Mechanisms of Renal Tubulogenesis

肾小管发生机制

• 批准号: 7988980
• 负责人: Keith E Mostov
• 金额: $ 6.92万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7988980
• 关键词: 3-Dimensional; 4-Hydroxy-Tamoxifen; Abbreviations; Acute Kidney Failure; Acute Kidney Tubular Necrosis; Affect; Binding; Biochemical; Biological Assay; Biological Models; Canis familiaris; Cell Culture Techniques; Cell Line; Cell surface; Cells; Cholera Toxin; Collagen; Complex; Conditioned Culture Media; Confocal Microscopy; Cyst; Defect; Development; Disease; Dominant-Negative Mutation; Doxycycline; Epithelial; Epithelial Cells; Estrogen Receptor 1; Extracellular Matrix; Family; Fibrosis; Four-dimensional; GTPase-Activating Proteins; Gel; Genes; Growth Factor; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Health; Hepatocyte Growth Factor; Hereditary Disease; Immunofluorescence Immunologic; Injury; Kidney; Kidney Diseases; Kinetics; Learning; MAP Kinase Gene; MAP Kinase Modules; Matrix Metalloproteinases; Membrane Microdomains; Mesenchymal; Microtubules; Mitogen-Activated Protein Kinases; Molecular; Myosin Light Chains; Natural regeneration; Organ; Pathway interactions; Phosphatidylinositols; Phosphotransferases; Physiological; Process; RNA Interference; Ras/Raf; Receptor Protein-Tyrosine Kinases; Recovery; Renal tubule structure; Research Personnel; Rho-associated kinase; Role; Signal Pathway; Signal Transduction; Staging; System; Testing; Thick; Tight Junctions; Time; Tube; Video Microscopy; Work; cell type; in vivo; kidney cell; meetings; methyl-beta-cyclodextrin; monolayer; nephrogenesis; phosphoinositide-3,4,5-triphosphate; phosphoinositide-3,4-bisphosphate; prevent; programs; rho; small hairpin RNA; stem; time use; two-dimensional

The kidney, like many epithelial organs, consists mainly of tubules lined by a monolayer of polarized epithelial cells. Though we have learned a great deal about the genes that control kidney development, we know much less about the mechanisms by which cells rearrange themselves into tubules. Weuse a simplified model system of Madin-Darbycanine kidney (MDCK) cells grown in a 3 dimensionalcollagen gel. Single MDCK cellsgrow to form hollow cysts lined by a monolayer of epithelial cells. When these cysts are stimulated with Hepatocyte Growth Factor (HGF), they form tubules over a period of ~3 days. This is a good model system for studying renal tubule formation, and can also provide informationon several pathophysiological processes, such as recovery from acute tubular necrosis, renal fibrosis, polycystickidney disease and perhaps renal regeneration from stem cells.We will study the signaling pathways by which cells respond to HGF. The kinetics of activation of ERKsuggest that it is activated by a MAPK cascadeinvolving B-Raf, rather than the better known Raf-1. We will test this hypothesis by studying activation of components of the B-Raf pathway and by using dominant negatives and RNAi. We will test the involvement of Rho family GTPases, Rho Kinase and lipid rafts in the early stages of tubulogenesis, especially the formation of extensions from the basolateral surface of cells in cysts. We will use time lapse confocal microscopy to observe the effects of perturbing these molecular components on extension formation.We will test the involvement of phosphatidyl inositol (3,4,5)P3 [PI(3,4,5)P3]in extension formation. We have found that exogenous PI(3,4,5)P3 induces extension formation and we will test if these extensions are identical to those produced by HGF. We will analyze possible effectors of PI(3,4,5)P3. Kidney diseases, including developmental defects, kidney injury and genetic conditions that form multiple cysts in the kidney, are major health problems. The kidney consists of many narrow tubes lined by cells. We are using a simple model system where kidney cells are grown in culture and produce such tubes, to study tube formation and how we can influence this to treat and prevent kidney diseases.

肾脏像许多上皮器官一样，主要由偏光层衬里的小管组成 上皮细胞。尽管我们已经了解了控制肾脏发育的基因，但我们 了解细胞将自己重新排列为小管的机制了解得多。 weuse a 在3 dimensionalCollagen生长的Madin-Darbycanine肾脏（MDCK）细胞的简化模型系统 凝胶。单个MDCK细胞植物形成由上皮细胞单层衬里的空心囊肿。当这些囊肿 被肝细胞生长因子（HGF）刺激，它们在约3天的时间内形成小管。这是一个 用于研究肾小管形成的良好模型系统，还可以提供信息多个 病理生理过程，例如从急性管状坏死中恢复 疾病，也许是干细胞的肾脏再生。我们将研究细胞的信号传导途径 回应HGF。 Erksuggest激活的动力学是由MAPK CascadeInvolving激活的动力学 B-RAF，而不是众所周知的RAF-1。我们将通过研究组件的激活来检验这一假设 B-RAF途径以及使用主导的负面因素和RNAi。我们将测试Rho的参与 在微管发生的早期阶段，家族GTPases，Rho激酶和脂质筏，尤其是形成 囊肿中细胞基底外侧表面的延伸。我们将使用延时共聚焦显微镜 观察这些分子成分对扩展形成的影响。我们将测试 磷脂酰肌醇（3,4,5）P3 [PI（3,4,5）P3]参与延伸形成。我们发现 外源性PI（3,4,5）P3诱导延伸形成，我们将测试这些扩展是否与那些相同 由HGF生产。我们将分析PI（3,4,5）P3的可能效应子。 肾脏疾病，包括发育缺陷，肾脏损伤和形成多种的遗传状况 肾脏中的囊肿是主要的健康问题。肾脏由细胞衬里的许多狭窄管组成。我们 正在使用一个简单的模型系统，其中肾细胞在培养物中生长并产生此类管，以研究 管形成，以及我们如何影响这种情况以治疗和预防肾脏疾病。