Cell tracking using photoconvertible proteins during zebrafish development.
Cell tracking using photoconvertible proteins during zebrafish development.
复制标题
在斑马鱼发育过程中使用光转换蛋白进行细胞追踪。
DOI：
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
S. Abdelilah‐Seyfried
中科院分区：
文献类型：
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作者：
Verónica A. Lombardo;A. Sporbert;S. Abdelilah‐Seyfried
胚胎发生是一个动态的过程，最好的研究方法是使用记录体内发育变化的技术。使用基因编码的荧光蛋白已被证明是一种有价值的策略，用于阐明动态形态发生过程，因为它们发生在完整的生物体中。在过去的十年中，可光激活和光可转化荧光蛋白的发展为研究标记蛋白的离散亚群的命运打开了可能性。与光可激活蛋白不同，光可转换荧光蛋白(PCFP)在光转化之前很容易在其天然发射状态下被跟踪和成像，从而更容易通过光学检查来识别和选择区域。KAEND、KikGR、Dendra和EosFP等PCFP在紫外光或蓝光照射下可以从绿色转变为红色，这是因为His-Tyr-Gly三肽序列形成的绿色生色团可以通过光催化的β消除和随后的π-连接体系的延伸而被光转化为红色。PCFP及其单体变体分别是追踪细胞和研究蛋白质动力学的有用工具。近年来，PCFP已在不同的动物模型中表达，如斑马鱼、鸡和小鼠，用于追踪细胞命运。在这里，我们报道了一种在活的斑马鱼胚胎中对PCFP进行细胞特异性光转化的方案，并进一步追踪后期发育阶段的光转化蛋白。这种方法允许以组织特异性的方式研究斑马鱼动物模型中潜在的形态发生的细胞生物学事件。
Embryogenesis is a dynamic process that is best studied by using techniques that allow the documentation of developmental changes in vivo. The use of genetically-encoded fluorescent proteins has proven a valuable strategy for elucidating dynamic morphogenetic processes as they occur in the intact organism. During the past decade, the development of photoactivatable and photoconvertible fluorescent proteins has opened the possibility to investigate the fate of discrete subpopulations of tagged proteins. Unlike photoactivatable proteins, photoconvertible fluorescent proteins (PCFPs) are readily tracked and imaged in their native emission state prior to photoconversion, making it easier to identify and select regions by optical inspection. PCFPs, such as Kaede, KikGR, Dendra and EosFP, can be shifted from green to red upon exposure to UV or blue light due to a His-Tyr-Gly tripeptide sequence which forms a green chromophore that can be photoconverted to a red one by a light-catalyzed β-elimination and subsequent extension of a π-conjugated system. PCFPs and their monomeric variants are useful tools for tracking cells and studying protein dynamics, respectively. During recent years, PCFPs have been expressed in different animal model, such as zebrafish, chicken and mouse for cell fate tracking. Here we report a protocol for cell-specific photoconversion of PCFPs in the living zebrafish embryo and further tracking of photoconverted proteins at later developmental stages. This methodology allows studying, in a tissue-specific manner, cell biological events underlying morphogenesis in the zebrafish animal model.
影响因子：
46.2
作者：
Day RN;Davidson MW
通讯作者：
Davidson MW
影响因子：
2.7
作者：
Kulesa, Paul M.;Teddy, Jessica M.;McLennan, Rebecca
通讯作者：
McLennan, Rebecca