DEVELOPMENT OF THE URINARY COLLECTING SYSTEM

尿液收集系统的开发

基本信息

批准号：
6624897
负责人：
SANJAY K NIGAM
金额：
$ 28.59万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-09-01 至 2004-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6624897
关键词：
antisense nucleic acid enzyme mechanism gene expression histogenesis immunocytochemistry in situ hybridization laboratory mouse mesenchyme molecular cloning northern blottings nucleic acid sequence protein sequence protein tyrosine kinase renal tubule ureter urinary tract

项目摘要

The goal of this COMPETITIVE RENEWAL application is to provide a molecular explanation of the role of soluble factors and mesenchymal contact in development of the urinary collecting system from the ureteric bud (UB). The UB undergoes repetitive branching tubulogenesis during formation of the collecting system. In the previous project period, we developed two novel systems to study branching tubulogenesis of the UB: l) an in vitro UB cell culture model for branching tubulogenesis in which UB cell morphogenesis is stimulated by soluble factors produced from a metanephric mesenchymal cell line; and 2) a system to induce branching tubulogenesis in isolated UB tissue in the presence of soluble factors (which can be extended to study UB-mesenchyme interactions by recombining the cultured UB with mesenchyme). These are powerful systems in which to examine the function of genes involved in collecting system development. For reasons detailed in the proposal, novel and known receptor tyrosine kinases (RTKs) are likely to play a key role in both early and late UB morphogenesis. Using a method pioneered by us known as Codon Optimized Differential Display (CODD),, we targeted RTKs that might be involved in UB branching tubulogenesis. In addition to identifying at least 10 novel gene sequences, some of which are likely to be novel RTKs, we obtained the surprising result that more that 80% of the known RTKs upregulated during UB cell branching tubulogenesis belong to the Eph subfamily. Based on extensive preliminary data, we hypothesize that: l) As yet unexamined or new RTKs are involved in branching tubulogenesis of the UB specifically mediated by soluble factors (SA2); 2) Eph RTKs and their ephrin ligands play a role in branch elongation and guidance induced by direct contact of UB tips with the metanephric mesenchyme (SA1). Thus, we postulate (and provide data in support of) separate roles of soluble factors (UB branching morphogenesis) and mesenchymal contact (UB branch elongation and guidance) in collecting system development and propose that these are mediated by separate subfamilies of RTKs. We aim to characterize their: 1) spatiotemporal patterns of expression during development of the ureter and collecting ducts (Northerns, in situ hybridization, immunohistochemistry); 2) clone and sequence any novel RTKs of interest; and 3) perform a careful functional analysis of the role of these genes (using soluble monomeric ephrins and Ephs, antisense technology) in the unique model systems we established during the previous project period.

这种竞争性更新应用的目的是提供分子解释可溶性因子的作用和间质接触从输尿管芽（UB）开发尿收集系统中的作用。 UB在收集系统形成期间经历重复的分支小管发生。在上一个项目期间，我们开发了两个新型系统，研究了UB的分支小管发生：l）一种体外UB细胞培养模型，用于分支微管发生，其中跨eNaphephric间质细胞系产生的可溶性因子刺激了UB细胞的形态发生； 2）在存在可溶性因子的情况下诱导分离的UB组织中分支小管发生的系统（可以扩展到通过将培养的UB与间充质重新组合来研究UB - 间质相互作用）。这些是检查收集系统开发中涉及的基因功能的强大系统。出于提案中详细介绍的原因，新颖和已知的受体酪氨酸激酶（RTK）可能在UB形态发生中都起着关键作用。使用我们被称为密码子优化差异显示（CODD）的一种方法，我们靶向可能参与UB分支小管的RTK。除了鉴定至少10个新型基因序列（其中一些可能是新型的RTK）外，我们获得了令人惊讶的结果，即在UB细胞分支小管中上调的80％已知RTK属于EPH亚科。基于广泛的初步数据，我们假设：l）尚未审查或新RTK参与了由可溶性因子专门介导的UB的分支小管发生（SA2）； 2）EPH RTK及其Ephrin配体在分支伸长和指导中发挥作用，该分支和指导与UB TIPS与Metanephric间质的直接接触（SA1）引起。因此，我们假设（并提供支持）可溶性因子（UB分支形态发生）和间充质接触（UB分支伸长和指导）在收集系统开发中的单独作用（UB分支分支），并提出这些作用是由RTKS的单独亚科介导的。我们旨在表征它们：1）输尿管发育过程中表达的时空模式和收集管道（北部，原位杂交，免疫组织化学）； 2）克隆并序列任何有趣的新型RTK； 3）对我们在上一个项目期间建立的独特模型系统中这些基因的作用（使用可溶性单体晶状体和EPHS，反义技术）进行仔细的功能分析。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Expression of c-ret promotes morphogenesis and cell survival in mIMCD-3 cells.

c-ret 的表达促进 mIMCD-3 细胞的形态发生和细胞存活。

DOI：
10.1152/ajprenal.1999.276.4.f581
发表时间：
1999
期刊：
The American journal of physiology
影响因子：
0
作者：
O'Rourke,DA;Sakurai,H;Spokes,K;Kjelsberg,C;Takahashi,M;Nigam,S;Cantley,L
通讯作者：
Cantley,L

Regulation of ureteric bud branching morphogenesis by sulfated proteoglycans in the developing kidney.

DOI：
10.1016/j.ydbio.2004.04.029
发表时间：
2004-08
期刊：
Developmental biology
影响因子：
2.7
作者：
Dylan L. Steer;M. M. Shah-M.;K. Bush;R. O. Stuart;R. Sampogna;T. Meyer;C. Schwesinger;X. Bai;J. Esko;S. Nigam
通讯作者：
Dylan L. Steer;M. M. Shah-M.;K. Bush;R. O. Stuart;R. Sampogna;T. Meyer;C. Schwesinger;X. Bai;J. Esko;S. Nigam

Matrix metalloproteinases and their inhibitors regulate in vitro ureteric bud branching morphogenesis.

基质金属蛋白酶及其抑制剂调节体外输尿管芽分支形态发生。