Real-time analysis of ureteric bud branching morphogenesis in vitro
Real-time analysis of ureteric bud branching morphogenesis in vitro
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DOI：
10.1016/j.ydbio.2004.03.025
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发表时间：
2004-07-01
影响因子：
2.7
通讯作者：
Costantini, F
中科院分区：
文献类型：
--
作者：
Watanabe, T;Costantini, F
虽然很明显输尿管芽（UB）的正常分支形态发生对于后肾的发育至关重要，但迄今为止仅从静态图像推断出分支和生长的具体模式。在这里，我们对器官培养中小鼠肾脏早期发育过程中的 UB 分支形态发生进行了系统的延时分析。来自Hoxb7/GFP转基因胚胎的后肾原基培养3-4天，每30分钟拍摄的UB的GFP图像被组装成电影。对这些电影（可作为补充材料）的分析表明，UB 是一种高度可塑的结构，可以以各种复杂的模式进行分支，包括末端二裂、末端三裂和侧向分支。为了检查分支和伸长的动力学参数，使用 UB 的骨架表示来测量片段和分支点的数量以及每个片段的长度作为时间和分支生成的函数。这些测量结果为未来研究具有肾脏发育缺陷的突变肾脏提供了基线。为了说明如何将这些定量方法应用于肾脏发育异常的分析，我们检查了 MEK1 抑制剂 PD98059 对肾器官培养物的影响，并证实了之前的报告，即该药物对 UB 分支而不是伸长具有特定的抑制作用。 (C) 2004 Elsevier Inc. 保留所有权利。
While it is clear that the normal branching morphogenesis of the ureteric bud (UB) is critical for development of the metanephric kidney, the specific patterns of branching and growth have heretofore only been inferred from static images. Here, we present a systematic time-lapse analysis of UB branching morphogenesis during the early development of the mouse kidney in organ culture. Metanephric primordia from Hoxb7/GFP transgenic embryos were cultured for 3-4 days, and GFP images of the UB taken every 30 min were assembled into movies. Analysis of these movies (available as supplementary materials) revealed that the UB is a highly plastic structure, which can branch in a variety of complex patterns, including terminal bifid, terminal trifid, and lateral branching. To examine kinetic parameters of branching and elongation, skeletal representations of the UB were used to measure the number of segments and branch points and the length of each segment as a function of time and of branch generation. These measurements provide a baseline for future studies on mutant kidneys with defects in renal development. To illustrate how these quantitative methods can be applied to the analysis of abnormal kidney development, we examined the effects of the MEK1 inhibitor PD98059 on renal organ cultures and confirmed a previous report that the drug has a specific inhibitory effect on UB branching as opposed to elongation. (C) 2004 Elsevier Inc. All rights reserved.