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小鼠睾丸间质中的端细胞:G蛋白偶联雌激素受体(GPER)和雌激素相关受体(ERR)在小鼠睾丸间质细胞调控中的意义

Telocytes in the mouse testicular interstitium: implications of G-protein-coupled estrogen receptor (GPER) and estrogen-related receptor (ERR) in the regulation of mouse testicular interstitial cells.

作者信息

Pawlicki Piotr, Hejmej Anna, Milon Agnieszka, Lustofin Krzysztof, Płachno Bartosz J, Tworzydlo Waclaw, Gorowska-Wojtowicz Ewelina, Pawlicka Bernadetta, Kotula-Balak Malgorzata, Bilinska Barbara

机构信息

Department of Endocrinology, Institute of Zoology and Biomedical Research, Jagiellonian University in Kraków, Gronostajowa 9, 30-387, Krakow, Poland.

Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University in Kraków, Gronostajowa 9, 30-387, Krakow, Poland.

出版信息

Protoplasma. 2019 Mar;256(2):393-408. doi: 10.1007/s00709-018-1305-2. Epub 2018 Sep 5.

Abstract

Telocytes (TCs), a novel type of interstitial cells, are involved in tissue homeostasis maintenance. This study aimed to investigate TC presence in the interstitium of mouse testis. Additionally, inactivation of the G-coupled membrane estrogen receptor (GPER) in the testis was performed to obtain insight into TC function, regulation, and interaction with other interstitial cells. Mice were injected with a GPER antagonist (G-15; 50 μg/kg bw), and the GPER-signaling effect on TC distribution, ultrastructure, and function, as well as the interstitial tissue interaction of GPER with estrogen-related receptors (ERRs), was examined. Microscopic observations of TC morphology were performed with the use of scanning and transmission electron microscopes. Telocyte functional markers (CD34; c-kit; platelet-derived growth factor receptors α and β, PDGFRα and β; vascular endothelial growth factor, VEGF; and vimentin) were analyzed by immunohistochemistry/immunofluorescence and Western blot. mRNA expression of CD34 as well as ERR α, β, and γ was measured by qRT-PCR. Relaxin and Ca concentrations were analyzed by immunoenzymatic and colorimetric assays, respectively. For the first time, we reveal the presence of TCs in the interstitium together with the peritubular area of mouse testis. Telocytes were characterized by specific features such as a small cell body and extremely long prolongations, constituting a three-dimensional network mainly around the interstitial cells. Expression of all TC protein markers was confirmed. Based on scanning electron microscopic observation in GPER-blocked testis, groups of TCs were frequently seen. No changes were found in TC ultrastructure in GPER-blocked testis when compared to the control. However, tendency to TC number change (increase) after the blockage was observed. Concomitantly, no changes in mRNA CD34 expression and increase in ERR expression were detected in GPER-blocked testes. In addition, Ca was unchanged; however, an increase in relaxin concentration was observed. Telocytes are an important component of the mouse testicular interstitium, possibly taking part in maintaining its microenvironment as well as contractile and secretory functions (via themselves or via controlling of other interstitial cells). These cells should be considered a unique and useful target cell type for the prevention and treatment of testicular interstitial tissue disorders based on estrogen-signaling disturbances.

摘要

端细胞(TCs)是一种新型的间质细胞,参与组织内环境稳态的维持。本研究旨在调查小鼠睾丸间质中端细胞的存在情况。此外,通过使睾丸中的G蛋白偶联膜雌激素受体(GPER)失活,以深入了解端细胞的功能、调节及其与其他间质细胞的相互作用。给小鼠注射GPER拮抗剂(G-15;50μg/kg体重),并检测GPER信号对端细胞分布、超微结构和功能的影响,以及GPER与雌激素相关受体(ERRs)在间质组织中的相互作用。使用扫描电子显微镜和透射电子显微镜对端细胞形态进行微观观察。通过免疫组织化学/免疫荧光和蛋白质印迹分析端细胞功能标志物(CD34、c-kit、血小板衍生生长因子受体α和β、PDGFRα和β、血管内皮生长因子、VEGF和波形蛋白)。通过qRT-PCR检测CD34以及ERRα、β和γ的mRNA表达。分别通过免疫酶法和比色法分析松弛素和钙的浓度。我们首次揭示了小鼠睾丸间质以及睾丸周围区域中端细胞的存在。端细胞具有一些特定特征,如小细胞体和极长的突起,主要围绕间质细胞构成三维网络。所有端细胞蛋白标志物的表达均得到证实。基于对GPER阻断的睾丸的扫描电子显微镜观察,经常可见成群的端细胞。与对照组相比,GPER阻断的睾丸中端细胞超微结构未发现变化。然而,观察到阻断后端细胞数量有增加的趋势。同时,在GPER阻断的睾丸中未检测到mRNA CD34表达的变化,但ERR表达增加。此外,钙含量未变;然而,观察到松弛素浓度增加。端细胞是小鼠睾丸间质的重要组成部分,可能参与维持其微环境以及收缩和分泌功能(通过自身或通过控制其他间质细胞)。基于雌激素信号紊乱,这些细胞应被视为预防和治疗睾丸间质组织疾病的独特且有用的靶细胞类型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae88/6510843/572f12c7738a/709_2018_1305_Fig1_HTML.jpg

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