The ureter develops out of the ureteric bud (UB), but most work on the UB to date has focused on branching morphogenesis as it relates to development of the kidney collecting ducts. Little is known about the genes specifying ureter formation out of the UB. This is an important question because, as we show, the developing ureter, though non-branching in vivo, has the capacity to branch in vitro in a manner similar to the portion of the UB destined to undergo branching morphogenesis as it develops into the kidney collecting ducts. Based on extensive preliminary data (Figs. 1-39), we hypothesize that two major steps are involved in ureter formation: 1) outgrowth of the UB from the Wolffian duct (a step in common with the kidney collecting ducts) (SA1); and 2) a branch-inhibitory influence on a subpopulation of UB cells that results in the formation of a non-branching ureter (SA2). The goal of this proposal is to employ in vitro assays, many of which have been developed by our group (e.g. isolated UB culture, isolated ureter culture, non-branching 1MCD tubulogenesis assay) together with a demonstrated ability to perform large scale gene expression (array) analysis on small quantities of DNA, to identify specific subsets of morphogenetic genes critical for: 1) outgrowth of the UB from the Wolffian duct (SA1) and 2) the formation of a non-branched ureter out of the UB (SA2). Those genes that are expressed in an appropriate spatiotemporal manner for a role in ureter development (immunocytochemistry, in situ hybridization) will be functionally perturbed in the in the appropriate vitro system (e.g. isolated Wolffian duct culture, isolated UB culture, isolated ureter culture, branching and non-branching IMCD tubulogenesis assay). This will be done using a variety of techniques (neutralizing antibodies, antisense/morpholinos/RNAi, Specific inhibitors or activators, transfections). We have published experience with nearly all these techniques, and considerable data is presented to indicate the feasibility of the approach and our ability to perform sophisticated computational analyses of large sets of genes such as those that will be obtained in the proposed array experiments. Together, these studies should lead to the identification of genes that are critical for UB formation and that determine the fate of cells in the UB destined to form the ureter. (This proposal is specifically responsive to Program Announcement PAR-02- 143).

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK065831-04
Application #
7154796
Study Section
Special Emphasis Panel (ZRG1-UROL (52))
Program Officer
Hoshizaki, Deborah K
Project Start
2004-01-01
Project End
2008-11-30
Budget Start
2006-12-01
Budget End
2008-11-30
Support Year
4
Fiscal Year
2007
Total Cost
$354,544
Indirect Cost
Name
University of California San Diego
Department
Pediatrics
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
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Tee, James B; Choi, Yohan; Shah, Mita M et al. (2010) Protein kinase A regulates GDNF/RET-dependent but not GDNF/Ret-independent ureteric bud outgrowth from the Wolffian duct. Dev Biol 347:337-47
Shah, Mita M; Sakurai, Hiroyuki; Sweeney, Derina E et al. (2010) Hs2st mediated kidney mesenchyme induction regulates early ureteric bud branching. Dev Biol 339:354-65
Choi, Yohan; Tee, James B; Gallegos, Thomas F et al. (2009) Neuropeptide Y functions as a facilitator of GDNF-induced budding of the Wolffian duct. Development 136:4213-24
Shah, Mita M; Tee, James B; Meyer, Tobias et al. (2009) The instructive role of metanephric mesenchyme in ureteric bud patterning, sculpting, and maturation and its potential ability to buffer ureteric bud branching defects. Am J Physiol Renal Physiol 297:F1330-41
Nigam, Sanjay K; Shah, Mita M (2009) How does the ureteric bud branch? J Am Soc Nephrol 20:1465-9
Wu, Wei; Kitamura, Shinji; Truong, David M et al. (2009) Beta1-integrin is required for kidney collecting duct morphogenesis and maintenance of renal function. Am J Physiol Renal Physiol 297:F210-7

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