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

Victor G. Puelles, James W. van der Wolde, Keith E. Schulze, Kieran M. Short, Milagros N. Wong, Jonathan G. Bensley, Luise A. Cullen-McEwen, Georgina Caruana, Stacey N. Hokke, Jinhua Li, Stephen D. Firth, Ian S. Harper, David J. Nikolic-Paterson, John F. Bertram

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20 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)3093-3104
Number of pages12
JournalJournal of the American Society of Nephrology
Volume27
Issue number10
DOIs
Publication statusPublished - 1 Oct 2016

Keywords

  • podocyte
  • renal morphology
  • glomerular disease

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