EHD3 positively regulated by NR5A1 participates in testosterone synthesis via endocytosis.
EHD3 positively regulated by NR5A1 participates in testosterone synthesis via endocytosis.
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NR5A1 正向调节的 EHD3 通过内吞作用参与睾酮合成。
DOI：
10.1016/j.lfs.2021.119570
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发表时间：
2021-05
期刊：
影响因子：
6.1
通讯作者：
Runsheng Li
中科院分区：
文献类型：
--
作者：
Lingling Zhang;Lijun Ding;Yifan Li;Fangxi Zhang;Yanhong Xu;Hongjie Pan;Xiaofeng Wan;Guijun Yan;Fei Yu;Runsheng Li
目的越来越多的证据表明，激素的分泌受内吞作用的调节。Eps15同源结构域含蛋白3(EHD3)是一种内吞转运调节蛋白，但EHD3是否与睾酮分泌有关尚不清楚。本研究旨在探讨EHD3在睾酮合成中的作用。主要方法：用双抗体夹心法测定睾酮浓度。用外切体示踪法和免疫荧光法检测EHD3对细胞内吞作用的影响。染色质免疫沉淀(ChIP)和双荧光素酶报告基因检测证实了EHD3和NR5A1之间的靶向关系。对于体内评估，采用CRISPR/Cas9基因打靶技术建立条件NR5A1基因敲除小鼠模型。关键发现EHD3过表达显著增加睾酮浓度。EHD3基因敲除通过减少内吞作用显著减少了睾酮的合成。EHD3启动子的活性受NR5A1正调控，NR5A1占据EHD3启动子的保守序列“AGGTCA”。此外，莱迪格细胞特异性条件性NR5A1基因敲除的小鼠显示出迟钝的EHD3和clathrin(内吞作用的关键因素)水平，以及血清睾酮浓度与NR5A1f/fMICE相比。意义本研究暗示了睾酮合成的潜在分子机制，以全面了解男性生殖健康。
AimsIncreasing evidence has shown that hormone secretion is regulated by endocytosis. Eps15 homology domain-containing protein 3 (EHD3) is an endocytic-trafficking regulatory protein, but whether EHD3 is associated with testosterone secretion is not clear. This work aims to explore the role of EHD3 in testosterone synthesis.Main methodsTestosterone concentration was determined by ELISA. The effects of EHD3 on endocytosis were assessed by exosomes tracing assay and Immunofluorescence. Targeting relationship between EHD3 and NR5A1 was verified by chromatin immunoprecipitation (ChIP) and dual luciferase reporter gene assay in Leydig cells. For in vivo assessments, conditional NR5A1 knockout mouse model was established with CRISPR/Cas9 gene targeting technology.Key findingsEHD3 overexpression significantly increased the concentration of testosterone. EHD3 knockdown markedly decreased testosterone synthesis by reducing endocytosis. The activity of theEHD3promoter was positively regulated by NR5A1, which occupied the conserved sequence “AGGTCA” in theEHD3promoter. Furthermore, mice with a Leydig cell-specific conditional NR5A1 knockout displayed the blunted levels of EHD3 and clathrin (a key factor for endocytosis), and serum testosterone concentration compared with NR5A1f/fmice.SignificanceThis study suggests a potential molecular mechanism of testosterone synthesis to fully understand male reproductive health.
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DOI：
10.1016/0006-291x(82)91116-0
发表时间：
1982-05
影响因子：
3.1
作者：
H. Koenig;A. Goldstone;C. Lu
通讯作者：
H. Koenig;A. Goldstone;C. Lu
DOI：
10.4183/aeb.2018.155
发表时间：
2018-04
期刊：
Acta endocrinologica
影响因子：
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作者：
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通讯作者：
T. Zhang;T. Zheng;C. Wang;W. Zhang;D. Jia;R. Wang;B. Qiao
DOI：
10.1083/jcb.201608071
发表时间：
2017-01-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kadlecova Z;Spielman SJ;Loerke D;Mohanakrishnan A;Reed DK;Schmid SL
通讯作者：
Schmid SL
影响因子：
21.3
作者：
通讯作者：
--
影响因子：
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作者：
D. A. Rice;Andréa;-R.;Mouw;A. M. Bogerd;Keith;-L.;Parker
通讯作者：
D. A. Rice;Andréa;-R.;Mouw;A. M. Bogerd;Keith;-L.;Parker