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

Brain protein preservation largely depends on the postmortem storage temperature: implications for study of proteins in human neurologic diseases and management of brain banks: a BrainNet Europe Study.

Author(s): Ferrer I, Santpere G, Arzberger T, Bell J, Blanco R, Boluda S, Budka H, Carmona M, Giaccone G, Krebs B, Limido L, Parchi P, Puig B, Strammiello R, Ströbel T, Kretzschmar H

Publication: J Neuropathol Exp Neurol, 2007, Vol. 66, Page 35

PubMed ID: 17204935 PubMed Review Paper? No

Purpose of Paper

The purpose of this study was to determine the degree of influence of storage temperature (room temperature, 1 and 4 degrees C) and duration prior to freezing on protein integrity and expression using paired postmortem brain specimens.

Conclusion of Paper

The authors reported that storage at 1 degrees C for up to 50 hours prior to freezing did not permit protein degradation. However, samples stored at 4 degrees C for 16 hours or more resulted in protein-specific degradation with several proteins identified as stable for up to 50 hours. Protein degradation was extensive in samples stored at room temperature for 23 hours or more. Phosphotau degradation was observed in an Alzheimers diagnosed case after 6 hours at room temperature or 50 hours at 4 degrees C.

Studies

  1. Study Purpose

    The purpose of this study was to evaluate the influence of postmortem storage time and temperature on brain proteins. Sections of one biospecimen per storage temperature (which had been obtained approximately two hours after death) was evaluated at numerous times up to 50 hours postmortem.

    Summary of Findings:

    Protein degradation was not observed in samples stored at 1 degrees C for up to 50 hours prior to freezing or in samples stored at 4 degrees C for no more than 8 hours. However, effects of storage at 4 degrees C for 16 to 50 hours yielded protein-specific results with beta-actin, p38-P, proteasome-20 and -11, displaying no signs of degradation. Samples stored at room temperature displayed degradation after 23 hours of storage in a protein-specific manner, with reduced expression for all proteins examined (with the exception of the proteasomes) after 50 hours. Several additional proteins displaying evidence of stability after storage at 4 degrees C for 50 hours were identified via mass spectrometry: YWHAZ, G3PDH, malate dehydrogenase, adolase.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    Protein 1D/2D gels
    Protein LC-MS or LC-MS/MS
    Protein Western blot
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage temperature 1 degrees C
    4 degrees C
    22 degrees C
    23 degrees C
    Storage Storage duration 2 h
    5 h
    8 h
    23 h
    50 h
  2. Study Purpose

    The purpose of this study was to determine possible protein expression effects in sarkosyl-insoluble fractions of Alzheimer brain specimens (a) stored at 4 degrees C or room temperature for various durations or (b) refrozen after a secondary storage at room temperature for 12 to 48 hours.

    Summary of Findings:

    Phosphotau degradation was observed in samples stored at room temperature in a time-dependent manner, beginning as early as 6 hours. Similar degradation was observed in samples stored at 4 degrees C for 50 hours. Specimens that were subjected to a secondary storage at room temperature and were then refrozen lacked a Western blot signal.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Alzheimer's Disease
    Platform:
    AnalyteTechnology Platform
    Protein Western blot
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Time at room temperature 5 h
    8 h
    12 h
    24 h
    26 h
    48 h
    50 h

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