title: Tracking the fate of glomerular epithelial cells in vivo using serial multiphoton imaging in new mouse models with fluorescent lineage tags.
abstract: Podocytes are critical in the maintenance of a healthy glomerular filter; however, they have been difficult to study in the intact kidney because of technical limitations. Here we report the development of serial multiphoton microscopy (MPM) of the same glomeruli over several days to visualize the motility of podocytes and parietal epithelial cells (PECs) in vivo. In podocin-GFP mice, podocytes formed sporadic multicellular clusters after unilateral ureteral ligation and migrated into the parietal Bowman's capsule. The tracking of single cells in podocin-confetti mice featuring cell-specific expression of CFP, GFP, YFP or RFP revealed the simultaneous migration of multiple podocytes. In phosphoenolpyruvate carboxykinase (PEPCK)-GFP mice, serial MPM found PEC-to-podocyte migration and nanotubule connections. Our data support a highly dynamic rather than a static nature of the glomerular environment and cellular composition. Future application of this new approach should advance our understanding of the mechanisms of glomerular injury and regeneration.
authors.affiliation: 1] Department of Physiology and Biophysics, Zilkha Neurogenetic Institute, University of Southern California, Los Angeles, California, USA. [2] Department of Medicine, University of Southern California, Los Angeles, California, USA. [3] Department II of Internal Medicine and Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany.
authors.forename: Matthias J
authors.initials: MJ
authors.lastname: Hackl
authors.forename: James L
authors.initials: JL
authors.lastname: Burford
authors.forename: Karie
authors.initials: K
authors.lastname: Villanueva
authors.forename: Lisa
authors.initials: L
authors.lastname: Lam
authors.forename: Katalin
authors.initials: K
authors.lastname: Suszták
authors.forename: Bernhard
authors.initials: B
authors.lastname: Schermer
authors.forename: Thomas
authors.initials: T
authors.lastname: Benzing
authors.forename: János
authors.initials: J
authors.lastname: Peti-Peterdi
mesh_terms: D000818:Animals; D019070:Cell Lineage / Q000502:physiology*; D002465:Cell Movement; D058948:Cell Tracking / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D007678:Kidney Glomerulus / Q000166:cytology* / Q000502:physiology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton; D023421:Models, Animal; D009928:Organ Specificity / Q000235:genetics; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics
chemical_list: D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; D049452:Green Fluorescent Proteins; D010729:Phosphoenolpyruvate Carboxykinase (GTP)