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, S. U. Ming-Jai, Yung Z.U. Tseng, Shoei K.Stephen Huang

Research output: Contribution to journalArticle

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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).

Original languageEnglish
Pages (from-to)214-223
Number of pages10
JournalJournal of Cardiovascular Electrophysiology
Volume15
Issue number2
DOIs
Publication statusPublished - Feb 1 2004
Externally publishedYes

Fingerprint

Myosin Light Chains
Oligonucleotide Array Sequence Analysis
Atrial Fibrillation
Electrophoresis
Protein Isoforms
Heart Atria
Proteins
Genes
Ventricular Myosins
Atrial Appendage
Messenger RNA
Swine
Complementary DNA
Clone Cells

Keywords

  • Atrial fibrillation
  • cDNA microarray
  • Mass spectrometry
  • Myosin regulatory light chain
  • Two-dimensional polyacrylamide protein electrophoresis

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

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. / Lai, Ling Ping; Lin, Jiunn Lee; Lin, Chich Sheng; Yeh, Huei Ming; Tsay, Yeou Guang; Lee, Chwen Fang; Lee, Hsiao Hui; Chang, Zee Fen; Hwang, Juey Jen; Ming-Jai, S. U.; Tseng, Yung Z.U.; Huang, Shoei K.Stephen.

In: Journal of Cardiovascular Electrophysiology, Vol. 15, No. 2, 01.02.2004, p. 214-223.

Research output: Contribution to journalArticle

Lai, Ling Ping ; Lin, Jiunn Lee ; Lin, Chich Sheng ; Yeh, Huei Ming ; Tsay, Yeou Guang ; Lee, Chwen Fang ; Lee, Hsiao Hui ; Chang, Zee Fen ; Hwang, Juey Jen ; Ming-Jai, S. U. ; Tseng, Yung Z.U. ; Huang, Shoei K.Stephen. / 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. In: Journal of Cardiovascular Electrophysiology. 2004 ; Vol. 15, No. 2. pp. 214-223.
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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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T1 - 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

AU - Lai, Ling Ping

AU - Lin, Jiunn Lee

AU - Lin, Chich Sheng

AU - Yeh, Huei Ming

AU - Tsay, Yeou Guang

AU - Lee, Chwen Fang

AU - Lee, Hsiao Hui

AU - Chang, Zee Fen

AU - Hwang, Juey Jen

AU - Ming-Jai, S. U.

AU - Tseng, Yung Z.U.

AU - Huang, Shoei K.Stephen

PY - 2004/2/1

Y1 - 2004/2/1

N2 - 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).

AB - 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).

KW - Atrial fibrillation

KW - cDNA microarray

KW - Mass spectrometry

KW - Myosin regulatory light chain

KW - Two-dimensional polyacrylamide protein electrophoresis

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