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24 July 2014 Comparison of the Absolute Level of Epigenetic Marks 5-Methylcytosine, 5-Hydroxymethylcytosine, and 5-Hydroxymethyluracil Between Human Leukocytes and Sperm
Jolanta Guz, Daniel Gackowski, Marek Foksinski, Rafal Rozalski, Ryszard Olinski
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Abstract

5-Methylcytosine is one of the most important epigenetic modifications and has a profound impact on embryonic development. After gamete fusion, there is a widespread and rapid active demethylation process of sperm DNA, which suggests that the paternal epigenome has an important role during embryonic development. To better understand the epigenome of sperm DNA and its possible involvement in a developing embryo, we determined epigenetic marks in human sperm DNA and in surrogate somatic tissue leukocytes; the analyzed epigenetic modifications included 5-methyl-2′-deoxycytidine, 5-hydroxymethyl-2′-deoxycytidine, and 5-hydroxymethyl-2′-deoxyuridine. For absolute determination of the modification, we used liquid chromatography with UV detection and tandem mass spectrometry techniques with isotopically labeled internal standards. Our analyses demonstrated, for the first time to date, that absolute global values of 5-methyl-2′-deoxycytidine, 5-hydroxymethyl-2′-deoxycytidine, and 5-hydroxymethyl-2′-deoxyuridine in sperm are highly statistically different from those observed for leukocyte DNA, with respective mean values of 3.815% versus 4.307%, 0.797 versus 2.945 per 104 deoxynucleosides, and 5.209 versus 0.492 per 106 deoxynucleosides. We hypothesize that an exceptionally high value of 5-hydroxymethyluracil in sperm (>10-fold higher than in leukocytes) may play a not yet recognized regulatory role in the paternal genome.

Jolanta Guz, Daniel Gackowski, Marek Foksinski, Rafal Rozalski, and Ryszard Olinski "Comparison of the Absolute Level of Epigenetic Marks 5-Methylcytosine, 5-Hydroxymethylcytosine, and 5-Hydroxymethyluracil Between Human Leukocytes and Sperm," Biology of Reproduction 91(3), (24 July 2014). https://doi.org/10.1095/biolreprod.114.121541
Received: 16 May 2014; Accepted: 1 July 2014; Published: 24 July 2014
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