Re-characterization of the Glomerulopathy in CD2AP Deficient Mice by High-Resolution Helium Ion Scanning Microscopy.
Re-characterization of the Glomerulopathy in CD2AP Deficient Mice by High-Resolution Helium Ion Scanning Microscopy.
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DOI：
10.1038/s41598-017-08304-3
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发表时间：
2017-08-16
影响因子：
4.6
通讯作者：
Lu HAJ
中科院分区：
文献类型：
--
作者：
Tsuji K;Păunescu TG;Suleiman H;Xie D;Mamuya FA;Miner JH;Lu HAJ
氦离子扫描显微镜（HIM）是一种无需表面涂层即可直接观察细胞表面超微结构的新技术。尽管其分辨率非常高，但尚未广泛应用于研究生物或病理学样品。在这里，我们报告了应用这种强大的技术来检查的三维超微结构特征的蛋白尿性肾小球疾病的小鼠与CD 2相关蛋白（CD 2AP）的缺陷。HIM显示肾小球特征的一系列改变，包括裂隙隔膜的消失和解体，随后是足突消失，主要突起变平和融合，并最终转化为足细胞片。过滤狭缝孔的数量和尺寸减少。引人注目的是，观察到许多“水泡”形的微突起从足细胞突起和细胞体延伸，表明伴随CD 2AP缺乏的显著的膜动力学。可视化肾小球内皮和足细胞-内皮界面揭示了内皮损伤的存在，并破坏了6周龄Cd 2ap-KO小鼠的足细胞和内皮完整性。我们使用HIM技术以纳米级分辨率研究缺乏关键性狭缝血管相关蛋白CD 2AP的小鼠肾小球滤过装置的超微结构改变，突出了HIM提供肾小球疾病生物学和（病理）生理学新见解的巨大潜力。
Helium ion scanning microscopy (HIM) is a novel technology that directly visualizes the cell surface ultrastructure without surface coating. Despite its very high resolution, it has not been applied extensively to study biological or pathology samples. Here we report the application of this powerful technology to examine the three-dimensional ultrastructural characteristics of proteinuric glomerulopathy in mice with CD2-associated protein (CD2AP) deficiency. HIM revealed the serial alteration of glomerular features including effacement and disorganization of the slit diaphragm, followed by foot process disappearance, flattening and fusion of major processes, and eventual transformation into a podocyte sheet as the disease progressed. The number and size of the filtration slit pores decreased. Strikingly, numerous “bleb” shaped microprojections were observed extending from podocyte processes and cell body, indicating significant membrane dynamics accompanying CD2AP deficiency. Visualizing the glomerular endothelium and podocyte-endothelium interface revealed the presence of endothelial damage, and disrupted podocyte and endothelial integrity in 6 week-old Cd2ap-KO mice. We used the HIM technology to investigate at nanometer scale resolution the ultrastructural alterations of the glomerular filtration apparatus in mice lacking the critical slit diaphragm-associated protein CD2AP, highlighting the great potential of HIM to provide new insights into the biology and (patho)physiology of glomerular diseases.
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通过块面扫描电子显微镜揭示的足细胞的三维结构。
影响因子：
4.6
作者：
Ichimura K;Miyazaki N;Sadayama S;Murata K;Koike M;Nakamura K;Ohta K;Sakai T
通讯作者：
Sakai T
影响因子：
18.2
作者：
Greka A;Mundel P
通讯作者：
Mundel P
影响因子：
13.6
作者：
Dessapt-Baradez, Cecile;Woolf, Adrian S.;Gnudi, Luigi
通讯作者：
Gnudi, Luigi
影响因子：
3
作者：
Dávila-Esqueda, ME;Vertiz-Hernández, AA;Martínez-Morales, F
通讯作者：
Martínez-Morales, F
影响因子：
4.6
作者：
Joens, Matthew S.;Chuong Huynh;Kasuboski, James M.;Ferranti, David;Sigal, Yury J.;Zeitvogel, Fabian;Obst, Martin;Burkhardt, Claus J.;Curran, Kevin P.;Chalasani, Sreekanth H.;Stern, Lewis A.;Goetze, Bernhard;Fitzpatrick, James A. J.
通讯作者：
Fitzpatrick, James A. J.