Evaluation of CHD5, H3K9me3, and H4K12ac in Human Testes with Spermatogenic Maturation Arrest: A Cross-Sectional Study.

IF 2.3 Q2 OBSTETRICS & GYNECOLOGY International Journal of Fertility & Sterility Pub Date : 2024-06-09 DOI:10.22074/ijfs.2023.1996254.1451
Paisal Paisal, Dwi A Pujianto, Kusmardi Kusmardi, Ponco Birowo, Asmarinah Asmarinah
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Abstract

Background: Spermatogenic maturation arrest is thought to be caused by epigenetic defects, specifically in chromatin remodeling and histone modification. This study evaluated the status of chromatin remodeling chromodomain helicase DNA binding protein 5 (CHD5) and histone modifications histone 4 lys-12 acetylation (H4K12ac) and histone 3 lys-9 trimethylation (H3K9me3) in human testicular biopsies, based on maturation arrest type.

Materials and methods: The cross-sectional study utilized 18 Bouin-fixed paraffin-embedded (BFPE) specimens prepared from residual tissue from routine laboratory tests of infertile patients. The expression of CHD5, H4K12ac, and H3K9me3 was examined through immunohistochemistry (IHC). The intensity was measured using ImageJ with IHC Profiler and StarDist plugins. Statistical analysis was performed using Python with Scipy.Stats module. The data were tested with Shapiro- Wilk for normality and Levene test for homogeneity. The differences in the intensity of spermatogenic cells were assessed using Kruskal-Wallis and Mann-Whitney tests. A difference was considered statistically significant if P<0.05.

Results: We found three types of maturation arrest, including Sertoli cell only (n=5), spermatocyte arrest (n=4), and spermatid arrest (n=9). CHD5 was positive in spermatogonia and round spermatids but absent in spermatocytes. The mean grey value (MGV) of CHD5 in spermatogonia was generally weak in spermatocyte arrest (157.4 ± 16.6) and spermatid arrest (155.3 ± 16.8), and there was no significant difference between them [P=0.49, 95% confidence interval (CI): (-4.3, 6), effect size (r): 0.02]. Although there was a significant difference in the expression of H3K9me3 and H4K12ac (P<0.001), both histone modifications were found in all observed spermatogenic cells.

Conclusion: The expressions of CHD5, H3K9me3, and H4K12ac in different spermatogenic cell types produce similar results, indicating that they cannot be used as markers to determine the type of spermatogenic maturation arrest in humans. The significant finding in this research is the expression of CHD5 in human spermatogonia cells, which requires further study for elaboration.

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人类精子发生成熟停滞睾丸中 CHD5、H3K9me3 和 H4K12ac 的评估:一项横断面研究
背景:生精成熟停滞被认为是由表观遗传缺陷引起的,特别是染色质重塑和组蛋白修饰。本研究根据成熟停滞类型,评估了人类睾丸活检组织中染色质重塑染色质链螺旋酶DNA结合蛋白5(CHD5)和组蛋白修饰组蛋白4 lys-12乙酰化(H4K12ac)和组蛋白3 lys-9三甲基化(H3K9me3)的状况:这项横断面研究使用了 18 份布氏固定石蜡包埋(BFPE)标本,这些标本取自不育患者常规实验室检查的残留组织。通过免疫组织化学(IHC)检测了CHD5、H4K12ac和H3K9me3的表达。使用带有 IHC Profiler 和 StarDist 插件的 ImageJ 对强度进行测量。统计分析使用 Python 的 Scipy.Stats 模块进行。用 Shapiro- Wilk 检验数据的正态性,用 Levene 检验数据的同质性。精原细胞强度的差异采用 Kruskal-Wallis 和 Mann-Whitney 检验进行评估。如果结果有统计学意义,则认为差异显著:我们发现了三种类型的成熟停滞,包括仅Sertoli细胞停滞(5个)、精母细胞停滞(4个)和精子细胞停滞(9个)。CHD5在精原细胞和圆形精母细胞中呈阳性,但在精母细胞中不存在。精原细胞中CHD5的平均灰度值(MGV)在精母细胞停育(157.4 ± 16.6)和精子细胞停育(155.3 ± 16.8)时普遍较弱,两者之间无显著差异[P=0.49,95% 置信区间(CI):(-4.3,6),效应大小(r):0.02]。虽然 H3K9me3 和 H4K12ac 的表达存在显著差异(PConclusion:CHD5、H3K9me3和H4K12ac在不同生精细胞类型中的表达产生了相似的结果,表明它们不能作为判断人类生精成熟停滞类型的标志物。本研究的重要发现是 CHD5 在人类精原细胞中的表达,这还需要进一步研究阐述。
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来源期刊
CiteScore
4.20
自引率
0.00%
发文量
68
审稿时长
>12 weeks
期刊介绍: International Journal of Fertility & Sterility is a quarterly English publication of Royan Institute . The aim of the journal is to disseminate information through publishing the most recent scientific research studies on Fertility and Sterility and other related topics. Int J Fertil Steril has been certified by Ministry of Culture and Islamic Guidance in 2007 and was accredited as a scientific and research journal by HBI (Health and Biomedical Information) Journal Accreditation Commission in 2008. Int J Fertil Steril is an Open Access journal.
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