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精子DNA甲基化的样本内异质性。

Intra-sample heterogeneity of sperm DNA methylation.

作者信息

Jenkins Timothy G, Aston Kenneth I, Trost Cooper, Farley Jordan, Hotaling James M, Carrell Douglas T

机构信息

Andrology and IVF Laboratories, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA.

Division of Urology, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA.

出版信息

Mol Hum Reprod. 2015 Apr;21(4):313-9. doi: 10.1093/molehr/gau115. Epub 2014 Dec 26.

Abstract

The study of sperm epigenetics is challenging and limited largely to sperm population estimates rather than individual spermatozoa. While this type of approach is likely sufficient for most somatic cell lines, it is problematic in a tissue where a single cell is disproportionality influential. Furthermore, we know very little about the epigenetic variability between different sperm from the same ejaculate. Thus, it is essential that we better understand the heterogeneity of sperm epigenetic marks within an ejaculate. In this study, we have performed sperm genome-wide DNA methylation analyses on single ejaculates from 20 individuals. Sperm samples were subjected to gradient separation, following which the 90% layer ('high-quality sperm') and the 35% layer ('low-quality sperm') were isolated and analyzed separately using the Illumina 450K methylation array. We did not identify any single CpG that was differentially methylated between the two fractions. In contrast, we did identify 772 significant regional methylation alterations between the two layers. Coefficient of variance analysis also revealed that, in addition to having multiple sites that appear to be differentially methylated, the 35% layer sperm population, as a whole, displayed significantly higher variability in DNA methylation than did the 90% layer. In conclusion, while the two sperm populations analyzed here do not appear to be entirely distinct, those sperm that are generally considered to be of 'poor-quality' display some consistent regions of alteration and, more strikingly, demonstrate more heterogeneity than sperm considered to be of more normal quality.

摘要

精子表观遗传学的研究具有挑战性,目前主要局限于对精子群体的估计,而非单个精子。虽然这种方法对于大多数体细胞系可能足够,但在单个细胞具有不成比例影响力的组织中却存在问题。此外,我们对同一射精样本中不同精子之间的表观遗传变异性了解甚少。因此,我们必须更好地理解射精样本中精子表观遗传标记的异质性。在本研究中,我们对20名个体的单个射精样本进行了全基因组DNA甲基化分析。精子样本经过梯度分离,然后分别分离出90%层(“高质量精子”)和35%层(“低质量精子”),并使用Illumina 450K甲基化芯片进行分析。我们没有发现任何单个CpG在这两个部分之间存在差异甲基化。相反,我们确实在这两层之间鉴定出772个显著的区域甲基化改变。方差系数分析还显示,除了有多个位点似乎存在差异甲基化外,35%层的精子群体作为一个整体,其DNA甲基化的变异性明显高于90%层。总之,虽然这里分析的两个精子群体似乎并非完全不同,但那些通常被认为“质量差”的精子显示出一些一致的改变区域,更引人注目的是,与被认为质量更正常的精子相比,它们表现出更多的异质性。

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