title: Distinct Requirements for Vacuolar Protein Sorting 34 Downstream Effector Phosphatidylinositol 3-Phosphate 5-Kinase in Podocytes Versus Proximal Tubular Cells.
abstract: The mechanisms by which the glomerular filtration barrier prevents the loss of large macromolecules and simultaneously, maintains the filter remain poorly understood. Recent studies proposed that podocytes have an active role in both the endocytosis of filtered macromolecules and the maintenance of the filtration barrier. Deletion of a key endosomal trafficking regulator, the class 3 phosphatidylinositol (PtdIns) 3-kinase vacuolar protein sorting 34 (Vps34), in podocytes results in aberrant endosomal membrane morphology and podocyte dysfunction. We recently showed that the vacuolation phenotype in cultured Vps34-deficient podocytes is caused by the absence of a substrate for the Vps34 downstream effector PtdIns 3-phosphate 5-kinase (PIKfyve), which phosphorylates Vps34-generated PtdIns(3)P to produce PtdIns (3,5)P2. PIKfyve perturbation and PtdIns(3,5)P2 reduction result in massive membrane vacuolation along the endosomal system, but the cell-specific functions of PIKfyve in vivo remain unclear. We show here that the genetic deletion of PIKfyve in endocytically active proximal tubular cells resulted in the development of large cytoplasmic vacuoles caused by arrested endocytic traffic progression at a late-endosome stage. In contrast, deletion of PIKfyve in glomerular podocytes did not significantly alter the endosomal morphology, even in age 18-month-old mice. However, on culturing, the PIKfyve-deleted podocytes developed massive cytoplasmic vacuoles. In summary, these data suggest that glomerular podocytes and proximal tubules have different requirements for PIKfyve function, likely related to distinct in vivo needs for endocytic flux.
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan; and.
authors.forename: Madhusudan
authors.initials: M
authors.lastname: Venkatareddy
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan; and.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Verma
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan; and.
authors.forename: Anne
authors.initials: A
authors.lastname: Kalinowski
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan; and.
authors.forename: Sanjeevkumar R
authors.initials: SR
authors.lastname: Patel
authors.affiliation: Department of Physiology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Assia
authors.initials: A
authors.lastname: Shisheva
authors.affiliation: Division of Nephrology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan; and pungarg@umich.edu.
authors.forename: Puneet
authors.initials: P
authors.lastname: Garg
mesh_terms: D000818:Animals; D058546:Class III Phosphatidylinositol 3-Kinases / Q000502:physiology*; D011992:Endosomes; D007678:Kidney Glomerulus; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000201:enzymology*; D051379:Mice; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000502:physiology*; D050199:Podocytes / Q000201:enzymology*
chemical_list: D019869:Phosphatidylinositol 3-Kinases; D058546:Class III Phosphatidylinositol 3-Kinases; C116930:Pikfyve protein, mouse
keywords: glomerulus; podocyte; proteinuria
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