Abstract
Histone acetylation plays a role in the regulation of gene transcription. Yet it is not known whether post-mortem brain tissue is suitable for the analysis of histone acetylation. To examine this question, nucleosomes were isolated from frontal cortex of nine subjects which were obtained at short times after death and immediately frozen at −80°C or maintained at room temperature from 3 h up to 50 h after death and then frozen at −80°C to mimic variable post-mortem delay in tissue processing as currently occurs in normal practice. Chromatin immunoprecipitation assays were performed for two lysine residues, H3K9ac and H3K27ac. Four gene loci were amplified by SyBrGreen PCR: Adenosine A2A receptor, UCHL1, α-synuclein and β-globin. Results showed variability in the histone acetylation level along the post-mortem times and an increase in the acetylation level at an unpredictable time from one case to another and from one gene to another within the first 24 h of post-mortem delay. Similar results were found with three rat brains used to exclude the effects of agonal state and to normalize the start-point as real time zero. Therefore, the present observations show that human post-mortem brain is probably not suitable for comparative studies of histone acetylation.
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Acknowledgments
We are grateful to Dr. Akbarian and Dr. Huang for nucleosomes isolation protocol as well as helpful comments for ChIP to Input ratio determination. We thank T. Yohannan for editorial assistance. This study was funded by grants from the Ministerio de Ciencia e Innovación, Instituto de Salud Carlos III (CP08/00095 to MB; PI08/0582 to IF), the European Commission under the Sixth Framework Programme (BrainNet Europe II, LSHM-CT-2004-503039) to IF and the Fundació La Marató de TV3 (092330) to MB. IVM is a recipient of an IDIBELL predoctoral fellowship.
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For every specific PCR product in the human artificial post-mortem delays (Fig. 1), the dissociation curve analysis shows lack of formation of primer dimers and unspecific PCR amplifications (DNA: PCR product; NTC: non-template control) (PDF 121 kb)
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Barrachina, M., Moreno, J., Villar-Menéndez, I. et al. Histone tail acetylation in brain occurs in an unpredictable fashion after death. Cell Tissue Bank 13, 597–606 (2012). https://doi.org/10.1007/s10561-011-9278-9
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DOI: https://doi.org/10.1007/s10561-011-9278-9