Integrative Proteomic and Phosphoproteomic Profiling of Testis from Wip1 Phosphatase-Knockout Mice: Insights into Mechanisms of Reduced Fertility
Integrative Proteomic and Phosphoproteomic Profiling of Testis from Wip1 Phosphatase-Knockout Mice: Insights into Mechanisms of Reduced Fertility
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Wip1 磷酸酶敲除小鼠睾丸的综合蛋白质组学和磷酸化蛋白质组学分析：洞察生育力降低的机制
DOI：
10.1074/mcp.ra117.000479
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发表时间：
2019-02-01
影响因子：
7
通讯作者：
Li, Kui
中科院分区：
文献类型：
--
作者：
Wei, Yinghui;Gao, Qian;Li, Kui
缺乏野生型p53诱导的磷酸酶1（Wip 1）的小鼠表现出雄性生殖缺陷，包括较小的睾丸，生育力低下和圆形和伸长精子细胞阶段的精子发生缺陷。然而，这些异常的分子机制仍不清楚。在这里，我们研究了蛋白质组和磷酸化蛋白质组的Wip 1基因敲除小鼠睾丸使用定量蛋白质组学的方法。从总共鉴定的6872个蛋白质和4280个磷酸化位点中，发现58个蛋白质和159个磷酸化位点与野生型小鼠相比受到差异调节。途径富集分析显示，这些受调节的蛋白和磷酸化位点主要参与粘附/紧密连接、凋亡、炎症反应、精子发生、精子运动以及细胞骨架组装和解聚。Wip 1基因敲除小鼠表现出连接相关蛋白（闭合蛋白、ZO-1和N-钙粘蛋白）的表达减少，血睾屏障的完整性受损。此外，Wip 1缺陷与睾丸中细胞因子和生殖细胞凋亡水平升高相关。这些结果表明，促炎细胞因子可能通过降低连接相关蛋白的表达而损害血-睾丸屏障动力学，这可能导致生育力低下和精子发生缺陷。总的来说，这些发现有助于解释Wip 1缺失导致的生殖功能低下，并为我们理解男性不育的原因提供了新的见解。
Mice lacking wild-type p53-induced phosphatase 1 (Wip1) display male reproductive defects including smaller testes, subfertility and spermatogenesis defects at the round-and elongating-spermatid stages. However, the molecular mechanisms underlying these abnormalities remain unclear. Here we examined the proteome and phosphoproteome of testes from Wip1-knockout mice using a quantitative proteomic approach. From a total of 6872 proteins and 4280 phosphorylation sites identified, 58 proteins and 159 phosphorylation sites were found to be differentially regulated compared with wild type mice. Pathway enrichment analyses revealed that these regulated proteins and phosphosites were mainly involved in adherens/tight junctions, apoptosis, inflammatory response, spermatogenesis, sperm motility, and cytoskeletal assembly and depolymerization. Wip1-knockout mice showed decreased expression of junction-associated proteins (occludin, ZO-1, and N-cadherin) and impaired integrity of the blood-testis barrier. In addition, Wip1 deficiency was associated with elevated levels of cytokines and germ cell apoptosis in the testis. These results suggest that proinflammatory cytokines may impair the blood-testis barrier dynamics by decreasing the expression of junction-associated proteins, which could lead to subfertility and spermatogenesis defects. Collectively, these findings help to explain the low reproductive function caused by Wip1 deletion and provide novel insights into our understanding of causes of male infertility.