GLOMERULAR CAPILLARY WALL--NORMAL AND PATHOLOGIC

描述(改编自《调查员摘要》)：总体目标 这一建议的目的是研究涉及到的致病机制 糖尿病引起的胚胎后肾损害。三大潜力 目标基因将被检测为中胚层特异性基因(MEST)、H19和 基于这些基因时空表达的TIN-Ag的MEST研究 后肾间充质H19在输尿管芽上皮中的表达 和TIN-Ag在近端的反极表达，与H19在 S形状的身体表明这三个分子在早期是相互关联的 肾脏发育。实验是在五个具体目标下提出的 研究这些分子在肾脏过程中的正常和异常生物学行为 发展。I.MEST、H19和TiN-Ag的时空表达 后肾发育将通过原位杂交进行研究， 竞争性聚合酶链式反应、免疫组织化学和免疫沉淀法。二、 它们在上皮中的作用：间充质相互作用，普遍存在于 后肾发生，将通过抗体、反义、转基因和 转基因实验。三、分子蛋白质：蛋白质和RNA： 将通过重组融合来研究蛋白质的相互作用 这三个分子的不同结构的蛋白质。IV.改建 将在浓度升高的过程中对这些分子进行研究 葡萄糖在体外和高血糖状态下。五、最后， 胚胎肾脏发育过程中高血糖诱导/抑制基因的表达 将使用RDA和基因表达的系列分析进行研究 (Sage)。

DESCRIPTION (Adapted from Investigator's Abstract): The overall objective of this proposal is to study the pathogenetic mechanisms involved in diabetic induced lesions in the embryonic metanephros. Three potential target genes will be examined mesodermal specific cDNA(MEST), H19, and TIN-Ag based on the spatio-temporal expression of these genes with MEST expressed in the metanephric mesenchyme H19 in the ureteric bud epithelia and TIN-Ag in an anti-polar proximal expression opposite that of H19 in the S-shaped body suggesting the three molecules are interlinked in early nephric development. Experiments are proposed under five specific aims to study the normal and abnormal biology of these molecules during renal development. I. Spatio-temporal expression of MEST, H19 and TIN-Ag during metanephric development will be investigated by using in situ hybridization, competitive PCR, immunohistochemical and immunoprecipitation methods. II. Their role in epithelial: mesenchymal interactions, prevalent during metanephrogenesis, will be assessed by antibody, antisense, transgene and transfection experiments. III. Molecular protein: protein and RNA: protein interactions will be investigated by using recombinant fusion proteins of various constructs of these three molecules. IV. Alterations in these molecules will be investigated during elevated concentration of glucose in vitro and in hyperglycemia. V. Finally, the hyperglycemia-induced/suppressed genes during embryonic renal development will be investigated using RDA and Serial Analysis of Gene Expression (SAGE).