Zhang WeiYan, Hawse John, Huang QingLing, Sheets Nancy, Miller Kevin M, Horwitz Joseph, Kantorow Marc
The Department of Biology, West Virginia University, Morgantown, West Virginia 26506-6057, USA.
Invest Ophthalmol Vis Sci. 2002 Jan;43(1):198-204.
To identify lens epithelial genes with altered expression levels in age-related human cataract.
Epithelia from age-related cataracts and from normal lenses were microdissected and RNA was extracted. RNAs were compared for gene expression differences by RT-PCR differential display. Transcripts exhibiting altered levels of gene expression were cloned and identified by sequencing. The expression levels of identified clones were confirmed by semiquantitative RT-PCR with three separately isolated RNA preparations. Specific primers were designed and used to examine the mRNA levels of other genes important in protein synthesis.
Numerous transcripts exhibited altered levels of gene expression. One transcript exhibiting a decreased level of expression in cataract compared with normal lenses was identified as encoding ribosomal protein L21. Three additional ribosomal proteins, L15, L13a, and L7a, also exhibited decreased expression in cataract compared with normal human lenses. By contrast, the levels of elongation factor (EF)-1alpha1 and eucaryotic initiation factor (eIF)-4E remained unchanged.
The results provide evidence that human age-related cataract is associated with decreased expression of L21 and other ribosomal proteins. The results suggest that modulation of protein synthesis and/or other functions mediated by ribosomal proteins is associated with age-related cataract.
鉴定在年龄相关性人类白内障中表达水平发生改变的晶状体上皮基因。
对年龄相关性白内障和正常晶状体的上皮进行显微切割并提取RNA。通过逆转录聚合酶链反应(RT-PCR)差异显示比较RNA的基因表达差异。对显示基因表达水平改变的转录本进行克隆并通过测序鉴定。使用三种单独分离的RNA制剂通过半定量RT-PCR确认已鉴定克隆的表达水平。设计特异性引物并用于检测蛋白质合成中其他重要基因的mRNA水平。
许多转录本显示出基因表达水平的改变。与正常晶状体相比,在白内障中表达水平降低的一种转录本被鉴定为编码核糖体蛋白L21。与正常人类晶状体相比,另外三种核糖体蛋白L15、L13a和L7a在白内障中也表现出表达降低。相比之下,延伸因子(EF)-1α1和真核起始因子(eIF)-4E的水平保持不变。
结果提供了证据表明人类年龄相关性白内障与L21和其他核糖体蛋白的表达降低有关。结果表明核糖体蛋白介导的蛋白质合成和/或其他功能的调节与年龄相关性白内障有关。