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

doi:10.1681/ASN.2015121340

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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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 language	English
Pages (from-to)	3093-3104
Number of pages	12
Journal	Journal of the American Society of Nephrology
Volume	27
Issue number	10
DOIs	https://doi.org/10.1681/ASN.2015121340
Publication status	Published - 1 Oct 2016

Keywords

podocyte
renal morphology
glomerular disease

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10.1681/ASN.2015121340

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Puelles, V. G., van der Wolde, J. W., Schulze, K. E., Short, K. M., Wong, M. N., Bensley, J. G., Cullen-McEwen, L. A., Caruana, G., Hokke, S. N., Li, J., Firth, S. D., Harper, I. S., Nikolic-Paterson, D. J., & Bertram, J. F. (2016). Validation of a Three-Dimensional Method for Counting and Sizing Podocytes in Whole Glomeruli. Journal of the American Society of Nephrology, 27(10), 3093-3104. https://doi.org/10.1681/ASN.2015121340

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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.",

keywords = "podocyte, renal morphology, glomerular disease",

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

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doi = "10.1681/ASN.2015121340",

language = "English",

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journal = "Journal of the American Society of Nephrology",

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Puelles, VG, van der Wolde, JW, Schulze, KE, Short, KM, Wong, MN, Bensley, JG, Cullen-McEwen, LA, Caruana, G, Hokke, SN, Li, J, Firth, SD, Harper, IS, Nikolic-Paterson, DJ & Bertram, JF 2016, 'Validation of a Three-Dimensional Method for Counting and Sizing Podocytes in Whole Glomeruli', Journal of the American Society of Nephrology, vol. 27, no. 10, pp. 3093-3104. https://doi.org/10.1681/ASN.2015121340

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AU - Puelles, Victor G.

AU - van der Wolde, James W.

AU - Schulze, Keith E.

AU - Short, Kieran M.

AU - Wong, Milagros N.

AU - Bensley, Jonathan G.

AU - Cullen-McEwen, Luise A.

AU - Caruana, Georgina

AU - Hokke, Stacey N.

AU - Li, Jinhua

AU - Firth, Stephen D.

AU - Harper, Ian S.

AU - Nikolic-Paterson, David J.

AU - Bertram, John F.

PY - 2016/10/1

Y1 - 2016/10/1

N2 - 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.

AB - 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.

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KW - renal morphology

KW - glomerular disease

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SN - 1046-6673

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JO - Journal of the American Society of Nephrology

JF - Journal of the American Society of Nephrology

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

Abstract

Keywords

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Other files and links

Human podocyte depletion, glomerular hypertrophy and glomerulosclerosis

Histology Platform

Cite this

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

Abstract

Keywords

Access to Document

Other files and links

Projects

Human podocyte depletion, glomerular hypertrophy and glomerulosclerosis

Equipment

Histology Platform

Cite this