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1 Division of Cardiology, Department of Medicine, Emory University , and the Atlanta Veterans Administration Medical Center, Atlanta, GA, USA; Department of Cell Biology, Emory University, Decatur, GA, USA
2 Division of Cardiology, Department of Medicine, Emory University , and the Atlanta Veterans Administration Medical Center, Atlanta, GA, USA
3 Department of Cell Biology, Emory University, Decatur, GA, USA
4 Department of Medicine, Loyola University, Maywood, IL, USA
* To whom correspondence should be addressed. E-mail: sdudley{at}emory.edu.
T-type Ca2+ channels may play a role in cardiac development. We studied the developmental regulation of the T-type currents (ICaT) in cardiomyocytes (CMs) derived from mouse embryonic stem cells. ICaT was studied in isolated CMs by whole cell patch clamp. Subsequently, CMs were identified by MLC2v driven green fluorescent protein (GFP) expression, and laser capture microdissection was used to isolate total RNA from groups of cells at various developmental time points. ICaT showed characteristics of Cav3.1, such as resistance to Ni2+ block, and a transient increase during development, correlating with measures of spontaneous electrical activity. Real-time RT-PCR showed that Cav3.1 mRNA abundance correlated (r2 = 0.81) with ICaT. The mRNA copy number was low at day 7+4 (2 copies/cell), increased significantly by day 7+10 (27/cell, p<0.01), peaked at day 7+16 (174/cell), and declined significantly at day 7+27 (25/cell). These data suggest that ICaT is developmentally regulated at the level of mRNA abundance and that this regulation parallels measures of pacemaker activity, suggesting that ICaT might play a role in the spontaneous contractions during CM development.
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