Chen W, Hardy P, Wilce P A
Department of Biochemistry, University of Queensland, Brisbane, Australia.
Alcohol Clin Exp Res. 1997 Sep;21(6):1053-6.
The technique of polymerase chain reaction (PCR) differential display was used to detect alterations in gene expression after chronic alcohol administration. Male Wistar rats were treated with ethanol vapor for 14 days. The cDNA generated from mRNA isolated from the hippocampi of ethanol-treated and control animals was compared by PCR differential display. A differentially expressed cDNA fragment was used to screen mRNA samples by Northern analysis. The level of a mRNA was significantly elevated (x 2.5) in the hippocampus, but not the cortex of alcohol-treated rats up to 48 hr after withdrawal. Sequence analysis of the cDNA fragment revealed an almost perfect homology to rat mitochondrial NADH dehydrogenase subunit 4 mRNA. The selective induction of this mRNA in alcohol-treated rat brain areas suggests altered metabolic processes and possible dysfunction of the mitochondria. The technique of PCR differential display may prove useful in further analysis of gene expression during alcohol dependence and withdrawal.
采用聚合酶链反应（PCR）差异显示技术检测长期给予酒精后基因表达的变化。将雄性Wistar大鼠用乙醇蒸汽处理14天。通过PCR差异显示比较从乙醇处理组和对照组动物海马中分离的mRNA产生的cDNA。用一个差异表达的cDNA片段通过Northern分析筛选mRNA样本。在撤药后长达48小时，酒精处理组大鼠海马中一种mRNA的水平显著升高（增至2.5倍），但皮质中未升高。对该cDNA片段的序列分析显示，其与大鼠线粒体NADH脱氢酶亚基4 mRNA几乎完全同源。在酒精处理的大鼠脑区中该mRNA的选择性诱导提示代谢过程改变以及线粒体可能存在功能障碍。PCR差异显示技术可能在进一步分析酒精依赖和戒断期间的基因表达方面证明是有用的。
