NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

Human postmortem tissue: what quality markers matter?

Author(s): Stan AD, Ghose S, Gao XM, Roberts RC, Lewis-Amezcua K, Hatanpaa KJ, Tamminga CA

Publication: Brain Res, 2006, Vol. 1123, Page 1

PubMed ID: 17045977 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine suitable RNA and protein quality markers and their relationship to tissue pH, postmortem interval (PMI), and thaw parameters in autopsy and surgically resected brain specimens.

Conclusion of Paper

The authors concluded that RNA integrity number (RIN) was the optimal marker of RNA quality, and that brain pH was a good predictor of peri-mortem tissue quality but was not reflective of degradation induced by thawing and refreezing. Protein concentration, as measured by Western blot, was stable across all parameters investigated. RNA quality, determined by RIN, was similar among postmortem and surgical specimens.

Studies

  1. Study Purpose

    The purpose of this study was to evaluate the relationship, if any, between biospecimen characteristics, such as postmortem interval, diagnosis, thaw parameters, or pH, and the quality of extracted RNA and proteins in autopsy and surgically resected brain specimens.

    Summary of Findings:

    RIN was positively correlated to brain pH at low RIN values although no relationship to postmortem interval (up to 40 h) was observed. pH did not differ among regions located at different brain depths, nor was it dependent upon diagnosis. Further, no relationship between PMI and brain pH was observed. A slow thaw and storage (at room temperature for 24 h) of previously frozen brain specimens adversely affected RIN but not brain pH. Assessment in surgically resected brain specimens frozen within 60 minutes of resection yielded variable pH and RIN values, with some cases displaying values equivalent or better than postmortem samples. Protein concentration did not differ due to PMI, pH, or RIN, as measured in a diverse representative protein set (Rab 3A, Syntaxin, Synaptotagmin, Munc).

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Schizophrenia
    • Bipolar Disorder
    • Autopsy
    • Epilepsy
    • Normal
    • Depression
    Platform:
    AnalyteTechnology Platform
    RNA Automated electrophoresis/Bioanalyzer
    Protein Western blot
    RNA Electrophoresis
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Biospecimen location Dorsolateral prefrontal cortex
    Medial prefrontal cortex
    Deep prefrontal cortex
    Biospecimen Aliquots and Components pH 6.7 +/- 0.4
    Preaquisition Postmortem interval 6 - 40 h
    Biospecimen Acquisition Method of tissue acquisition Autopsy
    Surgical resection

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