title: Three-dimensional electron microscopy reveals the evolution of glomerular barrier injury.
abstract: Glomeruli are highly sophisticated filters and glomerular disease is the leading cause of kidney failure. Morphological change in glomerular podocytes and the underlying basement membrane are frequently observed in disease, irrespective of the underlying molecular etiology. Standard electron microscopy techniques have enabled the identification and classification of glomerular diseases based on two-dimensional information, however complex three-dimensional ultrastructural relationships between cells and their extracellular matrix cannot be easily resolved with this approach. We employed serial block face-scanning electron microscopy to investigate Alport syndrome, the commonest monogenic glomerular disease, and compared findings to other genetic mouse models of glomerular disease (Myo1e-/-, Ptpro-/-). These analyses revealed the evolution of basement membrane and cellular defects through the progression of glomerular injury. Specifically we identified sub-podocyte expansions of the basement membrane with both cellular and matrix gene defects and found a corresponding reduction in podocyte foot process number. Furthermore, we discovered novel podocyte protrusions invading into the glomerular basement membrane in disease and these occurred frequently in expanded regions of basement membrane. These findings provide new insights into mechanisms of glomerular barrier dysfunction and suggest that common cell-matrix-adhesion pathways are involved in the progression of disease regardless of the primary insult.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, UK.|Institute of Human Development, Faculty of Medical &Human Sciences, University of Manchester, UK.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Randles
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, UK.
authors.forename: Sophie
authors.initials: S
authors.lastname: Collinson
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, UK.|Electron Microscopy Core Facility, Faculty of Life Sciences, University of Manchester, UK.
authors.forename: Tobias
authors.initials: T
authors.lastname: Starborg
authors.affiliation: Electron Microscopy Core Facility, Faculty of Life Sciences, University of Manchester, UK.
authors.forename: Aleksandr
authors.initials: A
authors.lastname: Mironov
authors.affiliation: Department of Cell and Developmental Biology, SUNY Upstate Medical University, Syracuse, New York, USA.
authors.forename: Mira
authors.initials: M
authors.lastname: Krendel
authors.affiliation: Department of Nephrology, Medical School, Heinrich Heine University, Düsseldorf, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Königshausen
authors.affiliation: Department of Nephrology, Medical School, Heinrich Heine University, Düsseldorf, Germany.
authors.forename: Lorenz
authors.initials: L
authors.lastname: Sellin
authors.affiliation: Department of Cellular Pathology, John Radcliffe Hospital, Oxford, UK.
authors.forename: Ian S D
authors.initials: IS
authors.lastname: Roberts
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, UK.
authors.forename: Karl E
authors.initials: KE
authors.lastname: Kadler
authors.affiliation: Division of Nephrology, Washington University School of Medicine, St Louis, Missouri, USA.
authors.forename: Jeffrey H
authors.initials: JH
authors.lastname: Miner
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, UK.|Institute of Human Development, Faculty of Medical &Human Sciences, University of Manchester, UK.
authors.forename: Rachel
authors.initials: R
authors.lastname: Lennon
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D055786:Gene Knockout Techniques; D050533:Glomerular Basement Membrane / Q000000981:diagnostic imaging; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods; D007674:Kidney Diseases / Q000000981:diagnostic imaging* / Q000235:genetics; D051379:Mice; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D024461:Myosin Type I; D009218:Myosins / Q000235:genetics; D009394:Nephritis, Hereditary / Q000000981:diagnostic imaging* / Q000209:etiology; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3 / Q000235:genetics
chemical_list: C516890:Ptpro protein, mouse; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3; C505821:Myo1e protein, mouse; D024461:Myosin Type I; D009218:Myosins