Dynamic behavior of individual cells in developing organotypic brain slices revealed by the photoconvertable protein Kaede
Dynamic behavior of individual cells in developing organotypic brain slices revealed by the photoconvertable protein Kaede
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DOI：
10.1016/j.expneurol.2006.03.022
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发表时间：
2006-08-01
影响因子：
5.3
通讯作者：
Ogawa, M.
中科院分区：
文献类型：
--
作者：
Mutoh, T.;Miyata, T.;Ogawa, M.
近年来，光学成像方法的进步促进了发育中皮层事件的可视化。特别是，将编码荧光蛋白的DNA引入到胚胎脑细胞中可以使祖细胞可视化;然后通过延时显微镜在活脑壁的情况下，可以监测转染细胞的行为。这种方法提供了大量的信息，神经元迁移的模式。然而，由于这些技术标记心室区（VZ）中的大量细胞，因此难以跟踪VZ内的单个细胞形状变化或细胞行为，其中发生神经元产生和初始迁移。在这里，我们报告了一种独特的方法，使用光转换荧光蛋白Kaede，它发出绿色荧光，并转移到发射红色荧光后，与紫外线辐射。使用这种方法，我们能够跟踪小鼠脑切片SVZ中相邻Kaede阳性细胞中一对特定子细胞的行为。梭形祖细胞分裂为两个多极子细胞。子细胞的细胞-细胞边界清晰可见，并且容易描述两个或多个细胞之间的位置和距离。Kaede的光转换特性提供了一个强大的细胞标记工具，用于识别活皮质切片内单个细胞的精确形态和迁移行为。(c)2006年爱思唯尔公司All rights reserved.
In recent years, advances in optical imaging methods have facilitated the visualization of events in the developing cortex. In particular, the introduction of DNA encoding fluorescent protein into cells of the embryonic brain allows the visualization of progenitor cells; slice preparations of the cortex then allow the monitoring of the behavior of transfected cells in the context of the living cerebral wall by time-lapse microscopy. Such approaches have provided substantial information about the patterns of neuronal migration. However, as these techniques label large numbers of cells in the ventricular zone (VZ), it is difficult to follow individual cell shape changes or cell behaviors within the VZ, where neuron production and initial migration take place. Here, we report a unique method using the photoconvertable fluorescent protein Kaede, which emits green fluorescence and shifts to emitting red fluorescence upon radiation with UV. Using this method, we were able to follow the behavior of a particular pair of daughter cells among neighboring Kaede-positive cells in the SVZ of mouse brain slices. The spindle shape progenitor divided into two multipolar-shaped daughter cells. The cell-cell borders of daughter cells were clearly visualized, and easily describe the position and distance between two or more cells. The photoconvertable property of Kaede offers a powerful cell marking tool to identify the precise morphology and migratory behaviors of individual cells within living cortical slices. (c) 2006 Elsevier Inc. All rights reserved.