Details

Autor(en) / Beteiligte

• Fang, Xiefan
• Poulsen, Ryan R.
• Wang‐Hu, John
• Shi, Olivia
• Calvo, Nicholas S.
• Simmons, Chelsey S.
• Rivkees, Scott A.
• Wendler, Christopher C.

Titel

Knockdown of DNA methyltransferase 3a alters gene expression and inhibits function of embryonic cardiomyocytes

Ist Teil von

• The FASEB journal, 2016-09, Vol.30 (9), p.3238-3255

Ort / Verlag

United States: Federation of American Societies for Experimental Biology

Erscheinungsjahr

2016

Quelle

Wiley-Blackwell Journals

Beschreibungen/Notizen

• ABSTRACT We previously found that in utero caffeine exposure causes down‐regulation of DNA methyltransferases (DNMTs) in embryonic heart and results in impaired cardiac function in adulthood. To assess the role of DNMTs in these events, we investigated the effects of reduced DNMT expression on embryonic cardiomyocytes. siRNAs were used to knock down individual DNMT expression in primary cultures of mouse embryonic cardiomyocytes. Immunofluorescence staining was conducted to evaluate cell morphology. A video‐based imaging assay and multi‐electrode array were used to assess cardiomyocyte contractility and electrophysiology, respectively. RNA‐Seq and multiplex bisulfite sequencing were performed to examine gene expression and promoter methylation, respectively. At 72 h after transfection, reduced DNMT3a expression, but not DNMT1 or −3b, disrupted sarcomere assembly and decreased beating frequency, contractile movement, amplitude of field action potential, and cytosolic calcium signaling of cardiomyocytes. RNA‐Seq analysis revealed that the DNMT3a‐deficient cells had deactivated gene networks involved in calcium, endothelin‐1, renin‐angiotensin, and cardiac b‐adrenergic receptor signaling, which were not inhibited by DNMT3b siRNA. Moreover, decreased methylation levels were found in the promoters of Myh7, Myh7b, Tnni3,and Tnnt2, consistent with the up‐regulation of these genes by DNMT3a siRNA. These data show that DNMT3a plays an important role in regulating embryonic cardiomyocyte gene expression, morphology and function.—Fang, X., Poulsen, R. R., Wang‐Hu, J., Shi,O., Calvo, N. S., Simmons, C. S., Rivkees, S. A., Wendler, C. C. Knockdown of DNA methyltransferase 3a alters gene expression and inhibits function of embryonic cardiomyocytes. FASEB J. 30, 3238–3255 (2016). www.fasebj.org