Decreased β-actin mRNA expression in hyperglycemic focal cerebral ischemia in the rat
Decreased β-actin mRNA expression in hyperglycemic focal cerebral ischemia in the rat
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DOI：
10.1016/j.neulet.2003.12.081
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发表时间：
2004-03-11
影响因子：
2.5
通讯作者：
Montgomery, J
中科院分区：
文献类型：
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作者：
Bémeur, C;Ste-Marie, L;Montgomery, J
在对脑缺血模型进行逆转录-聚合酶链反应（RT-PCR）分析时，β-肌动蛋白常被用作管家基因。在本研究中，我们测试了两个不同的控制基因用于RT-PCR实验，β-肌动蛋白和胆色素原脱氨酶（PBG-D），在大鼠模型局灶性脑缺血正常或高血糖条件下。在大鼠中使用具有永久性大脑中动脉闭塞的三血管闭塞模型。缺血后3 h，高血糖缺血大鼠的P-Actin mRNA表达低于血糖正常的缺血动物。β-肌动蛋白含量不变。至于PBG-D，其mRNA表达在各组中保持恒定。因此，我们的数据表明，在高血糖条件下局灶性脑缺血后，β-肌动蛋白是不合适的管家基因，而PBG-D是更合适的。这项研究清楚地表明了在进行RT-PCR实验时选择稳定的管家基因的重要性。(C)2003爱思唯尔爱尔兰有限公司保留所有权利。
P-Actin is often used as a housekeeping gene when performing reverse transcription-polymerase chain reaction (RT-PCR) analysis for cerebral ischemia models. In the present study, we tested two different control genes used for RT-PCR experiments, beta-actin and porphobilinogen deaminase (PBG-D), in a rat model of focal cerebral ischemia under normo- or hyperglycemic conditions. A three-vessel occlusion model with permanent middle cerebral artery occlusion was used in the rat. P-Actin mRNA expression was decreased in hyperglycemic ischemic rats compared to normoglycemic ischemic animals 3 h post-ischemia. beta-Actin protein content was unchanged. As for PBG-D, its mRNA expression remained constant throughout the groups. Our data thus show that, following focal cerebral ischemia in hyperglycemic conditions, beta-actin is an unsuitable housekeeping gene whereas PBG-D is more appropriate. This study clearly demonstrates the importance of selecting a stable housekeeping gene when performing RT-PCR experiments. (C) 2003 Elsevier Ireland Ltd. All rights reserved.