Glomerular endothelial cell maturation depends on ADAM10, a key regulator of Notch signaling
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The principal function of glomeruli is to filter blood through a highly specialized filtration barrier consisting of a fenestrated endothelium, the glomerular basement membrane and podocyte foot processes. Previous studies have uncovered a crucial role of endothelial a disintegrin and metalloprotease 10 (ADAM10) and Notch signaling in the development of glomeruli, yet the resulting defects have not been further characterized nor understood in the context of kidney development. Here, we used several different experimental approaches to analyze the kidneys and glomeruli from mice lacking ADAM10 in endothelial cells (A10ΔEC mice). Scanning electron microscopy of glomerular casts demonstrated enlarged vascular diameter and increased intussusceptive events in A10ΔEC glomeruli compared to controls. Consistent with these findings, genes known to regulate vessel caliber (Apln, AplnR and Vegfr3) are significantly upregulated in A10ΔEC glomeruli. Moreover, transmission electron microscopy revealed the persistence of diaphragms in the fenestrae of A10ΔEC glomerular endothelial cells, which was corroborated by the elevated expression of the protein PLVAP/PV-1, an integral component of fenestral diaphragms. Analysis of gross renal vasculature by light sheet microscopy showed no major alteration of the branching pattern, indicating a localized importance of ADAM10 in the glomerular endothelium. Since intussusceptions and fenestrae with diaphragms are normally found in developing, but not mature glomeruli, our results provide the first evidence for a crucial role of endothelial ADAM10, a key regulator of Notch signaling, in promoting the development and maturation of the glomerular vasculature.
KeywordsGlomeruli Endothelial cells A disintegrin and metalloprotease 10 (ADAM10) Notch Fenestra Diaphragms
G. Farber is currently supported by a predoctoral Fellowship from the American Heart Association and was previously supported by Molecular and Cellular Biology T32 training Grant from the National Institutes of Health, 5T32GM008539. These studies were supported in part by the National Institutes of Health R01 Grant GM64750 to CPB. We would like to thank Dr. Alison North and Rockefeller University’s Bio-Imaging Resource Center for the training and usage of the light sheet microscopy and image analysis, Dr. Kunihiro Uryu and Rockefeller University’s Electron Microscopy Resource Center for training on and usage of scanning electron microscopy. A special thanks to Lee Cohen Gould and Juan Jimenez at the Weill Cornell Medicine Imaging Core Facility for the preparation of the transmission electron microscopy samples and training, and Dr. Katia Manova, Ning Fan and Afsar Barlas from the Molecular Cytology Core Facility at Memorial Sloan-Kettering Cancer Center (supported by the Cancer Center Support Grant P30CA008748). S. Monette and the Laboratory of Comparative Pathology are also supported in part by Cancer Center Support Grant P30CA008748.
GF and CB conceived of this study, RH, SL, SM, JM and CMS performed experiments and interpreted the results, GF drafted the manuscript, all authors contributed to editing, and GF and RPS prepared the figures.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
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