Na+/Ca2+ exchanger is a determinant of excitation-contraction coupling in human embryonic stem cell-derived ventricular cardiomyocytes

Ji Dong Fu, Peng Jiang, Stephanie Rushing, Jing Liu, Nipavan Chiamvimonvat, Ronald A. Li

Research output: Contribution to journalArticle

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Abstract

In adult cardiomyocytes (CMs), the Na+/Ca2+ exchanger (NCX) is a well-defined determinant of Ca2+ homeostasis. Developmentally, global NCX knockout in mice leads to abnormal myofibrillar organization, electrical defects, and early embryonic death. Little is known about the expression and function of NCX in human heart development. Self-renewable, pluripotent human embryonic stem cells (hESCs) can serve as an excellent experimental model. However, hESC-derived CMs are highly heterogeneous. A stably lentivirus-transduced hESC line (MLC2v-dsRed) was generated to express dsRed under the transcriptional control of the ventricular-restricted myosin light chain-2v (MLC2v) promoter. Electrophysiologically, dsRed+ cells differentiated from MLC2vdsRed hESCs displayed ventricular action potentials (AP), exclusively. Neither atrial nor pacemaker APs were observed. While ICa-L, If, and I Kr were robustly expressed, IKs and IK1 were absent in dsRed+ ventricular hESCCMs. Upon differentiation (7+40 to +90 days), the basal [Ca2+]i, Ca2+ transient amplitude, maximum upstroke, and decay velocities significantly increased (P<0.05). The ICa-L antagonizer nifedipine (1μM) decreased the Ca2+ transient amplitude (to ∼30%) and slowed the kinetics (by ∼2-fold), but Ca2+ transients could still be elicited even after complete ICa-L blockade, suggesting the presence of additional Ca2+ influx(es). Indeed, Ni2+-sensitive INCX could be recorded in 7+40- and +90-day dsRed+ hESC-CMs, and its densities increased from -1.2±0.6 pA/pF at -120 mV and 3.6±1.0 pA/pF at 60 mV by 6- and 2-folds, respectively. With higher [Ca2+]i, 7+90-day ventricular hESC-CMs spontaneously but irregularly fired transients upon a single stimulus under an external Na+-free condition; however, without extracellular Na +, nifedipine could completely inhibit Ca2+ transients. We conclude that INCX is functionally expressed in developing ventricular hESC-CMs and contributes to their excitation-contraction coupling.

Original languageEnglish (US)
Pages (from-to)773-782
Number of pages10
JournalStem Cells and Development
Volume19
Issue number6
DOIs
StatePublished - Jun 1 2010

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Excitation Contraction Coupling
Cardiac Myocytes
Myosin Light Chains
Nifedipine
Ventricular Myosins
Lentivirus
Human Development
Human Embryonic Stem Cells
Knockout Mice
Action Potentials
Homeostasis
Theoretical Models
fluorescent protein 583
Cell Line

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

Cite this

Na+/Ca2+ exchanger is a determinant of excitation-contraction coupling in human embryonic stem cell-derived ventricular cardiomyocytes. / Fu, Ji Dong; Jiang, Peng; Rushing, Stephanie; Liu, Jing; Chiamvimonvat, Nipavan; Li, Ronald A.

In: Stem Cells and Development, Vol. 19, No. 6, 01.06.2010, p. 773-782.

Research output: Contribution to journalArticle

Fu, Ji Dong ; Jiang, Peng ; Rushing, Stephanie ; Liu, Jing ; Chiamvimonvat, Nipavan ; Li, Ronald A. / Na+/Ca2+ exchanger is a determinant of excitation-contraction coupling in human embryonic stem cell-derived ventricular cardiomyocytes. In: Stem Cells and Development. 2010 ; Vol. 19, No. 6. pp. 773-782.
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abstract = "In adult cardiomyocytes (CMs), the Na+/Ca2+ exchanger (NCX) is a well-defined determinant of Ca2+ homeostasis. Developmentally, global NCX knockout in mice leads to abnormal myofibrillar organization, electrical defects, and early embryonic death. Little is known about the expression and function of NCX in human heart development. Self-renewable, pluripotent human embryonic stem cells (hESCs) can serve as an excellent experimental model. However, hESC-derived CMs are highly heterogeneous. A stably lentivirus-transduced hESC line (MLC2v-dsRed) was generated to express dsRed under the transcriptional control of the ventricular-restricted myosin light chain-2v (MLC2v) promoter. Electrophysiologically, dsRed+ cells differentiated from MLC2vdsRed hESCs displayed ventricular action potentials (AP), exclusively. Neither atrial nor pacemaker APs were observed. While ICa-L, If, and I Kr were robustly expressed, IKs and IK1 were absent in dsRed+ ventricular hESCCMs. Upon differentiation (7+40 to +90 days), the basal [Ca2+]i, Ca2+ transient amplitude, maximum upstroke, and decay velocities significantly increased (P<0.05). The ICa-L antagonizer nifedipine (1μM) decreased the Ca2+ transient amplitude (to ∼30{\%}) and slowed the kinetics (by ∼2-fold), but Ca2+ transients could still be elicited even after complete ICa-L blockade, suggesting the presence of additional Ca2+ influx(es). Indeed, Ni2+-sensitive INCX could be recorded in 7+40- and +90-day dsRed+ hESC-CMs, and its densities increased from -1.2±0.6 pA/pF at -120 mV and 3.6±1.0 pA/pF at 60 mV by 6- and 2-folds, respectively. With higher [Ca2+]i, 7+90-day ventricular hESC-CMs spontaneously but irregularly fired transients upon a single stimulus under an external Na+-free condition; however, without extracellular Na +, nifedipine could completely inhibit Ca2+ transients. We conclude that INCX is functionally expressed in developing ventricular hESC-CMs and contributes to their excitation-contraction coupling.",
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AU - Li, Ronald A.

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N2 - In adult cardiomyocytes (CMs), the Na+/Ca2+ exchanger (NCX) is a well-defined determinant of Ca2+ homeostasis. Developmentally, global NCX knockout in mice leads to abnormal myofibrillar organization, electrical defects, and early embryonic death. Little is known about the expression and function of NCX in human heart development. Self-renewable, pluripotent human embryonic stem cells (hESCs) can serve as an excellent experimental model. However, hESC-derived CMs are highly heterogeneous. A stably lentivirus-transduced hESC line (MLC2v-dsRed) was generated to express dsRed under the transcriptional control of the ventricular-restricted myosin light chain-2v (MLC2v) promoter. Electrophysiologically, dsRed+ cells differentiated from MLC2vdsRed hESCs displayed ventricular action potentials (AP), exclusively. Neither atrial nor pacemaker APs were observed. While ICa-L, If, and I Kr were robustly expressed, IKs and IK1 were absent in dsRed+ ventricular hESCCMs. Upon differentiation (7+40 to +90 days), the basal [Ca2+]i, Ca2+ transient amplitude, maximum upstroke, and decay velocities significantly increased (P<0.05). The ICa-L antagonizer nifedipine (1μM) decreased the Ca2+ transient amplitude (to ∼30%) and slowed the kinetics (by ∼2-fold), but Ca2+ transients could still be elicited even after complete ICa-L blockade, suggesting the presence of additional Ca2+ influx(es). Indeed, Ni2+-sensitive INCX could be recorded in 7+40- and +90-day dsRed+ hESC-CMs, and its densities increased from -1.2±0.6 pA/pF at -120 mV and 3.6±1.0 pA/pF at 60 mV by 6- and 2-folds, respectively. With higher [Ca2+]i, 7+90-day ventricular hESC-CMs spontaneously but irregularly fired transients upon a single stimulus under an external Na+-free condition; however, without extracellular Na +, nifedipine could completely inhibit Ca2+ transients. We conclude that INCX is functionally expressed in developing ventricular hESC-CMs and contributes to their excitation-contraction coupling.

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