NADH-细胞色素b5还原酶在视网膜色素变性rds小鼠中的差异表达

Differential expression of NADH-cytochrome b5 reductase in the rds mouse

目的 :寻找遗传性视网膜色素变性动物模型rds小鼠发病时差异表达的基因。方法 :采用差异显示技术比较 2 5drds小鼠和正常对照小鼠的视网膜mRNA。对差异表达的基因片段进行克隆、测序和同源性比较。用基因特异引物RT -PCR观察其在不同日龄两种小鼠中的表达情况。结果 :rds小鼠与同龄正常对照小鼠相比 ,视网膜存在相当明显的基因表达差异。其中一个差异片段克隆的序列与大鼠NADH -细胞色素b5还原酶 (b5R)的cDNA序列有 91%的同源性 ,可以认为是小鼠b5R的cDNA片段。基因特异引物RT -PCR证实 12d、2 5d和 37drds小鼠视网膜中b5RmRNA水平高于同龄正常对照小鼠。结论 :可能由于某种氧化因素使b5R表达增高 ,大量消耗NADH并生成NAD+ 。

AIM: To identify the genes that were differentially expressed in the retina of rds mouse during the development of retinitis pigmentosa.METHODS: mRNA differential display method was used to compare and analyze mRNA samples prepared from the retina of rds mouse and normal mouse on postnatal day 25(P25). Differentially expressed fragments were cloned, sequenced and compared with GenBank database by BLASTN. Expression difference was further investigated by gene-specific primer RT-PCR.RESULTS: Obvious difference in gene expression occurred between rds mouse and normal mouse. One fragment, clone No.5, shared 91% homology with rat NADH-cytochrome b5 reductase (b5R) cDNA. Thus, it was identified as mouse b5R cDNA. Gene-specific RT-PCR confirmed that b5R mRNA level was increased in the retina of rds mouse compared with normal mouse on postnatal day 12, 25 and 37, respectively.CONCLUSION: Certain oxidative factors may up-regulate the expression of b5R resulting in large consumption of NADH and production of NAD +, through which apoptotic retinal cell death was enhanced.