Salehi M, Hodgkins M A, Merry B J, Goyns M H
Institute for Cancer Studies, Sheffield University Medical School, United Kingdom.
Experientia. 1996 Sep 15;52(9):888-91. doi: 10.1007/BF01938876.
We have used the polymerase chain reaction (PCR)-based technique of differential display to analyse changes in gene expression during ageing of the rat brain. In this approach we have compared three young adult (6 months) with three old adult (20 months) animals. RNA preparations from the homogenised brains were subjected to reverse transcriptase (RT)-PCR using 36 different combinations of primer pairs. Any PCR product which was consistently found to be more prominent in the three young brains compared to the three old brains, and vice versa, was scored as potentially representing a gene which was differentially expressed during the ageing of this tissue. Out of a possible 2000+ PCR products we identified 44 that might represent genes that exhibit differential expression during ageing of the rat brain. An initial screen of these fragments, by Southern-blotting the PCR products and hybridising them with cDNA probes derived from either young or old brain RNA preparations, indicated that 40% of them represented genes that were differentially expressed. This approach is likely to prove invaluable for identifying cohorts of genes that show differential expression during the ageing process.
我们使用基于聚合酶链反应（PCR）的差异显示技术来分析大鼠大脑衰老过程中的基因表达变化。在这种方法中，我们将三只年轻成年（6个月）大鼠与三只老年成年（20个月）大鼠进行了比较。从匀浆大脑中提取的RNA样本，使用36种不同引物对组合进行逆转录酶（RT）-PCR。与三个老年大脑相比，在三个年轻大脑中始终更明显的任何PCR产物，反之亦然，都被视为可能代表在该组织衰老过程中差异表达的基因。在可能的2000多个PCR产物中，我们鉴定出44个可能代表在大鼠大脑衰老过程中表现出差异表达的基因。通过对PCR产物进行Southern印迹，并将它们与源自年轻或老年大脑RNA样本的cDNA探针杂交，对这些片段进行初步筛选，结果表明其中40%代表差异表达的基因。这种方法可能对于识别在衰老过程中显示差异表达的基因群体具有不可估量的价值。
