Abstract
There has been a growing interest in the study of epigenetic mechanisms to elucidate the molecular bases of human brain-related diseases and disorders. Frequently, researchers utilize post-mortem tissue with the assumption that post-mortem tissue decay has little or no effect on epigenetic marks. Although previous studies show no effect of post-mortem interval on certain epigenetic marks, no such research has been performed on cytosine modifications. In this study, we use DNA from the brains of adult Sprague Dawley rats subjected to post-mortem intervals at room temperature, ranging from 0 to 96 h, to assess the stability of cytosine modifications, namely 5-methycytosine and 5-hydroxymethylcytosine. Our results indicate that neither global nor site-specific levels of 5-methycytosine and 5-hydroxymethylcytosine are affected by the post-mortem intervals we studied. As such, the use of post-mortem tissue to study cytosine modifications in the context of neurological or neuropsychiatric disorders is appropriate.
Highlights
There has been growing interest in the investigation of epigenetic changes associated with disease states
We provide evidence that these cytosine modifications are stable during post-mortem intervals and can reflect the actual status of 5mC and 5hmC at the time of death in post-mortem tissue
DNA methylation was previously shown to withstand various changes in pH with no differences observed even in extremes (Ernst et al, 2008). Since this covalent bond is maintained during cytosine oxidation, the stability of the demethylation intermediates should remain stable across increasing post-mortem interval (PMI)
Summary
There has been growing interest in the investigation of epigenetic changes associated with disease states. Interesting research provided evidence showing that 5mC can be oxidized to 5-hydroxymethylcytosine (5hmC) in a reaction catalyzed by the ten–eleven translocation (TET) enzymes (Kriaucionis and Heintz, 2009; Tahiliani et al, 2009; Ito et al, 2010). This finding led to the discovery of additional oxidative products, namely 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). We provide evidence that these cytosine modifications are stable during post-mortem intervals and can reflect the actual status of 5mC and 5hmC at the time of death in post-mortem tissue
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