Pfaffl Michael W, Gerstmayer B, Bosio A, Windisch Wilhelm
Institute of Physiology, Department of Animal Sciences, Centre of Life and Food Sciences, Technical University of Munich, 85354 Freising, Germany.
J Nutr Biochem. 2003 Dec;14(12):691-702. doi: 10.1016/j.jnutbio.2003.08.007.
In the study presented here, the effect of zinc deficiency on mRNA expression levels in liver and jejunum of adult rats was analyzed. Feed intake was restricted to 8 g/day. The semi-synthetic diet was fortified with pure phytate and contained either 2 microg Zn/g (Zn deficiency, n = 6) or 58 microg Zn/g (control, n = 7). After 29 days of Zn depletion feeding, entire jejunum and liver were retrieved and total RNA was extracted. Tissue specific expression pattern were screened and quantified by microarray analysis and verified individually via real-time RT-PCR. A relative quantification was performed with the newly developed Relative Expression Software Tool Copyright on numerous candidate genes which showed a differential expression. This study provides the first comparative view of gene expression regulation and fully quantitative expression analysis of 35 candidate genes in a non-growing Zn deficient adult rat model. The expression results indicate the existence of individual expression pattern in liver and jejunum and their tissue specific regulation under Zn deficiency. In addition, in jejunum a number of B-cell related genes could be demonstrated to be suppressed at Zn deficiency. In liver, metallothionein subtype 1 and 2 (MT-1 and MT-2) genes could be shown to be dramatically repressed and therefore represent putative markers for Zn deficiency. Expression results imply that some genes are expressed constitutively, whereas others are highly regulated in tissues responsible for Zn homeostasis.
在本研究中,分析了锌缺乏对成年大鼠肝脏和空肠中mRNA表达水平的影响。采食量限制为每天8克。半合成饲料添加了纯植酸盐,含有2微克锌/克(锌缺乏组,n = 6)或58微克锌/克(对照组,n = 7)。在进行29天的低锌喂养后,取出整个空肠和肝脏并提取总RNA。通过微阵列分析筛选并定量组织特异性表达模式,并通过实时RT-PCR进行个体验证。使用新开发的相对表达软件工具版权对众多显示差异表达的候选基因进行相对定量。本研究首次比较了非生长性锌缺乏成年大鼠模型中35个候选基因的基因表达调控和完全定量表达分析。表达结果表明肝脏和空肠中存在个体表达模式以及锌缺乏情况下它们的组织特异性调控。此外,在空肠中,许多B细胞相关基因在锌缺乏时可被证明受到抑制。在肝脏中,金属硫蛋白亚型1和2(MT-1和MT-2)基因可被证明受到显著抑制,因此代表锌缺乏的推定标志物。表达结果表明,一些基因是组成性表达,而其他基因在负责锌稳态的组织中受到高度调控。