title: Ret and Etv4 Promote Directed Movements of Progenitor Cells during Renal Branching Morphogenesis.
abstract: Branching morphogenesis of the epithelial ureteric bud forms the renal collecting duct system and is critical for normal nephron number, while low nephron number is implicated in hypertension and renal disease. Ureteric bud growth and branching requires GDNF signaling from the surrounding mesenchyme to cells at the ureteric bud tips, via the Ret receptor tyrosine kinase and coreceptor Gfrα1; Ret signaling up-regulates transcription factors Etv4 and Etv5, which are also critical for branching. Despite extensive knowledge of the genetic control of these events, it is not understood, at the cellular level, how renal branching morphogenesis is achieved or how Ret signaling influences epithelial cell behaviors to promote this process. Analysis of chimeric embryos previously suggested a role for Ret signaling in promoting cell rearrangements in the nephric duct, but this method was unsuited to study individual cell behaviors during ureteric bud branching. Here, we use Mosaic Analysis with Double Markers (MADM), combined with organ culture and time-lapse imaging, to trace the movements and divisions of individual ureteric bud tip cells. We first examine wild-type clones and then Ret or Etv4 mutant/wild-type clones in which the mutant and wild-type sister cells are differentially and heritably marked by green and red fluorescent proteins. We find that, in normal kidneys, most individual tip cells behave as self-renewing progenitors, some of whose progeny remain at the tips while others populate the growing UB trunks. In Ret or Etv4 MADM clones, the wild-type cells generated at a UB tip are much more likely to remain at, or move to, the new tips during branching and elongation, while their Ret-/- or Etv4-/- sister cells tend to lag behind and contribute only to the trunks. By tracking successive mitoses in a cell lineage, we find that Ret signaling has little effect on proliferation, in contrast to its effects on cell movement. Our results show that Ret/Etv4 signaling promotes directed cell movements in the ureteric bud tips, and suggest a model in which these cell movements mediate branching morphogenesis.
authors.affiliation: Department of Genetics and Development, Columbia University, New York, New York, United States of America.
authors.forename: Paul
authors.initials: P
authors.lastname: Riccio
authors.affiliation: Department of Genetics and Development, Columbia University, New York, New York, United States of America.
authors.forename: Cristina
authors.initials: C
authors.lastname: Cebrian
authors.affiliation: Department of Microbiology, Immunology, and Cancer Biology, University of Virginia School of Medicine, Charlottesville, Virginia, United States of America.|Center for Brain Immunology and Glia, University of Virginia School of Medicine, Charlottesville, Virginia, United States of America.
authors.forename: Hui
authors.initials: H
authors.lastname: Zong
authors.affiliation: Developmental Neurobiology, IST Austria (Institute of Science and Technology Austria), Klosterneuburg, Austria.
authors.forename: Simon
authors.initials: S
authors.lastname: Hippenmeyer
authors.affiliation: Department of Genetics and Development, Columbia University, New York, New York, United States of America.
authors.forename: Frank
authors.initials: F
authors.lastname: Costantini
mesh_terms: D000818:Animals; D002465:Cell Movement; D005260:Female; D007668:Kidney / Q000196:embryology*; D008297:Male; D051379:Mice; D009024:Morphogenesis; D009924:Organ Culture Techniques; D050783:Proto-Oncogene Proteins c-ets / Q000502:physiology*; D051096:Proto-Oncogene Proteins c-ret / Q000502:physiology*; D013234:Stem Cells / Q000502:physiology*
chemical_list: C540571:Etv4 protein, mouse; D050783:Proto-Oncogene Proteins c-ets; D051096:Proto-Oncogene Proteins c-ret; C494999:Ret protein, mouse