McDermott Brian M, Asai Yukako, Baucom Jessica M, Jani Shraddha D, Castellanos Yaneth, Gomez Gustavo, McClintock James M, Starr Catherine J, Hudspeth A J
Laboratory of Sensory Neuroscience and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue New York, NY 10065, USA.
Gene Expr Patterns. 2010 Feb-Mar;10(2-3):113-8. doi: 10.1016/j.gep.2010.01.001. Epub 2010 Jan 18.
The zebrafish provides a useful experimental system for investigations of aural development. To permit the controlled expression of transgenes in developing hair cells, we isolated the genomic control regions of the parvalbumin 3a (pvalb3a) and parvalbumin 3b (pvalb3b) genes. Deletion analysis and somatic-cell transgenesis restricted the cis-acting control regions for hair cells to as little as 484base pairs for pvalb3a and 650base pairs for pvalb3b. Using both meganuclease-mediated and standard methods, we produced transgenic animals that transmit transgenes through their germ lines. These fish express GFP in hair cells in the inner ear and lateral line. Two stable transgenic lines express GFP prior to hair-bundle formation, so the associated promoter constructs are suitable for manipulating gene expression during bundle development. We additionally identified a transgenic line that offers variable labeling of supporting cells.
斑马鱼为听觉发育研究提供了一个有用的实验系统。为了使转基因在发育中的毛细胞中得以可控表达,我们分离了小白蛋白3a(pvalb3a)和小白蛋白3b(pvalb3b)基因的基因组控制区域。缺失分析和体细胞转基因技术将毛细胞的顺式作用控制区域限制为:pvalb3a的低至484个碱基对,pvalb3b的为650个碱基对。我们使用大范围核酸酶介导的方法和标准方法,培育出了能通过生殖系传递转基因的转基因动物。这些鱼在内耳和侧线的毛细胞中表达绿色荧光蛋白(GFP)。两个稳定的转基因品系在毛束形成之前就表达GFP,因此相关的启动子构建体适用于在毛束发育过程中操纵基因表达。我们还鉴定出了一个能对支持细胞进行可变标记的转基因品系。