Chauhan Bharesh K, Reed Nathan A, Yang Ying, Cermák Lukás, Reneker Lixing, Duncan Melinda K, Cvekl Ales
The Department of Ophthalmology and Visual Sciences, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Genes Cells. 2002 Dec;7(12):1267-83. doi: 10.1046/j.1365-2443.2002.00602.x.
Pax6 is a transcription factor that is required for induction, growth, and maintenance of the lens; however, few direct target genes of Pax6 are known.
In this report, we describe the results of a cDNA microarray analysis of lens transcripts from transgenic mice over-expressing Pax6 in lens fibre cells in order to narrow the field of potential direct Pax6 target genes. This study revealed that the transcript levels were significantly altered for 508 of the 9700 genes analysed, including five genes encoding the cell adhesion molecules beta1-integrin, JAM1, L1 CAM, NCAM-140 and neogenin. Notably, comparisons between the genes differentially expressed in Pax6 heterozygous and Pax6 over-expressing lenses identified 13 common genes, including paralemmin, GDIbeta, ATF1, Hrp12 and Brg1. Immunohistochemistry and Western blotting demonstrated that Brg1 is expressed in the embryonic and neonatal (2-week-old) but not in 14-week adult lenses, and confirmed altered expression in transgenic lenses over-expressing Pax6. Furthermore, EMSA demonstrated that the BRG1 promoter contains Pax6 binding sites, further supporting the proposition that it is directly regulated by Pax6.
These results provide a list of genes with possible roles in lens biology and cataracts that are directly or indirectly regulated by Pax6.
Pax6是一种转录因子，对于晶状体的诱导、生长和维持是必需的；然而，已知的Pax6直接靶基因很少。
在本报告中，我们描述了对在晶状体纤维细胞中过表达Pax6的转基因小鼠的晶状体转录本进行cDNA微阵列分析的结果，以便缩小潜在的Pax6直接靶基因范围。这项研究表明，在所分析的9700个基因中，有508个基因的转录水平发生了显著变化，其中包括五个编码细胞粘附分子β1整合素、JAM1、L1细胞粘附分子、NCAM - 140和新基因的基因。值得注意的是，在Pax6杂合子和Pax6过表达晶状体中差异表达的基因之间的比较确定了13个共同基因，包括副勒明、GDIβ、ATF1、Hrp12和Brg1。免疫组织化学和蛋白质印迹表明，Brg1在胚胎和新生（2周龄）晶状体中表达，但在14周龄的成年晶状体中不表达，并证实了在过表达Pax6的转基因晶状体中表达发生了改变。此外，电泳迁移率变动分析表明BRG１启动子含有Pax6结合位点，进一步支持了它受Pax6直接调控的观点。
这些结果提供了一份在晶状体生物学和白内障中可能发挥作用的、直接或间接受Pax6调控的基因列表。
