Dynamic clonal analysis based on chronic in vivo imaging allows multiscale quantification of growth in the Drosophila wing disc
Dynamic clonal analysis based on chronic in vivo imaging allows multiscale quantification of growth in the Drosophila wing disc
复制标题
DOI：
10.1242/dev.109264
复制
发表时间：
2014-06-01
期刊：
影响因子：
4.6
通讯作者：
LeGoff, Loic
中科院分区：
文献类型：
--
作者：
Heemskerk, Idse;Lecuit, Thomas;LeGoff, Loic
在形态发生过程中，组织改变形状并生长。这是如何协调的在很大程度上是未知的，部分原因是缺乏实验方法来可视化和量化增长。在这里，我们描述了一种新的实验方法来研究在体内的时间尺度的天，用于研究果蝇幼虫成虫翅盘，成虫翅的前体组织的生长。我们开发了一种协议，以图像翅盘定期在活的麻醉幼虫，以便在较长的时间内跟踪组织的生长。该方法可用于在体内以高分辨率对细胞进行成像。在中等规模下，我们跟踪了克隆内细胞数量的增加以及克隆面积和形状的变化。在扩展到组织水平的尺度上，克隆可以用作测量应变的标志，作为生长的代理。我们开发了通用的计算工具，从克隆形状和单个组织中的地标位移中提取应变图，并将多个数据集联合收割机组合成平均应变。在光盘中，我们使用这些来比较克隆（几个细胞）和更大区域尺度上的生长特性。
In the course of morphogenesis, tissues change shape and grow. How this is orchestrated is largely unknown, partly owing to the lack of experimental methods to visualize and quantify growth. Here, we describe a novel experimental approach to investigate the growth of tissues in vivo on a time-scale of days, as employed to study the Drosophila larval imaginal wing disc, the precursor of the adult wing. We developed a protocol to image wing discs at regular intervals in living anesthetized larvae so as to follow the growth of the tissue over extended periods of time. This approach can be used to image cells at high resolution in vivo. At intermediate scale, we tracked the increase in cell number within clones as well as the changes in clone area and shape. At scales extending to the tissue level, clones can be used as landmarks for measuring strain, as a proxy for growth. We developed general computational tools to extract strain maps from clonal shapes and landmark displacements in individual tissues, and to combine multiple datasets into a mean strain. In the disc, we use these to compare properties of growth at the scale of clones (a few cells) and at larger regional scales.