Effect of post-mortem delay on N-terminal huntingtin protein fragments in human control and Huntington disease brain lysates

PLoS One. 2017 Jun 1;12(6):e0178556. doi: 10.1371/journal.pone.0178556. eCollection 2017.


Huntington disease is associated with elongation of a CAG repeat in the HTT gene that results in a mutant huntingtin protein. Several studies have implicated N-terminal huntingtin protein fragments in Huntington disease pathogenesis. Ideally, these fragments are studied in human brain tissue. However, the use of human brain tissue comes with certain unavoidable variables such as post mortem delay, artefacts from freeze-thaw cycles and subject-to-subject variation. Knowledge on how these variables might affect N-terminal huntingtin protein fragments in post mortem human brain is important for a proper interpretation of study results. The effect of post mortem delay on protein in human brain is known to vary depending on the protein of interest. In the present study, we have assessed the effect of post mortem delay on N-terminal huntingtin protein fragments using western blot. We mimicked post mortem delay in one individual control case and one individual Huntington disease case with low initial post mortem delay. The influence of subject-to-subject variation on N-terminal huntingtin fragments was assessed in human cortex and human striatum using two cohorts of control and Huntington disease subjects. Our results show that effects of post mortem delay on N-terminal huntingtin protein fragments are minor in our individual subjects. Additionally, one freeze-thaw cycle decreases the huntingtin western blot signal intensity in the cortex control subject, but does not introduce additional N-terminal huntingtin fragments. Our results suggest that subject-to-subject variation contributes more to variability in N-terminal huntingtin fragments than post mortem delay.

MeSH terms

  • 3' Untranslated Regions
  • Aged
  • Aged, 80 and over
  • Amino Acid Sequence
  • Brain / metabolism*
  • Brain / pathology
  • Case-Control Studies
  • Cell Line
  • Female
  • HEK293 Cells
  • Humans
  • Huntingtin Protein / metabolism*
  • Huntington Disease / metabolism
  • Huntington Disease / pathology*
  • Male
  • Middle Aged
  • Open Reading Frames
  • Postmortem Changes
  • Sequence Homology, Amino Acid


  • 3' Untranslated Regions
  • Huntingtin Protein

Grant support

This study was funded by grant WAR08-08 from the Princess Beatrix Spierfonds (https://www.prinsesbeatrixspierfonds.nl) received by WMCRM. MHS received a travel and expenses grant by the Bontius stichting (https://www.lumc.nl/org/bontius-stichting) to perform part of this work in New Zealand. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.