TNF‐α/ENO1 signaling facilitates testicular phagocytosis by directly activating Elmo1 gene expression in mouse Sertoli cells
TNF‐α/ENO1 signaling facilitates testicular phagocytosis by directly activating Elmo1 gene expression in mouse Sertoli cells
复制标题
DOI：
10.1111/febs.16326
复制
发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
H. Xiong;Zhen-Rui Chen;Jie Zhao;Wei Li;Shun Zhang
中科院分区：
文献类型：
--
作者：
H. Xiong;Zhen-Rui Chen;Jie Zhao;Wei Li;Shun Zhang
凋亡的生殖细胞（GC）以及从发育中的精子细胞释放的残留体的吞噬清除对于支持细胞（SC）维持睾丸内环境的稳态是至关重要的。然而，控制吞噬作用的分子机制不明确。在这里，我们确定了α-烯醇化酶（ENO 1）的新作用，这是糖酵解过程中的一种关键酶，作为一种分子，通过吞噬和细胞运动1（Elmo 1）基因的反式激活促进睾丸吞噬作用。使用免疫组织化学和免疫荧光双标记，观察到ENO 1仅在SC的细胞核中表达，其表达与SC分化的完成相关。通过将TM 4细胞与不同的药理学抑制剂孵育并建立TM 4 Tnfr 1 −/−细胞，我们证明了ENO 1的SC特异性表达受到凋亡GC的精细旁分泌控制。反过来，持续阻断ENO 1表达的验证小干扰RNA协议导致精子发生的干扰和男性生育能力的损害。此外，使用ChIP，电泳迁移率变化和荧光素酶报告基因测定，我们表明，在存在凋亡GC的情况下，ENO 1结合到Elmo 1启动子的远端区域，并促进Elmo 1基因的反式激活。与此一致，ELMO 1的过表达改善了TM 4细胞中ENO 1缺陷诱导的吞噬功能受损。这些数据揭示了一个新的作用，SC-特异性表达的ENO 1在调节睾丸吞噬作用，确定肿瘤坏死因子-α和ELMO 1作为关键的上游和下游因子介导的ENO 1行动，并有重要的意义，我们的理解旁分泌控制SC功能的相邻GC。
Phagocytic clearance of apoptotic germ cells (GCs), as well as residual bodies released from developing spermatids, is critical for Sertoli cells (SCs) to maintain inner environment homeostasis within the testis. However, the molecular mechanisms controlling the phagocytosis are ill defined. Here, we identify a new role for alpha‐enolase (ENO1), a key enzyme during glycolysis, as a molecule that facilitates testicular phagocytosis via transactivation of the engulfment and cell motility 1 (Elmo1) gene. Using immunohistochemistry and double‐labeling immunofluorescence, ENO1 was observed to be expressed exclusively in the nuclei of SCs and its expression correlated with the completion of SC differentiation. By incubating TM4 cells with different pharmacological inhibitors and establishing TM4Tnfr1−/− cells, we demonstrated that SC‐specific expression of ENO1 was under a delicate paracrine control from apoptotic GCs. In turn, persistent blockade of ENO1 expression by a validated small interfering RNA protocol resulted in the disturbance of spermatogenesis and impairment of male fertility. Furthermore, using ChIP, electrophoretic mobility shift and luciferase reporter assays, we showed that, in the presence of apoptotic GCs, ENO1 binds to the distal region of the Elmo1 promoter and facilitates transactivation of the Elmo1 gene. In agreement, overexpression of ELMO1 ameliorated ENO1 deficiency‐induced impairment of phagocytosis in TM4 cells. These data reveal a novel role for SC‐specific expression of ENO1 in regulating phagocytosis in testis, identify tumor necrosis factor‐α and ELMO1 as critical upstream and downstream factors in mediating ENO1 action, and have important implications for our understanding of paracrine control of SC function by adjacent GCs.