基于连续切片的数字化三维重建小鼠发育肾单位

项目名称： 基于连续切片的数字化三维重建小鼠发育肾单位

项目编号： No.30871313

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 翟效月

作者单位： 中国医科大学

项目金额： 35万元

中文摘要： 文献中，小鼠发育肾组织形态演变及细胞生物学信息不详甚至缺如。利用前期建立起来的生物医学标本制备并结合计算机图像分析及数据处理的技术，本研究建立了肾发育中多个时间点的显微序列图像库，并对胎龄17天和生后7天小鼠肾的各种管状微细结构进行了三维追踪，在此过程中，建立了发育不同阶段的肾单位及血管走行坐标数据库，进而将追踪小管和血管的走行及毗邻关系可视化，以便系统地描述集合管系的走行、分支和与肾单位的连接方式。在建立图像库和坐标数据库的基础上，利用三维重建的切片，对发生发育肾的细胞增殖及凋亡以及多种上皮转运体的定位进行了研究。其中，获得的重要发现是：BCL-2/BAX在发育肾不同小管节段的表达存在差异，始终共表达在较成熟的近端小管，尤其近曲小管，提示近端小管对凋亡诱导的敏感，对肾脏缺血、缺氧、中毒等急性肾损伤的病理机制提供了细胞生物学依据。然而，BCL-2/BAX是否与肾小管的生长发育和凋亡有关还尚需进一步研究。此外，膜转运体的发现提示，蛋白和水重吸收的转运体出现较早，尿素转运体较晚，提示尿液浓缩功能在小鼠形成的较晚。此项研究的数据还在整理和分析中，可望今年发表2-3篇SCI论文。

中文关键词： 发生发育肾；凋亡相关蛋白；三维重建；上皮转运体；免疫细胞化学

英文摘要： Abstract: In literatures, the morphogenesis and cell biological information of developing kidney are not concrete and even incomplete. Using previously established techniques combining biomedical sample preparation and image digitalization and computer programs, the present study set up 2 databases: one for series microscopic images derived from developing kidneys on multiple time points，the other recording automatically coordinates of the tubule course traced at embryonic day 17 and postnatal day 7 kidneys. Thereafter, the spatial arrangement of the tubules could be visualized for analyzing the course and branching and the connection with nephrons of collecting duct system. Based on the databases, cellular activities, e.g. apoptosis and proliferation and epithelial transporters, were localized on the sections that were used for 3D reconstruction. Among the results, that the BCL-2/BAX were consistently expressed along mature proximal tubules and differentially expressed in other tubule segments was emphasized, suggesting a differential reaction and pathologic process of the tubules under conditions, such as ischemia, hypoxia, toxicity, and high glucose level. Whether or not the BCL-2/BAX play roles in apoptosis or cellular differentiation during kidney development is need further explored. In addition, the results from epithelial transporter studies implied that the reabsorption of proteins and waters facilitated by Megalin and Cubilin and aquaporins may occurred as early as embryonic day 14 whereas the urea transporter was detected not before postnatal day 7, suggesting a poor urine concentration capacity during early and middle stage of kidney development.So far, the experimental data are being collected, documented, and analyzed, hoping 2 or 3 publications come out within the year.

英文关键词： developing kidney; apoptosis related proteins; 3D reconstruction; epithelial transporters; immunohistocytochemistry