Ali Adilijiang, Liu Zixuan, Ye Kenan, Guan Yun, Chen Siying, Liu Tingxuan, Guo Ziyu, Wong Madeline K, Vasquez Pedro, Poudel Chetan, Mustonen Benjamin C, Eng Diana G, Pippin Jeffrey W, Shankland Stuart J, Wang Sheng, Vaughan Joshua C
Department of Chemistry, University of Washington, Seattle, Washington, USA.
Paul G. Allen School of Computer Science & Engineering, University of Washington, Seattle, Washington, USA.
bioRxiv. 2024 Dec 9:2024.10.31.620364. doi: 10.1101/2024.10.31.620364.
Renal glomeruli have traditionally been studied by micrometer-scale optical microscopy to interrogate overall physiology or molecular distributions and by nanoscale electron microscopy to interrogate the ultrastructure of thin sections. While these approaches are powerful, they have been limited in their ability to obtain detailed views of the glomeruli as holistic 3D functional units. To fill this knowledge gap, we have developed a novel pipeline for imaging, reconstructing, and analyzing whole mouse glomeruli at 100 nm resolution using super-resolution fluorescence microscopy. This pipeline integrates both manual and machine learning approaches to annotate and analyze glomerular structures. Using this method, we created 18 detailed glomerulus models, from a range of healthy, aged, and model diseased mice, that outline all major structures and cell types. These models have been made publicly accessible in an online repository, providing a valuable resource for further studies. Our results also uncovered a diverse set of novel phenotypes including nuclear enlargement in all glomerular cell types in aging and disease, as well as an aging-related pattern of regional thickening of the Bowman's capsule basement membrane near the tubular-glomerular junction.
传统上,人们通过微米级光学显微镜来研究肾小体的整体生理学或分子分布,通过纳米级电子显微镜来研究薄切片的超微结构。虽然这些方法很强大,但它们在获取肾小体作为整体三维功能单元的详细视图方面存在局限性。为了填补这一知识空白,我们开发了一种新颖的流程,使用超分辨率荧光显微镜以100纳米分辨率对整个小鼠肾小体进行成像、重建和分析。该流程整合了手动和机器学习方法来注释和分析肾小球结构。使用这种方法,我们创建了18个详细的肾小球模型,这些模型来自一系列健康、衰老和患病模型小鼠,勾勒出了所有主要结构和细胞类型。这些模型已在一个在线存储库中公开提供,为进一步研究提供了宝贵资源。我们的结果还发现了一系列新的表型,包括衰老和疾病中所有肾小球细胞类型的核增大,以及肾小管-肾小球连接处附近鲍曼囊基底膜的衰老相关区域增厚模式。
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