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  • Histone acetylation level and histone acetyltransferase/deacetylase activity in ejaculated sperm from normozoospermic men.

Histone acetylation level and histone acetyltransferase/deacetylase activity in ejaculated sperm from normozoospermic men.

Yonsei medical journal (2014-07-23)
Jee Hyun Kim, Byung Chul Jee, Jang Mi Lee, Chang Suk Suh, Seok Hyun Kim
초록

The aim of this work was to evaluate nuclear histone acetylation level and total histone acetyltransferase (HAT) and deacetylase (HDAC) activity in ejaculated sperm and their relevance to conventional sperm parameters. Thirty-three normozoospermic men were included in this study. Semen samples were processed by swim-up and then immunostained by six acetylation antibodies (H3K9ac, H3K14ac, H4K5ac, H4K8ac, H4K12ac, and H4K16ac). Our preliminary study verified the expression of HAT/HDAC1 in mature human sperm. From vitrified-warmed sperm samples, total HAT/HDAC activity was measured by commercially available kits. Nuclear DNA integrity was also measured by TUNEL assay. The levels of six acetylation marks were not related with conventional sperm parameters including sperm DNA fragmentation index (DFI) as well as HAT/HDAC activity. However, sperm DFI was positively correlated with HAT activity (r=0.038 after adjustment, p<0.02). HAT activity showed a negative relationship with HDAC activity (r=-0.51, p<0.01). Strict morphology was negatively correlated with acetylation enzyme index (=HAT activity/HDAC activity) (r=-0.53, p<0.01). Our works demonstrated a significant relationship of acetylation-associated enzyme activity and strict morphology or sperm DFI.

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Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Citrate Concentrated Solution, BioUltra, for molecular biology, 1 M in H2O
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Citrate Concentrated Solution, BioReagent, suitable for coagulation assays, 4 % (w/v)
Supelco
Sodium Citrate, Pharmaceutical Secondary Standard; Certified Reference Material