Shi S, Bekhor I
Laboratory for Lens Molecular Biology, Doheny Eye Institute, Los Angeles, CA 90033.
Exp Eye Res. 1994 Aug;59(2):171-7. doi: 10.1006/exer.1994.1095.
Oxidative mechanisms are thought to play a major role in several biological phenomena, including cataract formation. In the following studies we determined the relative levels of expression of the genes for the mRNAs for glutathione peroxidase (GPx), glutathione reductase (GR), CuZn-superoxide dismutase (CuZn-SOD) and catalase, in both the rat lens and liver. Northern blot hybridization methods were used to determine the mRNA size. The RNase protection method was used to determine levels of expression for these mRNAs plus levels of expression for alpha A-crystallin and gamma-crystallin mRNAs in the lens, and gamma-actin mRNAs in both the lens and the liver; using [32P]-labeled specific cRNA probes transcribed from the various cDNA clones for the mRNAs being studied. The data was normalized relative to the level of expression of alpha A-crystallin and gamma-actin mRNAs in the lens, and to gamma-actin mRNA in the liver. We find the levels of the mRNAs in the lens fall in the following descending order: GPx > GR > CuZn-SOD > catalase, in the same order as has been reported for the activities of the enzymes in the lens. In the liver, levels of these mRNAs were as follows: GPx > CuZn-SOD > GR > catalase. In the liver, CuZn-SOD mRNA was expressed at about four times the level found in the lens, GPx at three times, catalase at three times and GR at about the same level.(ABSTRACT TRUNCATED AT 250 WORDS)
氧化机制被认为在包括白内障形成在内的多种生物学现象中起主要作用。在接下来的研究中,我们测定了大鼠晶状体和肝脏中谷胱甘肽过氧化物酶(GPx)、谷胱甘肽还原酶(GR)、铜锌超氧化物歧化酶(CuZn-SOD)和过氧化氢酶基因的mRNA相对表达水平。采用Northern印迹杂交法测定mRNA大小。采用核糖核酸酶保护法测定这些mRNA的表达水平,以及晶状体中αA-晶状体蛋白和γ-晶状体蛋白mRNA的表达水平,以及晶状体和肝脏中γ-肌动蛋白mRNA的表达水平;使用从用于研究的各种mRNA的cDNA克隆转录的[32P]标记的特异性cRNA探针。数据相对于晶状体中αA-晶状体蛋白和γ-肌动蛋白mRNA的表达水平以及肝脏中γ-肌动蛋白mRNA的水平进行归一化。我们发现晶状体中mRNA的水平按以下降序排列:GPx>GR>CuZn-SOD>过氧化氢酶,与晶状体中酶活性的报道顺序相同。在肝脏中,这些mRNA的水平如下:GPx>CuZn-SOD>GR>过氧化氢酶。在肝脏中,CuZn-SOD mRNA的表达水平约为晶状体中的四倍,GPx为三倍,过氧化氢酶为三倍,GR的表达水平大致相同。(摘要截短至250字)