Real-time analysis of ureteric bud branching morphogenesis in vitro.
While it is clear that the normal branching morphogenesis of the ureteric bud (UB) is critical for development of the metanephric kidney, the specific patterns of branching and growth have heretofore only been inferred from static images. Here, we present a systematic time-lapse analysis of UB branching morphogenesis during the early development of the mouse kidney in organ culture. Metanephric primordia from Hoxb7/GFP transgenic embryos were cultured for 3-4 days, and GFP images of the UB taken every 30 min were assembled into movies. Analysis of these movies (available as )revealed that the UB is a highly plastic structure, which can branch in a variety of complex patterns, including terminal bifid, terminal trifid, and lateral branching. To examine kinetic parameters of branching and elongation, skeletal representations of the UB were used to measure the number of segments and branch points and the length of each segment as a function of time and of branch generation. These measurements provide a baseline for future studies on mutant kidneys with defects in renal development. To illustrate how these quantitative methods can be applied to the analysis of abnormal kidney development, we examined the effects of the MEK1 inhibitor PD98059 on renal organ cultures and confirmed a previous report that the drug has a specific inhibitory effect on UB branching as opposed to elongation.
Watanabe
Tomoko
T
Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
Costantini
Frank
F
D000818:Animals; D001837:Body Weights and Measures; D003952:Diagnostic Imaging; D005419:Flavonoids / Q000494:pharmacology*; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D007668:Kidney / Q000187:drug effects* / Q000196:embryology*; D008164:Luminescent Proteins; D048369:MAP Kinase Kinase 1; D051379:Mice / Q000196:embryology*; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors; D009024:Morphogenesis; D009924:Organ Culture Techniques; D013997:Time Factors
D005419:Flavonoids; D018398:Homeodomain Proteins; C093253:Hoxb7 protein, mouse; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D048369:MAP Kinase Kinase 1; C482127:Map2k1 protein, mouse; D020929:Mitogen-Activated Protein Kinase Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one