Absolute gene expression patterns of thioredoxin and glutaredoxin redox systems in mouse
Absolute gene expression patterns of thioredoxin and glutaredoxin redox systems in mouse
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DOI：
10.1074/jbc.m307866200
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发表时间：
2003-11-14
影响因子：
4.8
通讯作者：
Pueyo, C
中科院分区：
文献类型：
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作者：
Jurado, J;Prieto-Alamo, MJ;Pueyo, C
这项工作提供了小鼠中硫氧还蛋白（Trx）和谷胱甘肽（Grx）系统中所有已知组分的基因编码的第一个绝对表达模式：Trx1， Trx2, Grx1, Grx2, TrxR1, TrxR2，硫氧还蛋白/谷胱甘肽还原酶和谷胱甘肽还原酶。我们设计了一种新的分析方法，结合多重PCR和实时PCR的优点，简化了全动物实验中实际mRNA拷贝数的定量。定量报告建立了成体器官和胚胎阶段的差异，比较了mRNA衰减率，探讨了TrxR1和Grx2基因衍生的mRNA异构体的意义，并检测了百草枯诱导的超氧胁迫下的时间过程表达。总的来说，这些定量显示：i)每个转录物和所检查的小鼠器官的独特表达谱，但有一些普遍趋势，如编码硫氧还毒素的mRNA物种数量高于编码控制其氧化还原状态和活性的还原酶的mRNA物种数量；ii)在胚胎发生过程中持续表达，Trx1和TrxR1 mrna在特定时间序列上显著上调；iii) mRNA稳定性差异显著，肝衰变速率从2.8 h（硫氧还蛋白/谷胱甘肽还原酶）到大于或等于35 h （Trx1和Trx2）不等，并与mRNA稳态值直接相关；iv)产生线粒体Grx2a和67-kDa TrxR1变体的转录本的数量（相对于总同种型）的睾丸特异性差异；v) TrxR1和谷胱甘肽还原酶mrna协同上调，以器官特异性方式响应超氧化应激。进一步了解这些氧化还原系统在体内的作用应该从更集中的机制研究中获得，这些机制是在转录水平上报道的巨大差异的基础。
This work provides the first absolute expression patterns of genes coding for all known components of both thioredoxin (Trx) and glutaredoxin (Grx) systems in mouse: Trx1, Trx2, Grx1, Grx2, TrxR1, TrxR2, thioredoxin/glutathione reductase, and glutathione reductase. We devised a novel assay that, combining the advantages of multiplex and real-time PCR, streamlines the quantitation of the actual mRNA copy numbers in whole-animal experiments. Quantitations reported establish differences among adult organs and embryonic stages, compare mRNA decay rates, explore the significance of alternative mRNA isoforms derived from TrxR1 and Grx2 genes, and examine the time-course expression upon superoxide stress promoted by paraquat. Collectively, these quantitations show: i) unique expression profiles for each transcript and mouse organ examined, yet with some general trends like the higher amounts of mRNA species coding for thioredoxins than those coding for the reductases that control their redox states and activities; ii) continuous expression during embryogenesis with outstanding up-regulations of Trx1 and TrxR1 mRNAs in specific temporal sequences; iii) drastic differences in mRNA stability, liver decay rates range from 2.8 h (thioredoxin/glutathione reductase) to greater than or equal to35 h (Trx1 and Trx2), and directly correlate with mRNA steady-state values; iv) testis-specific differences in the amounts (relative to total isoforms) of transcripts yielding the mitochondrial Grx2a and 67-kDa TrxR1 variants; and v) coordinated up-regulation of TrxR1 and glutathione reductase mRNAs in response to superoxide stress in an organ-specific manner. Further insights into in vivo roles of these redox systems should be gained from more focused studies of the mechanisms underlying the vast differences reported here at the transcript level.