title: Validation of a Three-Dimensional Method for Counting and Sizing Podocytes in Whole Glomeruli.
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.
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and victor.puellesrodriguez@monash.edu.
authors.forename: Victor G
authors.initials: VG
authors.lastname: Puelles
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: James W
authors.initials: JW
authors.lastname: van der Wolde
authors.affiliation: Monash Micro Imaging, Monash University, Melbourne, Australia; and.
authors.forename: Keith E
authors.initials: KE
authors.lastname: Schulze
authors.affiliation: Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and Biochemistry and Molecular Biology, and.
authors.forename: Kieran M
authors.initials: KM
authors.lastname: Short
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: Milagros N
authors.initials: MN
authors.lastname: Wong
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: Jonathan G
authors.initials: JG
authors.lastname: Bensley
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: Luise A
authors.initials: LA
authors.lastname: Cullen-McEwen
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: Georgina
authors.initials: G
authors.lastname: Caruana
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: Stacey N
authors.initials: SN
authors.lastname: Hokke
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Li
authors.affiliation: Monash Micro Imaging, Monash University, Melbourne, Australia; and.
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Firth
authors.affiliation: Monash Micro Imaging, Monash University, Melbourne, Australia; and.
authors.forename: Ian S
authors.initials: IS
authors.lastname: Harper
authors.affiliation: Medicine, Department of Nephrology, Monash Health, Melbourne, Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Nikolic-Paterson
authors.affiliation: Departments of Anatomy and Developmental Biology, Development and Stem Cells Program, Monash Biomedicine Discovery Institute, and.
authors.forename: John F
authors.initials: JF
authors.lastname: Bertram
mesh_terms: D000818:Animals; D002452:Cell Count / Q000379:methods*; D048429:Cell Size; D021621:Imaging, Three-Dimensional; D007678:Kidney Glomerulus / Q000166:cytology*; D051379:Mice; D050199:Podocytes / Q000166:cytology*
keywords: glomerular disease; podocyte; renal morphology