Novel 3D analysis using optical tissue clearing documents the evolution of murine rapidly progressive glomerulonephritis
Novel 3D analysis using optical tissue clearing documents the evolution of murine rapidly progressive glomerulonephritis
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DOI：
10.1016/j.kint.2019.02.034
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发表时间：
2019-08-01
影响因子：
19.6
通讯作者：
Moeller, Marcus J.
中科院分区：
文献类型：
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作者：
Puelles, Victor G.;Fleck, David;Moeller, Marcus J.
光学组织清除的最新发展已经难以应用于具有高细胞含量和小功能结构的器官（例如肾脏）的形态测量分析。在这里，我们建立了遗传和免疫标记的组合，用于单细胞鉴定，组织清除和随后的组织免疫病理学和透射电子显微镜的澄清。使用先进的光学显微镜和计算分析，我们研究了小鼠模型的新月体肾炎，炎症性肾脏疾病的典型免疫介导的损害肾小球导致形成细胞增生病变和肾毒性血清诱导的肾功能的快速丧失。结果显示肾小球的分级易感性、显著的足细胞损失和毛细血管损伤。这些效应与壁上皮细胞的活化和肾小球病变的形成相关，肾小球病变可能演变并阻塞肾小管，从而解释肾功能丧失。因此，我们的工作为单细胞分辨率的复杂组织分析提供了新的高通量终点。
Recent developments in optical tissue clearing have been difficult to apply for the morphometric analysis of organs with high cellular content and small functional structures, such as the kidney. Here, we establish combinations of genetic and immuno-labelling for single cell identification, tissue clearing and subsequent de-clarification for histoimmunopathology and transmission electron microscopy. Using advanced light microscopy and computational analyses, we investigated a murine model of crescentic nephritis, an inflammatory kidney disease typified by immune-mediated damage to glomeruli leading to the formation of hypercellular lesions and the rapid loss of kidney function induced by nephrotoxic serum. Results show a graded susceptibility of the glomeruli, significant podocyte loss and capillary injury. These effects are associated with activation of parietal epithelial cells and formation of glomerular lesions that may evolve and obstruct the kidney tubule, thereby explaining the loss of kidney function. Thus, our work provides new high-throughput endpoints for the analysis of complex tissues with single-cell resolution.