Deletion of an evolutionarily conserved TAD boundary impacts spermatogenesis in mice†.

IF 3.1 2区 生物学 Q2 REPRODUCTIVE BIOLOGY
Ana C Lima, Mariam Okhovat, Alexandra M Stendahl, Ran Yang, Jake VanCampen, Kimberly A Nevonen, Jarod Herrera, Weiyu Li, Lana Harshman, Lev M Fedorov, Katinka A Vigh-Conrad, Nadav Ahituv, Donald F Conrad, Lucia Carbone
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Abstract

Spermatogenesis is a complex process that can be disrupted by genetic and epigenetic changes, potentially leading to male infertility. Recent research has rapidly increased the number of protein coding mutations causally linked to impaired spermatogenesis in humans and mice. However, the role of non-coding mutations remains largely unexplored. As a case study to evaluate the effects of non-coding mutations on spermatogenesis, we first identified an evolutionarily conserved topologically associated domain (TAD) boundary near two genes with important roles in mammalian testis function: Dmrtb1 and Lrp8. We then used CRISPR-Cas9 to generate a mouse line where 26 kb of the boundary was removed including a strong and evolutionarily conserved CTCF binding site. ChIP-seq and Hi-C experiments confirmed the removal of the CTCF site and a resulting mild increase in the DNA-DNA interactions across the domain boundary. Mutant mice displayed significant changes in testis gene expression, higher frequency of histological abnormalities, a drop of 47-52% in efficiency of meiosis, a 15-18% reduction in efficiency of spermatogenesis, and consistently, a 12-28% decrease in daily sperm production compared to littermate controls. Despite these quantitative changes in testis function, mutant mice show no significant changes in fertility. This suggests that non-coding deletions affecting testis gene regulation may have smaller effects on fertility compared to coding mutations of the same genes. Our results demonstrate that disruption of a TAD boundary can have a negative impact on sperm production and highlight the importance of considering non-coding mutations in the analysis of patients with male infertility.

一个进化上保守的TAD边界的缺失会影响小鼠的精子发生。
精子发生是一个复杂的过程,可能受到遗传和表观遗传变化的干扰,可能导致男性不育。最近的研究迅速增加了与人类和小鼠精子发生受损有因果关系的蛋白质编码突变的数量。然而,非编码突变的作用在很大程度上仍未被探索。作为评估非编码突变对精子发生影响的案例研究,我们首先在两个在哺乳动物睾丸功能中起重要作用的基因Dmrtb1和Lrp8附近发现了一个进化保守的拓扑相关结构域(TAD)边界。然后,我们使用CRISPR-Cas9生成了一个小鼠系,其中26 kb的边界被删除,包括一个强大的和进化上保守的CTCF结合位点。ChIP-seq和Hi-C实验证实了CTCF位点的去除,并由此导致DNA-DNA相互作用在结构域边界上的轻微增加。突变小鼠表现出睾丸基因表达的显著变化,组织学异常的频率更高,减数分裂效率下降47-52%,精子发生效率下降15-18%,每日精子产量与对照组相比持续下降12-28%。尽管睾丸功能发生了这些定量变化,但突变小鼠的生育能力没有明显变化。这表明,与相同基因的编码突变相比,影响睾丸基因调控的非编码缺失可能对生育能力的影响较小。我们的研究结果表明,TAD边界的破坏会对精子产生负面影响,并强调了在分析男性不育症患者时考虑非编码突变的重要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biology of Reproduction
Biology of Reproduction 生物-生殖生物学
CiteScore
6.30
自引率
5.60%
发文量
214
审稿时长
1 months
期刊介绍: Biology of Reproduction (BOR) is the official journal of the Society for the Study of Reproduction and publishes original research on a broad range of topics in the field of reproductive biology, as well as reviews on topics of current importance or controversy. BOR is consistently one of the most highly cited journals publishing original research in the field of reproductive biology.
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