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.

在本提案中，我们将进一步调查先前资助造成的鱼精蛋白 1 和鱼精蛋白 2 缺乏的男性。在我们的初步实验中，我们已经证明精子表现出 DNA 损伤，并且过渡蛋白和组蛋白保留在 DNA 上。然而，与缺乏鱼精蛋白1或鱼精蛋白2的动物相比，精子仍然能够使卵母细胞受精。有趣的是，胚胎停滞在 4 和 8 细胞阶段之间。我们假设所描述的早期基因再激活波因染色质结构的改变（更多的组蛋白、TNP 和 DNA）而改变。 我们将使用 ChIP-seq 来研究精子 DNA 中组蛋白保留是否发生了特异性改变。这表明精子发生过程中组蛋白与鱼精蛋白的交换存在层次结构。众所周知，在野生型动物中，在精子 DNA 中发现了“发育重要基因”的组蛋白，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。我们建议研究 4 和 8 细胞阶段的早期基因再激活。通过比较野生型和 doubleHET 动物，我们想要分析这种基因重新激活如何变化。通过与 ChIP-Seq 数据集关联，我们能够将受损的基因表达与异常的组蛋白保留关联起来。这些结果可以为因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。