Combined use of electron microscopy and intravital imaging captures morphological and functional features of podocyte detachment.
Combined use of electron microscopy and intravital imaging captures morphological and functional features of podocyte detachment.
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DOI：
10.1007/s00424-017-2020-0
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发表时间：
2017-08
期刊：
影响因子：
--
通讯作者：
Peti-Peterdi J
中科院分区：
文献类型：
--
作者：
Burford JL;Gyarmati G;Shirato I;Kriz W;Lemley KV;Peti-Peterdi J
由于实时研究肾小球滤过屏障（glomerular filtration barrier， GFB）的技术限制，各种肾小球病理中足细胞损伤和蛋白尿的发展尚不完全清楚。本研究采用透射电子显微镜（TEM）和体内多光子显微镜（MPM）相结合的方法，在大鼠嘌呤霉素氨基核苷（PAN）模型中直接观察局灶性节段性肾小球硬化（FSGS）发展过程中GFB的早期形态和功能变化。我们假设这种TEM + MPM联合实验方法将提供重大的技术改进，有助于我们对足细胞脱离的机制理解。雄性Sprague-Dawley（用于TEM）或Munich-Wistar-Frömter（用于MPM）大鼠给予单剂量100-150 mg/kg体重的PAN i.p.，并在给药后2-14天的不同时间处死，肾脏处理TEM或手术仪器用于体内MPM成像。两种技术都显示早在PAN术后2-3天就出现了传唤细胞间隙肥大和囊性扩张。第8-10天出现内脏上皮与鲍曼囊壁的粘连（粘连）。TEM提供了无与伦比的足突重塑分辨率，而MPM显示了假性囊肿填充、排空和破裂的快速动态，以及内皮细胞和足突细胞损伤、定向滤过和足突细胞脱落。由于TEM和MPM的互补优势，这种联合方法可以提供FSGS发展过程中足细胞形态和功能变化的异常全面和动态的描述。这些结果促进了我们对各种细胞类型、血流动力学和机械力在肾小球病理发展中的作用和重要性的理解。
The development of podocyte injury and albuminuria in various glomerular pathologies is still incompletely understood due to technical limitations in studying the glomerular filtration barrier (GFB) in real-time. We aimed to directly visualize the early morphological and functional changes of the GFB during the development of focal segmental glomerulosclerosis (FSGS) using a combination of transmission electron microscopy (TEM) and in vivo multiphoton microscopy (MPM) in the rat puromycin aminonucleoside (PAN) model. We hypothesized that this combined TEM + MPM experimental approach would provide a major technical improvement that would benefit our mechanistic understanding of podocyte detachment. Male Sprague-Dawley (for TEM) or Munich-Wistar-Frömter (for MPM) rats were given a single dose of 100–150 mg/kg body weight PAN i.p. and were either sacrificed and the kidneys processed for TEM or surgically instrumented for in vivo MPM imaging at various times 2–14 days after PAN administration. Both techniques demonstrated hypertrophy and cystic dilatations of the subpodocyte space that developed as early as 2–3 days after PAN. Adhesions of the visceral epithelium to the parietal Bowman’s capsule (synechiae) appeared at days 8–10. TEM provided unmatched resolution of podocyte foot process remodeling, while MPM revealed the rapid dynamics of pseudocyst filling, emptying, and rupture, as well as endothelial and podocyte injury, misdirected filtration, and podocyte shedding. Due to the complementary advantages of TEM and MPM, this combined approach can provide an unusally comprehensive and dynamic portrayal of the alterations in podocyte morphology and function during FSGS development. The results advance our understanding of the role and importance of the various cell types, hemodynamics, and mechanical forces in the development of glomerular pathology.
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影响因子：
5.2
作者：
Kriz W;Hähnel B;Hosser H;Rösener S;Waldherr R
通讯作者：
Waldherr R
DOI：
10.1152/ajprenal.00521.2005
发表时间：
2006-08-01
影响因子：
4.2
作者：
Kang, Jung Julie;Toma, Ildiko;Peti-Peterdi, Janos
通讯作者：
Peti-Peterdi, Janos
DOI：
10.1152/ajprenal.00272.2006
发表时间：
2007-02-01
影响因子：
4.2
作者：
Eyre, Jeanette;Ioannou, Kyriakos;Topham, Peter S.
通讯作者：
Topham, Peter S.
影响因子：
15.9
作者：
Burford, James L.;Villanueva, Kane;Peti-Peterdi, Janos
通讯作者：
Peti-Peterdi, Janos
影响因子：
19.6
作者：
Kim, YH;Goyal, M;Wiggins, R
通讯作者：
Wiggins, R