title: Mosaic analysis of cell rearrangements during ureteric bud branching in dissociated/reaggregated kidney cultures and in vivo.
abstract: BACKGROUND
  Cell rearrangements mediated by GDNF/Ret signaling underlie the formation of the ureteric bud (UB) tip domain during kidney development. Whether FGF signaling also influences these rearrangements is unknown. Chimeric embryos are a powerful tool for examining the genetic controls of cellular behaviors, but generating chimeras by traditional methods is expensive and laborious. Dissociated fetal kidney cells can reorganize to form complex structures including branching UB tubules, providing an easier method to generate renal chimeras.
  
  
  RESULTS
  Cell behaviors in normal or chimeric kidney cultures were investigated using time-lapse imaging. In Spry1(-/-) ↔ wild-type chimeras, cells lacking Spry1 (a negative regulator of Ret and FGF receptor signaling) preferentially occupied the UB tips, as previously observed in traditional chimeras, thus validating this experimental system. In Fgfr2(UB-/-) ↔ wild-type chimeras, the wild-type cells preferentially occupied the tips. Independent evidence for a role of Fgfr2 in UB tip formation was obtained using Mosaic mutant Analysis with Spatial and Temporal control of Recombination (MASTR).
  
  
  CONCLUSIONS
  Dissociation and reaggregation of fetal kidney cells of different genotypes, with suitable fluorescent markers, provides an efficient way to analyze cell behaviors in chimeric cultures. FGF/Fgfr2 signaling promotes UB cell rearrangements that form the tip domain, similarly to GDNF/Ret signaling.
authors.affiliation: Department of Genetics and Development, Columbia University Medical Center, New York, NY.
authors.forename: Kevin
authors.initials: K
authors.lastname: Leclerc
authors.affiliation: Department of Genetics and Development, Columbia University Medical Center, New York, NY.
authors.forename: Frank
authors.initials: F
authors.lastname: Costantini
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D007668:Kidney / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D015398:Signal Transduction / Q000502:physiology*
keywords: FGF receptor 2; MASTR; Sprouty1; chimera; receptor tyrosine kinase
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