Novel 3D analysis using optical tissue clearing documents the evolution of murine rapidly progressive glomerulonephritis

Victor G Puelles; David Fleck; Lena Ortz; Stella Papadouri; Thiago Strieder; Alexander M C Böhner; James W van der Wolde; Michael Vogt; Turgay Saritas; Christoph Kuppe; Astrid Fuss; Sylvia Menzel; Barbara M Klinkhammer; Gerhard Müller-Newen; Felix Heymann; Leon Decker; Fabian Braun; Oliver Kretz; Tobias B Huber; Etsuo A Susaki; Hiroki R Ueda; Peter Boor; Jürgen Floege; Rafael Kramann; Christian Kurts; John F Bertram; Marc Spehr; David J Nikolic-Paterson; Marcus J Moeller

doi:10.1016/j.kint.2019.02.034

Novel 3D analysis using optical tissue clearing documents the evolution of murine rapidly progressive glomerulonephritis

Kidney Int. 2019 Aug;96(2):505-516. doi: 10.1016/j.kint.2019.02.034. Epub 2019 Mar 15.

Authors

Victor G Puelles¹, David Fleck², Lena Ortz³, Stella Papadouri³, Thiago Strieder³, Alexander M C Böhner⁴, James W van der Wolde⁵, Michael Vogt⁶, Turgay Saritas³, Christoph Kuppe³, Astrid Fuss³, Sylvia Menzel³, Barbara M Klinkhammer⁷, Gerhard Müller-Newen⁸, Felix Heymann⁹, Leon Decker³, Fabian Braun¹⁰, Oliver Kretz¹⁰, Tobias B Huber¹⁰, Etsuo A Susaki¹¹, Hiroki R Ueda¹², Peter Boor⁷, Jürgen Floege³, Rafael Kramann³, Christian Kurts⁴, John F Bertram⁵, Marc Spehr², David J Nikolic-Paterson¹³, Marcus J Moeller¹⁴

Affiliations

¹ Department of Nephrology and Clinical Immunology, RWTH Aachen University Clinic, Aachen, Germany; III. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Department of Nephrology, Monash Health, and Centre for Inflammatory Diseases, Monash University, Melbourne, Australia. Electronic address: [email protected].
² Department of Chemosensation, RWTH Aachen University, Aachen.
³ Department of Nephrology and Clinical Immunology, RWTH Aachen University Clinic, Aachen, Germany.
⁴ Institute of Experimental Immunology, Rheinische-Friedrichs-Wilhelms University of Bonn, Bonn, Germany.
⁵ Development and Stem Cells Program, Monash Biomedicine Discovery Institute and Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia.
⁶ Core Facility "Two-Photon Imaging" IZKF Aachen, RWTH University of Aachen, Aachen, Germany.
⁷ Department of Nephrology and Clinical Immunology, RWTH Aachen University Clinic, Aachen, Germany; Institute of Pathology, RWTH University of Aachen, Aachen, Germany.
⁸ Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Aachen, Germany.
⁹ Department of Gastroenterology, RWTH University of Aachen, Aachen, Germany.
¹⁰ III. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
¹¹ Department of Systems Pharmacology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; Laboratory for Synthetic Biology, RIKEN Quantitative Biology Center, Osaka, Japan; PRESTO, Japan Science and Technology Agency, Saitama, Japan.
¹² Department of Systems Pharmacology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; Laboratory for Synthetic Biology, RIKEN Quantitative Biology Center, Osaka, Japan.
¹³ Department of Nephrology, Monash Health, and Centre for Inflammatory Diseases, Monash University, Melbourne, Australia.
¹⁴ Department of Nephrology and Clinical Immunology, RWTH Aachen University Clinic, Aachen, Germany; Interdisciplinary Centre for Clinical Research (IZKF Aachen), RWTH Aachen University Hospital, Aachen, Germany; Heisenberg Chair for Preventive and Translational Nephrology, Division of Nephrology, RWTH Aachen University, Aachen, Germany. Electronic address: [email protected].

PMID: 31155155
DOI: 10.1016/j.kint.2019.02.034

Abstract

Recent developments in optical tissue clearing have been difficult to apply for the morphometric analysis of organs with high cellular content and small functional structures, such as the kidney. Here, we establish combinations of genetic and immuno-labelling for single cell identification, tissue clearing and subsequent de-clarification for histoimmunopathology and transmission electron microscopy. Using advanced light microscopy and computational analyses, we investigated a murine model of crescentic nephritis, an inflammatory kidney disease typified by immune-mediated damage to glomeruli leading to the formation of hypercellular lesions and the rapid loss of kidney function induced by nephrotoxic serum. Results show a graded susceptibility of the glomeruli, significant podocyte loss and capillary injury. These effects are associated with activation of parietal epithelial cells and formation of glomerular lesions that may evolve and obstruct the kidney tubule, thereby explaining the loss of kidney function. Thus, our work provides new high-throughput endpoints for the analysis of complex tissues with single-cell resolution.

Keywords: computational analysis; crescentic nephritis; optical clearing; parietal cell activation; podocyte loss.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Capillaries
Disease Models, Animal
Disease Progression
Fluorescence
Fluorescent Dyes / chemistry
Genes, Reporter / genetics
Glomerulonephritis / immunology
Glomerulonephritis / pathology*
Green Fluorescent Proteins / chemistry
Green Fluorescent Proteins / genetics
Histocytological Preparation Techniques / methods*
Humans
Imaging, Three-Dimensional*
Male
Mice
Mice, Transgenic
Microscopy, Electron, Transmission
Podocytes / physiology*
Podocytes / ultrastructure
Single-Cell Analysis / methods*

Substances

Fluorescent Dyes
enhanced green fluorescent protein
Green Fluorescent Proteins