Protamine1/Protamine-2 double deficient male mice. Detailed analysis of infertility using ChiP-seq on histones of sperm and scRNA-seq. on early embryos to detail the epigenetic disturbances leading to aberrant gene -reactivation.

鱼精蛋白 1/鱼精蛋白 2 双缺陷雄性小鼠。

• 批准号: 415330713
• 负责人: Professor Dr. Hubert Schorle
• 金额: --
• 依托单位: Institut für Pathologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/415330713?language=en
• 关键词: Protamine1; Protamine; double; deficient; male

In this proposal, we will further investigate protamine1 and protamine2 deficient males resulting from previous grants. In our preliminary experiments, we have shown that the sperm exhibit DNA damage and transition proteins and histones remain on the DNA. However, compared to the Protamine1 or Protamine2 deficient animals, the sperm are still able to fertilize oocytes. Interestingly, the embryos arrest between the 4 and 8 cell stages. We hypothesize that the described early gene reactivation waves are altered by the altered chromatin structure (more histones and TNP's and the DNA). We will use ChIP-seq to investigate whether histone retention is specifically altered in sperm DNA. This would suggest a hierarchy in histone to protamine exchange during spermiogenesis. It is known that in wild-type animals, histones at "developmentally-important genes" are found at the sperm DNA and it is speculated that these histones are removed first after fertilization to ensure rapid reactivation of these genes. We propose to study early gene reactivation at 4 and 8 cell stages. By comparing wild type with doubleHET animals we want to analyze how this gene reactivation changes. By correlating with the ChIP-Seq dataset, we are able to correlate the impaired gene expression with aberrant histone retention. The results could provide valuable insights for patients with fertility disorders due to shifts in protamine ratio or protamine deficiency.

在这项建议中，我们将进一步调查鱼精蛋白1和鱼精蛋白2缺乏男性造成以前的赠款。在我们的初步实验中，我们已经表明，精子表现出DNA损伤和过渡蛋白和组蛋白保留在DNA上。然而，与Protamine 1或Protamine 2缺陷动物相比，精子仍然能够使卵母细胞受精。有趣的是，胚胎在4至8个细胞阶段之间停滞。我们假设所描述的早期基因再激活波是由改变的染色质结构（更多的组蛋白和TNP和DNA）改变的。 我们将使用ChIP-seq来研究组蛋白保留是否在精子DNA中发生了特异性改变。这表明在精子发生过程中组蛋白与鱼精蛋白的交换是有层次的。已知在野生型动物中，在精子DNA中发现了“发育重要基因”处的组蛋白，并且推测这些组蛋白在受精后首先被去除，以确保这些基因的快速重新激活。我们建议在4和8细胞阶段研究早期基因再激活。通过比较野生型和doubleHET动物，我们想分析这种基因再激活的变化。通过与ChIP-Seq数据集相关联，我们能够将受损的基因表达与异常的组蛋白保留相关联。该结果可以为由于鱼精蛋白比例变化或鱼精蛋白缺乏而导致生育障碍的患者提供有价值的见解。