Validation of a Three-Dimensional Method for Counting and Sizing Podocytes in Whole Glomeruli

Podocyte depletion is sufficient for the development of numerous glomerular diseases and can be absolute (loss of podocytes) or relative (reduced number of podocytes per volume of glomerulus). Commonly used methods to quantify podocyte depletion introduce bias, whereas gold standard stereologic meth...

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Veröffentlicht in:Journal of the American Society of Nephrology 2016-10, Vol.27 (10), p.3093-3104
Hauptverfasser: Puelles, Victor G, van der Wolde, James W, Schulze, Keith E, Short, Kieran M, Wong, Milagros N, Bensley, Jonathan G, Cullen-McEwen, Luise A, Caruana, Georgina, Hokke, Stacey N, Li, Jinhua, Firth, Stephen D, Harper, Ian S, Nikolic-Paterson, David J, Bertram, John F
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Sprache:eng
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Zusammenfassung:Podocyte depletion is sufficient for the development of numerous glomerular diseases and can be absolute (loss of podocytes) or relative (reduced number of podocytes per volume of glomerulus). Commonly used methods to quantify podocyte depletion introduce bias, whereas gold standard stereologic methodologies are time consuming and impractical. We developed a novel approach for assessing podocyte depletion in whole glomeruli that combines immunofluorescence, optical clearing, confocal microscopy, and three-dimensional analysis. We validated this method in a transgenic mouse model of selective podocyte depletion, in which we determined dose-dependent alterations in several quantitative indices of podocyte depletion. This new approach provides a quantitative tool for the comprehensive and time-efficient analysis of podocyte depletion in whole glomeruli.
ISSN:1046-6673
1533-3450
DOI:10.1681/asn.2015121340