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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.

Ling-Ping Lai·Jiunn-Lee Lin·Chich-Sheng Lin·Huei-Ming Yeh·Yeou-Guang Tsay·Chwen-Fang Lee·Hsiao-Hui Lee·Zee-Fen Chang·Juey-Jen Hwang·Ming-Jai Su·Yung-Zu Tseng·Shoei K Stephen Huang
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Abstract

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).

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AnimalsAtrial FibrillationBase SequenceCardiac Pacing, ArtificialDisease Models, AnimalElectrophoresis, Gel, Two-DimensionalGene Expression ProfilingGene Expression RegulationGenomic LibraryHeart AtriaHeart Conduction SystemHeart VentriclesMass SpectrometryModels, CardiovascularMolecular Sequence DataMyosin Light ChainsOligonucleotide Array Sequence AnalysisProtein IsoformsReverse Transcriptase Polymerase Chain ReactionSwineComparative StudyJournal ArticleResearch Support, Non-U.S. Gov't

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Ling-Ping Lai, Jiunn-Lee Lin, Chich-Sheng Lin, Huei-Ming Yeh, Yeou-Guang Tsay, Chwen-Fang Lee, Hsiao-Hui Lee, Zee-Fen Chang, Juey-Jen Hwang, Ming-Jai Su, Yung-Zu Tseng, Shoei K Stephen Huang. 2004. 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.. https://doi.org/10.1046/j.1540-8167.2004.03423.x

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