Chauhan Bharesh K, Reed Nathan A, Zhang Weijia, Duncan Melinda K, Kilimann Manfred W, Cvekl Ales
Department of Ophthalmology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
J Biol Chem. 2002 Mar 29;277(13):11539-48. doi: 10.1074/jbc.M110531200. Epub 2002 Jan 14.
Pax6 is a transcription factor that regulates the development of the visual, olfactory, and central nervous systems, pituitary, and pancreas. Pax6 is required for induction, growth, and maintenance of the lens; however, few direct Pax6 target genes are known. This study was designed to identify batteries of differentially expressed genes in three related systems: 8-week old Pax6 heterozygous lenses, 8-week old Pax6 heterozygous eyes, and transgenic lenses overexpressing PAX6(5a), using high throughput cDNA microarrays containing about 9700 genes. Initially, we obtained almost 400 differentially expressed genes in lenses from mice heterozygous for a Pax6 deletion, suggesting that Pax6 haploinsufficiency causes global changes in the lens transcriptome. Comparisons between the three sets of analyses revealed that paralemmin, molybdopterin synthase sulfurylase, Tel6 oncogene (ETV6), a cleavage-specific factor (Cpsf1) and tangerin A were abnormally expressed in all three experimental models. Semiquantitative reverse transcription (RT)-PCR analysis confirmed that all five of these genes were differentially expressed in Pax-6 heterozygous and Pax6(5a) transgenic lenses. Western blotting and immunohistochemistry demonstrated that paralemmin is found at high levels in the adult lens and confirmed its down-regulation in the Pax6(5a)-transgenic lenses. Collectively, our data provide insights into the genetic programs regulated by Pax6 in the lens.
Pax6是一种转录因子，可调节视觉、嗅觉和中枢神经系统、垂体及胰腺的发育。晶状体的诱导、生长和维持需要Pax6；然而，已知的直接受Pax6调控的靶基因很少。本研究旨在利用包含约9700个基因的高通量cDNA微阵列，鉴定三个相关系统中差异表达的基因群：8周龄的Pax6杂合晶状体、8周龄的Pax6杂合眼以及过表达PAX6(5a)的转基因晶状体。最初，我们在Pax6缺失杂合小鼠的晶状体中获得了近400个差异表达基因，这表明Pax6单倍体不足会导致晶状体转录组的整体变化。三组分析之间的比较显示，在所有三个实验模型中，副勒明、钼蝶呤合酶硫化酶、Tel6致癌基因（ETV6）、一种切割特异性因子（Cpsf1）和柑橘素A均异常表达。半定量逆转录（RT）-PCR分析证实，这五个基因在Pax-6杂合和Pax6(5a)转基因晶状体中均有差异表达。蛋白质免疫印迹和免疫组织化学表明，副勒明在成年晶状体中高水平表达，并证实其在Pax6(5a)转基因晶状体中表达下调。总体而言，我们的数据为Pax6在晶状体中调控的遗传程序提供了见解。
