GLOMERULAR ENDOTHELIAL CELL BIOLOGY--DEVELOPMENT

肾小球内皮细胞生物学——发展

• 批准号: 2905825
• 负责人: BARBARA J BALLERMANN
• 金额: $ 18.15万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2905825
• 关键词: angiogenesis; apoptosis; capillary; cell adhesion; cell differentiation; cell growth regulation; cell migration; embryo /fetus tissue /cell culture; extracellular matrix proteins; fibroblast growth factor; fluorescence microscopy; growth factor; histogenesis; immunologic assay /test; integrins; laboratory rat; mitogens; molecular cloning; renal glomerulus; tissue /cell culture; transfection; transforming growth factors; transmission electron microscopy; vascular endothelial growth factors; vascular endothelium

DESCRIPTION (Adapted from the Applicant's Abstract): Glomerular capillary development involves initial ingrowth of capillary sprouts into primitive S-shaped bodies. Capillary sprouts derive either from vessels invading the developing kidney, or from vascular structures formed in situ. Initial endothelial cell proliferation and sprouting is followed by cell cycle arrest, cell flattening and fenestration. Mechanisms guiding glomerular capillary development are highly complex, involving cell-matrix and cell-cell interactions. This proposal aims to explore mechanisms whereby mediators present in the developing kidney and produced by renal epithelial cells undergoing morphogenesis influence glomerular endothelial cell capillary formation, directed tissue invasion and eventual differentiation. Insofar as normal glomerular capillary development is a prerequisite for normal glomerular function, the work is relevant to renal disease. The involvement of TGF-beta together with two endothelial cell mitogens, namely basic fibroblast growth factor (bFGF) and vascular endothelial cell growth factor (VEGF) in these processes will be explored. Their prior evidence shows that in vitro capillary formation and apoptosis are associated events and require TGF-beta mediated signals in glomerular endothelial cells. They now plan to determine whether the process of apoptosis is necessary for capillary formation, or whether it occurs only in those cells which do not have the requisite signals either from matrix and/or from the mitogens to form capillaries. They will explore whether cells protected against programmed cell death will form capillaries (instead of undergoing apoptosis) in response to TGF-beta, and to what extent protection against apoptosis is afforded by mitogens and matrix attachment. In addition to TGF-beta, they specifically plan to explore the involvement of bFGF and VEGF in the processes of capillary formation and fenestration. Extensive use will be made of glomerular capillary endothelial cells stably transfected with appropriate transdominant negative mutant receptors for appropriate growth regulators to be studied. The in vitro model systems involve a simple capillary format ion assay, collagen gel invasion assay, and co-culture with epithelial cells undergoing morphogenesis themselves. The in vivo model system involves transfection and/or transduction of post-birth developing rat kidney with the appropriate dominant negative receptor construct followed by morphological examination of glomerular capillaries at various time points during the period of ongoing glomerulogenesis.

描述（改编自申请人的摘要）：肾小球毛细血管 发育涉及毛细血管芽最初向内生长成原始的 S形体。 毛细血管芽源自侵入血管的血管 发育中的肾脏，或来自原位形成的血管结构。 最初的 内皮细胞增殖和出芽之后是细胞周期 停滞、细胞扁平化和开窗。 肾小球的引导机制 毛细血管的发育非常复杂，涉及细胞基质和 细胞与细胞的相互作用。 该提案旨在探索机制 存在于发育中的肾脏中并由肾上皮细胞产生的介质 经历形态发生的细胞影响肾小球内皮细胞 毛细血管形成、定向组织侵袭和最终分化。 正常肾小球毛细血管发育是肾小球毛细血管正常发育的先决条件 肾小球功能正常，工作与肾脏疾病有关。 TGF-β 与两种内皮细胞有丝分裂原一起参与， 即碱性成纤维细胞生长因子（bFGF）和血管内皮细胞 生长因子（VEGF）在这些过程中的作用将被探索。 他们之前的 有证据表明，体外毛细血管形成和细胞凋亡 相关事件并需要肾小球中 TGF-β 介导的信号 内皮细胞。 他们现在计划确定该过程是否 细胞凋亡是毛细血管形成所必需的，还是仅发生在 那些没有来自基质的必需信号的细胞 和/或从有丝分裂原形成毛细血管。 他们将探讨是否 免受程序性细胞死亡的细胞将形成毛细血管（而不是 细胞凋亡）对 TGF-β 的反应，以及程度如何 丝裂原和基质附着提供了针对细胞凋亡的保护。 除了TGF-beta之外，他们还特别计划探索参与 bFGF 和 VEGF 在毛细血管形成和开窗过程中的作用。 肾小球毛细血管内皮细胞将得到稳定广泛应用 用适当的转显性阴性突变受体转染 合适的生长调节剂有待研究。 体外模型系统 涉及简单的毛细管形成离子测定、胶原凝胶侵入测定、 并与本身进行形态发生的上皮细胞共培养。 体内模型系统涉及转染和/或转导 具有适当显性失活的出生后发育中的大鼠肾脏 受体构建，然后进行肾小球形态学检查 持续期间不同时间点的毛细血管 肾小球发生。