title: Intravital and Kidney Slice Imaging of Podocyte Membrane Dynamics.
abstract: In glomerular disease, podocyte injury results in a dramatic change in cell morphology known as foot process effacement. Remodeling of the actin cytoskeleton through the activity of small GTPases was identified as a key mechanism in effacement, with increased membrane activity and motility in vitro However, whether podocytes are stationary or actively moving cells in vivo remains debated. Using intravital and kidney slice two-photon imaging of the three-dimensional structure of mouse podocytes, we found that uninjured podocytes remained nonmotile and maintained a canopy-shaped structure over time. On expression of constitutively active Rac1, however, podocytes changed shape by retracting processes and clearly exhibited domains of increased membrane activity. Constitutive activation of Rac1 also led to podocyte detachment from the glomerular basement membrane, and we detected detached podocytes crawling on the surface of the tubular epithelium and occasionally, in contact with peritubular capillaries. Podocyte membrane activity also increased in the inflammatory environment of immune complex-mediated GN. Our results provide evidence that podocytes transition from a static to a dynamic state in vivo, shedding new light on mechanisms in foot process effacement.
authors.affiliation: Department of Pathology and Immunology and.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Brähler
authors.affiliation: Department of Pathology and Immunology and.
authors.forename: Haiyang
authors.initials: H
authors.lastname: Yu
authors.affiliation: Department of Pathology and Immunology and.
authors.forename: Hani
authors.initials: H
authors.lastname: Suleiman
authors.affiliation: Department of Pathology and Immunology and.
authors.forename: Gokul M
authors.initials: GM
authors.lastname: Krishnan
authors.affiliation: Department of Pathology and Immunology and.
authors.forename: Brian T
authors.initials: BT
authors.lastname: Saunders
authors.affiliation: Kidney Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Jeffrey B
authors.initials: JB
authors.lastname: Kopp
authors.affiliation: Division of Nephrology, Washington University School of Medicine, St. Louis, Missouri; and.
authors.forename: Jeffrey H
authors.initials: JH
authors.lastname: Miner
authors.affiliation: Department of Pathology and Immunology and bzinselmeyer@path.wustl.edu shaw.andrey@gene.com.
authors.forename: Bernd H
authors.initials: BH
authors.lastname: Zinselmeyer
authors.affiliation: Department of Pathology and Immunology and bzinselmeyer@path.wustl.edu shaw.andrey@gene.com.
authors.forename: Andrey S
authors.initials: AS
authors.lastname: Shaw
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000502:physiology*; D000069416:Intravital Microscopy; D007668:Kidney / Q000166:cytology; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D050199:Podocytes / Q000502:physiology* / Q000648:ultrastructure*
keywords: cytoskeleton; glomerular disease; podocyte