Investigation of testis proteome alterations associated with male infertility in <i>Dcaf17</i>-deficient mice

Disruption of <i>Dcaf17</i> in mice resulted in male infertility with severe spermatogenesis defects. To investigate the molecular basis of infertility phenotype, we examined testicular proteomes of wild-type (WT) and <i>Dcaf17^-/-</i> mice using a mass spectrometry-based approach. We identified 727 and 525 differentially expressed proteins (DEPs) in 3- and 8-week old testes of <i>Dcaf17^-/-</i> mice, respectively, with an adjusted p-value cut-off of ≤ 0.05. Among these, 299 and 298 DEPs had fold change of ≥ 1.5 between WT and <i>Dcaf17^-/-</i> testes at ­3- and 8-week old, respectively. In the 3-week old <i>Dcaf17^-/-</i> testes, 59.5% of the DEPs were up-regulated, while 40.5% were down-regulated. Similarly, in the 8-week old <i>Dcaf17^-/-</i> testes, 83.9% and 16.1% DEPs were up-regulated and down-regulated, respectively. Functional annotation and network analyses highlighted that many DEPs were associated with key biological processes, including ubiquitination, RNA processing, translation, protein folding, protein stabilization, metabolic processes, oxidation-reduction processes and sper­matogenesis. Subsequent immunohistochemistry and immunoblotting analyses showed higher ubiquitin levels in <i>Dcaf17^-/-</i> testes compared to WT, suggesting potential impairment in ubiquitin proteasome system (UPS) due to DCAF17 loss of function. Our data provide a basis for further work to elucidate the molecular function(s) of DCAF17 in spermatogenesis and male fertility.

在小鼠中敲除Dcaf17基因（Dcaf17）可导致雄性不育，并伴随严重的精子发生缺陷。为探究该不育表型的分子机制，我们采用基于质谱的分析方法，检测了野生型（Wild-type, WT）与Dcaf17^-/-小鼠的睾丸蛋白质组。我们以校正后P值≤0.05作为筛选阈值，分别在3周龄和8周龄的Dcaf17^-/-小鼠睾丸中鉴定出727和525种差异表达蛋白（differentially expressed proteins, DEPs）。其中，在3周龄和8周龄样本中，分别有299和298种DEPs在野生型与Dcaf17^-/-小鼠睾丸间的表达倍数变化≥1.5。在3周龄的Dcaf17^-/-小鼠睾丸中，59.5%的DEPs呈上调表达，40.5%呈下调表达。类似地，在8周龄的Dcaf17^-/-小鼠睾丸中，83.9%的DEPs为上调表达，16.1%为下调表达。功能注释与网络分析结果显示，多数DEPs与关键生物学过程密切相关，包括泛素化、RNA加工、蛋白质翻译、蛋白质折叠、蛋白质稳定、代谢过程、氧化还原过程以及精子发生。后续免疫组织化学与免疫印迹分析结果显示，Dcaf17^-/-小鼠睾丸中的泛素水平高于野生型小鼠，提示DCAF17功能缺失可能导致泛素-蛋白酶体系统（ubiquitin proteasome system, UPS）功能受损。本研究数据为后续阐明DCAF17在精子发生与雄性生育过程中的分子功能提供了研究基础。

搜集汇总

背景与挑战

背景概述

该数据集通过质谱分析研究了Dcaf17缺陷小鼠睾丸蛋白质组的变化，以探究雄性不育的分子机制。在3周和8周龄小鼠中分别识别出数百个差异表达蛋白，其中许多与泛素化、代谢过程和精子发生等关键生物过程相关，并发现泛素蛋白酶体系统可能因DCAF17功能丧失而受损。这些结果为理解DCAF17在精子发生和雄性生育中的作用提供了重要基础。