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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 and John F. Bertram
JASN October 2016, 27 (10) 3093-3104; DOI: https://doi.org/10.1681/ASN.2015121340
Victor G. Puelles
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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James W. van der Wolde
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Keith E. Schulze
‡Monash Micro Imaging, Monash University, Melbourne, Australia; and
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Kieran M. Short
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
§Biochemistry and Molecular Biology, and
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Milagros N. Wong
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Jonathan G. Bensley
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Luise A. Cullen-McEwen
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Georgina Caruana
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Stacey N. Hokke
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Jinhua Li
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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Stephen D. Firth
‡Monash Micro Imaging, Monash University, Melbourne, Australia; and
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Ian S. Harper
‡Monash Micro Imaging, Monash University, Melbourne, Australia; and
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David J. Nikolic-Paterson
‖Medicine,
¶Department of Nephrology, Monash Health, Melbourne, Australia
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John F. Bertram
Departments of *Anatomy and Developmental Biology,
†Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and
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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.

  • podocyte
  • renal morphology
  • glomerular disease
  • Copyright © 2016 by the American Society of Nephrology
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Journal of the American Society of Nephrology: 27 (10)
Journal of the American Society of Nephrology
Vol. 27, Issue 10
October 2016
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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
JASN Oct 2016, 27 (10) 3093-3104; DOI: 10.1681/ASN.2015121340

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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
JASN Oct 2016, 27 (10) 3093-3104; DOI: 10.1681/ASN.2015121340
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More in this TOC Section

  • Dapagliflozin Prevents Kidney Glycogen Accumulation and Improves Renal Proximal Tubule Cell Functions in a Mouse Model of Glycogen Storage Disease Type 1b
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  • glomerular disease

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