Functional genomic study on atrial fibrillation using cDNA microarray and two-dimensional protein electrophoresis techniques and identification of the myosin regulatory light chain isoform reprogramming in atrial fibrillation
Functional genomic study on atrial fibrillation using cDNA microarray and two-dimensional protein electrophoresis techniques and identification of the myosin regulatory light chain isoform reprogramming in atrial fibrillation
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DOI：
10.1046/j.1540-8167.2004.03423.x
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发表时间：
2004-02-01
影响因子：
2.7
通讯作者：
Huang, SKS
中科院分区：
文献类型：
--
作者：
Lai, LP;Lin, JL;Huang, SKS
MLC-2 V在房颤中的上调。引言：心房组织的功能和结构变化发生在房颤（AF）的自然过程中，这些变化可能有助于进一步的AF。我们使用cDNA微阵列和二维蛋白电泳技术研究了AF组织的变化。方法和结果：我们通过快速右心耳起搏以600/min的速率建立了AF猪模型。快速心房除极6周后获得心房组织。使用含有6，035个cDNA克隆的微阵列来评估mRNA的改变。进行双向蛋白质电泳以比较蛋白质模式。在cDNA微阵列研究中，我们确定了387个基因在左心房和81个基因在右心房有显着变化。在这些基因中，心室肌球蛋白调节轻链亚型（MLC-2 V）显示出最大的变化倍数（左心房和右心房分别为9.4和7.3）。在蛋白质电泳中，在酸性区域（PI 4.5-5.0）中跨越18至20 kDa的三个蛋白质点的表达水平在AF组中特异性升高。有趣的是，通过串联质谱分析，这三个斑点被鉴定为MLC-2 V。因此，MLC-2 V的表达在mRNA和蛋白水平对应良好，都表明AF的显着增加。结论：基因芯片和双向聚丙烯酰胺蛋白电泳研究显示AF组织的特征性变化。我们证明了肌球蛋白调节轻链亚型组成的重编程，其心室亚型（MLC-2 V）显著增加。
Up-Regulation of MLC-2V in Atrial Fibrillation.Introduction: Functional and structural changes of atrial tissue occur during the natural course of atrial fibrillation (AF), and these changes may contribute to further AF. We investigated the changes in AF tissue using cDNA microarray and two-dimensional protein electrophoresis techniques.Methods and Results: We established a porcine model of AF by rapid right atrial appendage pacing at a rate of 600/min. Atrial tissue was obtained after rapid atrial depolarization for 6 weeks. Microarrays containing 6,035 cDNA clones were used to evaluate the alterations of mRNA. Two-dimensional protein electrophoresis was performed to compare protein patterns. In cDNA microarray studies, we identified 387 genes with significant change in the left atrium and 81 genes in the right atrium. Among the genes, the ventricular isoform of the myosin regulatory light chain (MLC-2V) showed the greatest fold of change (9.4 and 7.3 in the left and right atrium, respectively). In protein electrophoresis, the expression levels of three protein spots spanning from 18 to 20 kDa in the acidic region (PI 4.5-5.0) were specifically elevated in the AF group. Interestingly, through tandem mass spectrometric analysis, these three spots were identified as MLC-2V. Thus, MLC-2V expression at the mRNA and protein levels corresponded well, and both indicated a significant increase in AF.Conclusion: Both cDNA microarray and two-dimensional polyacrylamide protein electrophoresis studies revealed characteristic changes in AF tissue. We demonstrated the reprogramming of myosin regulatory light chain isoform composition, with a significant increase of its ventricular isoform (MLC-2V).