Raeppli: a whole-tissue labeling tool for live imaging of Drosophila development
Raeppli: a whole-tissue labeling tool for live imaging of Drosophila development
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DOI：
10.1242/dev.102913
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发表时间：
2014-01-15
期刊：
影响因子：
4.6
通讯作者：
Affolter, Markus
中科院分区：
文献类型：
--
作者：
Kanca, Oguz;Caussinus, Emmanuel;Affolter, Markus
观察细胞如何分裂、生长、迁移和形成发育中有机体的不同部分对于理解发育程序至关重要。在这里，我们描述了一种名为 Raeppli 的多色成像工具（以巴塞尔狂欢节上使用的彩色五彩纸屑命名）。 Raeppli 允许进行全组织标记，从而对单个器官中大多数细胞的后代进行标记，并且可以同时相对于彼此进行跟踪。我们测试了 Raeppli 在果蝇翅膀成虫盘中的使用。在幼虫阶段诱导 Raeppli 不可逆地用四种光谱可分离的明亮荧光蛋白之一以低选择偏差标记 > 90% 的细胞。为了更好地了解成虫盘的整体生长特征，我们在不同的发育阶段诱导了 Raeppli，在幼虫发育末期对多个固定盘进行了成像，并估计了它们在这些发育阶段的育儿袋原基的大小。我们还通过处于不同发育阶段的幼虫角质层对相同的翼盘进行了成像； Raeppli 标记的克隆提供了可以在多个时间点之间关联的地标。最后，我们使用 Raeppli 对翅盘前蛹外翻进行连续实时成像。
Observation of how cells divide, grow, migrate and form different parts of a developing organism is crucial for understanding developmental programs. Here, we describe a multicolor imaging tool named Raeppli (after the colorful confetti used at the carnival in Basel). Raeppli allows whole-tissue labeling such that the descendants of the majority of cells in a single organ are labeled and can be followed simultaneously relative to one another. We tested the use of Raeppli in the Drosophila melanogaster wing imaginal disc. Induction of Raeppli during larval stages irreversibly labels >90% of the cells with one of four spectrally separable, bright fluorescent proteins with low bias of selection. To understand the global growth characteristics of imaginal discs better, we induced Raeppli at various stages of development, imaged multiple fixed discs at the end of their larval development and estimated the size of their pouch primordium at those developmental stages. We also imaged the same wing disc through the larval cuticle at different stages of its development; the clones marked by Raeppli provide landmarks that can be correlated between multiple time points. Finally, we used Raeppli for continuous live imaging of prepupal eversion of the wing disc.