Yonsei Med J.  2014 Sep;55(5):1333-1340. 10.3349/ymj.2014.55.5.1333.

Histone Acetylation Level and Histone Acetyltransferase/Deacetylase Activity in Ejaculated Sperm from Normozoospermic Men

• Kim JH
• Jee BC
• Lee JM
• Suh CS
• Kim SH

• KMID: 1799499
• DOI: http://doi.org/10.3349/ymj.2014.55.5.1333

Abstract

PURPOSE
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.
MATERIALS AND METHODS
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.
RESULTS
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).
CONCLUSION
Our works demonstrated a significant relationship of acetylation-associated enzyme activity and strict morphology or sperm DFI.

Keyword

Acetylation; acetyltransferase; deacetylase; histone; sperm

MeSH Terms

Acetylation
Adult
DNA Fragmentation
Epigenesis, Genetic
Histone Acetyltransferases/*metabolism
Histones/*metabolism
Humans
Immunohistochemistry
Male
Middle Aged
Protein Processing, Post-Translational
Semen Analysis
Spermatozoa/*metabolism
Histone Acetyltransferases
Histones

Figure

• 

  Fig. 1 Representative microphotographs showing ejaculated sperm immnunostained by fluorescent antibody for histone H3K9, H3K14, H4K5, H4K8, H4K12, and H4K16 acetylation (×1000). Thick arrows indicate 'strong' intensity and thin arrows indicate 'weak' intensity; All sperm are stained with either 'weak' or 'strong' signal intensity, and immunopositivity was determined as the percentage of sperm showing 'strong' signal per total counted sperms. Negative controls were performed in the absence of primary antibody. Counterstaining was performed using a mounting medium with DAPI. FITC, fluorescein isothiocyanate.

• 

  Fig. 2 Detection of histone acetyltransferase (HAT) and histone deacetylase 1 (HDAC1) in mature human sperm. Western blot assay demonstrates 50 kDa band of HAT and 62 kDa band of HDAC1 in mature human sperm lysates obtained from five men.

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