Lakowski Jörn, Majumder Anirban, Lauderdale James D
Department of Cellular Biology, The University of Georgia, Athens, GA 30602, USA.
Dev Biol. 2007 Jul 15;307(2):498-520. doi: 10.1016/j.ydbio.2007.04.015. Epub 2007 Apr 20.
The Pax6 gene plays several roles in retinal development, including control of cell proliferation, maintenance of the retinogenic potential of progenitor cells, and cell fate specification. Emerging evidence suggests that these different aspects of Pax6 gene function are mediated by different isoforms of the Pax6 protein; however, relatively little is known about the spatiotemporal expression of Pax6 isoforms in the vertebrate retina. Using bacterial artificial chromosome (BAC) technology, we modified a zebrafish Pax6a BAC such that we could distinguish paired-containing Pax6a transcripts from paired-less Pax6a transcripts. In the zebrafish, the spatial and temporal onset of expression of these transcripts suggests that the paired-less isoform is involved in the cell fate decision leading to the generation of amacrine cells; however, because of limitations associated with transient transgenic analysis, it was not feasible to establish whether this promoter was active in all amacrine cells or in a specific population of amacrine cells. By making mice transgenic for the zebrafish Pax6a BAC reporter transgene, we were able to show that paired-containing and paired-less Pax6a transcripts were differentially expressed in amacrine subpopulations. Our study also directly demonstrates the functional conservation of the regulatory mechanisms governing Pax6 transcription in teleosts and mammals.
Pax6基因在视网膜发育中发挥多种作用,包括控制细胞增殖、维持祖细胞的视网膜生成潜能以及细胞命运特化。新出现的证据表明,Pax6基因功能的这些不同方面是由Pax6蛋白的不同异构体介导的;然而,关于Pax6异构体在脊椎动物视网膜中的时空表达,人们了解得相对较少。利用细菌人工染色体(BAC)技术,我们对斑马鱼Pax6a BAC进行了改造,以便能够区分含有配对结构域的Pax6a转录本和不含配对结构域的Pax6a转录本。在斑马鱼中,这些转录本表达的时空起始表明,不含配对结构域的异构体参与了导致无长突细胞产生的细胞命运决定;然而,由于与瞬时转基因分析相关的局限性,确定该启动子是否在所有无长突细胞或特定的无长突细胞群体中活跃是不可行的。通过制作携带斑马鱼Pax6a BAC报告转基因的转基因小鼠,我们能够表明含有配对结构域和不含配对结构域的Pax6a转录本在无长突细胞亚群中差异表达。我们的研究还直接证明了硬骨鱼和哺乳动物中调控Pax6转录的机制在功能上具有保守性。