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Am J Physiol Heart Circ Physiol 285: H2770-H2779, 2003. First published August 14, 2003; doi:10.1152/ajpheart.01114.2002
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Characterization and regulation of T-type Ca2+ channels in embryonic stem cell-derived cardiomyocytes

Ying Ming Zhang,1,* Lijuan Shang,1,* Criss Hartzell,2 Michael Narlow,1 Leanne Cribbs,3 and Samuel C. Dudley, Jr.1

1Division of Cardiology, Atlanta Veterans Affairs Medical Center, Decatur 30033; 2Department of Cell Biology, Emory University, Atlanta, Georgia 30322; and 3Department of Medicine, Cardiovascular Institute, Loyola University Medical Center, Maywood, Illinois 60153

Submitted 27 December 2002 ; accepted in final form 6 August 2003

T-type Ca2+ channels may play a role in cardiac development. We studied the developmental regulation of the T-type currents (ICa,T) in cardiomyocytes (CMs) derived from mouse embryonic stem cells (ESCs). ICa,T was studied in isolated CMs by whole cell patch clamp. Subsequently, CMs were identified by the myosin light chain 2v-driven green fluorescent protein expression, and laser capture microdissection was used to isolate total RNA from groups of cells at various developmental time points. ICa,T 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 ICa,T. The mRNA copy number was low at 7+4 days (2 copies/cell), increased significantly by 7+10 days (27/cell; P < 0.01), peaked at 7+16 days (174/cell), and declined significantly at 7+27 days (25/cell). These data suggest that ICa,T is developmentally regulated at the level of mRNA abundance and that this regulation parallels measures of pacemaker activity, suggesting that ICa,T might play a role in the spontaneous contractions during CM development.

growth and development; gene expression regulation; biological pacemaker; ion channel


Address for reprint requests and other correspondence: S. C. Dudley, Jr., Div. of Cardiology, Emory Univ./Atlanta VA Medical Center, 1670 Clairmont Rd., Rm. 111B, Decatur, GA 30033 (E-mail: sdudley{at}emory.edu).




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