The Nuclear DNA Base 5-Hydroxymethylcytosine Is Present in Purkinje Neurons and the Brain

TLDR

Epigenetic regulation is crucial in neurological disorders, yet neuronal chromatin remains poorly understood, with Purkinje neurons exhibiting large euchromatic nuclei compared to the smaller, heterochromatic granule cells. The authors employed thin‑layer chromatography, high‑pressure liquid chromatography, and mass spectrometry to identify the nucleotide as 5‑hydroxymethyl‑2′‑deoxycytidine (hmdC). hmdC accounts for 0.6 % of nucleotides in Purkinje cells, 0.2 % in granule cells, is absent in cancer cell lines, and its high abundance in brain DNA implies a role in neuronal epigenetic regulation.

Abstract

Despite the importance of epigenetic regulation in neurological disorders, little is known about neuronal chromatin. Cerebellar Purkinje neurons have large and euchromatic nuclei, whereas granule cell nuclei are small and have a more typical heterochromatin distribution. While comparing the abundance of 5-methylcytosine in Purkinje and granule cell nuclei, we detected the presence of an unusual DNA nucleotide. Using thin-layer chromatography, high-pressure liquid chromatography, and mass spectrometry, we identified the nucleotide as 5-hydroxymethyl-2'-deoxycytidine (hmdC). hmdC constitutes 0.6% of total nucleotides in Purkinje cells, 0.2% in granule cells, and is not present in cancer cell lines. hmdC is a constituent of nuclear DNA that is highly abundant in the brain, suggesting a role in epigenetic control of neuronal function.