of Paper

This paper conducted an assessment of mRNA expression of 16 genes in brain to determine the effects of sex, gender, age at death, and postmortem interval (PMI); to compare biospecimens obtained via biopsy and autopsy; and to determine the number of cases required to detect 2- to 10-fold statistically relevant differences in gene expression.

Conclusion of Paper

The authors report that, of the 16 genes evaluated, only CYP26A1 showed a significant difference with respect to postmortem interval; only 5-HT-1E showed a significant difference in males and females; and only CYC1A1 levels correlated with age at death. Although mRNA levels were lower in autopsy than in biopsy material, when adjusted to reference genes, differences in relative expression were not significant. Power calculations revealed that studies of genes with low between-individual variability would require approximately ten subjects per group to accurately detect two-fold differences in expression. For genes with a high degree of between-individual variability, however, accurately detecting two-fold differences in expression would require 50 to 60 subjects per group.

Studies

  1. Study Purpose

    The purpose of this study was to evaluate differences in mRNA levels with respect to postmortem interval (PMI), age at death, and sex.

    Summary of Findings:

    The authors report that, of the 16 genes evaluated CYP26A1 alone was significantly affected by postmortem interval, with decreased expression at a longer PMI; 5-HT-1E alone differed significantly among in males and females, with 40% reduced expression in males compared to females; and CYP1A1 alone was significantly impacted by age at death, with decreased expression in older individuals.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA Spectrophotometry
    RNA Electrophoresis
    RNA Real-time qRT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Preaquisition Postmortem interval 34 +/- 10 h
    73 +/- 17 h
    Preaquisition Patient gender Female
    Male
    Preaquisition Patient age 53 +/- 17 years
    79 +/- 7 years
    View more
  2. Study Purpose

    The purpose of this study was to compare the mRNA levels of 16 genes in biopsy and autopsy specimens.

    Summary of Findings:

    Quantified mRNA levels for all 16 genes were lower in autopsy compared to biopsy specimens; however when normalized to ACTB and GAPD levels differences were not significant, suggesting patterns of expression were similar for autopsy and biopsy specimens.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA Electrophoresis
    RNA Real-time qRT-PCR
    RNA Spectrophotometry
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Method of tissue acquisition Autopsy
    Biopsy
    View more
  3. Study Purpose

    In this study, the authors performed power calculations to determine the number of cases needed to detect 2-fold statistically relevant differences among groups examining the influence of postmortem interval, patient age, and patient sex on quantifiable mRNA levels.

    Summary of Findings:

    Results of power calculations employing the data set used to determine the extent of influence patient and biospecimen variables have on the expression of the 16 genes analyzed (13 cases per group), a statistically significant 2-fold difference could be achieved with as few as 9 (for 5-HT-1F) of as many as 50 (for 5-HT-2A) cases per group. However, for a statistically relevant 5-fold difference in expression, 10 cases per group would be required.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA Real-time qRT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Preaquisition Postmortem interval 34 +/- 10 h
    73 +/- 17 h
    Preaquisition Patient gender Female
    Male
    Preaquisition Patient age 53 +/- 17 years
    79 +/- 7 years
    View more

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