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用于研究斑马鱼表皮细胞类型的工具箱。

A toolbox to study epidermal cell types in zebrafish.

作者信息

Eisenhoffer George T, Slattum Gloria, Ruiz Oscar E, Otsuna Hideo, Bryan Chase D, Lopez Justin, Wagner Daniel S, Bonkowsky Joshua L, Chien Chi-Bin, Dorsky Richard I, Rosenblatt Jody

机构信息

Department of Genetics, The University of Texas MD Anderson Cancer Center, Unit 1010, 1515 Holcombe Blvd., Houston, TX 77030-4009, USA

Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, 2000 Circle of Hope Drive, Salt Lake City, UT 84112, USA.

出版信息

J Cell Sci. 2017 Jan 1;130(1):269-277. doi: 10.1242/jcs.184341. Epub 2016 May 5.

Abstract

Epithelia provide a crucial protective barrier for our organs and are also the sites where the majority of carcinomas form. Most studies on epithelia and carcinomas use cell culture or organisms where high-resolution live imaging is inaccessible without invasive techniques. Here, we introduce the developing zebrafish epidermis as an excellent in vivo model system for studying a living epithelium. We developed tools to fluorescently tag specific epithelial cell types and express genes in a mosaic fashion using five Gal4 lines identified from an enhancer trap screen. When crossed to a variety of UAS effector lines, we can now track, ablate or monitor single cells at sub-cellular resolution. Using photo-cleavable morpholino oligonucleotides that target gal4, we can also express genes in a mosaic fashion at specific times during development. Together, this system provides an excellent in vivo alternative to tissue culture cells, without the intrinsic concerns of culture conditions or transformation, and enables the investigation of distinct cell types within living epithelial tissues.

摘要

上皮组织为我们的器官提供了至关重要的保护屏障,也是大多数癌症形成的部位。大多数关于上皮组织和癌症的研究使用细胞培养或生物体,在这些模型中,如果不采用侵入性技术,就无法进行高分辨率的实时成像。在这里,我们介绍发育中的斑马鱼表皮,作为研究活体上皮组织的优秀体内模型系统。我们开发了工具,利用从增强子捕获筛选中鉴定出的5个Gal4品系,以镶嵌方式对特定上皮细胞类型进行荧光标记并表达基因。当与各种UAS效应品系杂交时,我们现在可以在亚细胞分辨率下追踪、消融或监测单个细胞。使用靶向gal4的光可裂解吗啉代寡核苷酸,我们还可以在发育过程中的特定时间以镶嵌方式表达基因。总之,该系统为组织培养细胞提供了优秀的体内替代方案,无需担心培养条件或转化问题,并且能够研究活体上皮组织内不同的细胞类型。

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