Absence of transverse tubules contributes to non-uniform Ca2+ wavefronts in mouse and human embryonic stem cell-derived cardiomyocytes

Deborah Lieu, Jing Liu, Chung Wah Siu, Gregory P. McNerney, Hung Fat Tse, Amir Abu-Khalil, Thomas R Huser, Ronald A. Li

Research output: Contribution to journalArticle

101 Citations (Scopus)

Abstract

Mouse (m) and human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are known to exhibit immature Ca2+ dynamics such as small whole-cell peak amplitude and slower kinetics relative to those of adult. In this study, we examined the maturity and efficiency of Ca2+-induced Ca2+ release in m and hESC-CMs, the presence of transverse (t) tubules and its effects on the regional Ca2+ dynamics. In m and hESC-CMs, fluorescent staining and atomic force microscopy (AFM) were used to detect the presence of t-tubules, caveolin-3, amphiphysin-2 and colocalization of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs). To avoid ambiguities, regional electrically-stimulated Ca2+ dynamics of single ESC-CMs, rather than spontaneously beating clusters, were measured using confocal microscopy. m and hESC-CMs showed absence of dyads, with neither t-tubules nor colocalization of DHPRs and RyRs. Caveolin-3 and amphiphysin-2, crucial for the biogenesis of t-tubules with robust expression in adult CMs, were also absent. Single m and hESC-CMs displayed non-uniform Ca2+ dynamics across the cell that is typical of CMs deficient of t-tubules. Local Ca2+ transients exhibited greater peak amplitude at the peripheral than at the central region for m (3.50 ± 0.42 vs. 3.05 ± 0.38) and hESC-CMs (2.96 ± 0.25 vs. 2.72 ± 0.25). Kinetically, both the rates of rise to peak amplitude and transient decay were faster for the peripheral relative to the central region. Immature m and hESC-CMs display unsynchronized Ca2+ transients due to the absence of t-tubules and gene products crucial for their biogenesis. Our results provide insights for driving the maturation of ESC-CMs.

Original languageEnglish (US)
Pages (from-to)1493-1500
Number of pages8
JournalStem Cells and Development
Volume18
Issue number10
DOIs
StatePublished - Dec 1 2009

Fingerprint

Cardiac Myocytes
Caveolin 3
L-Type Calcium Channels
Ryanodine Receptor Calcium Release Channel
Atomic Force Microscopy
Human Embryonic Stem Cells
Mouse Embryonic Stem Cells
Confocal Microscopy
Staining and Labeling
Genes

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

Cite this

Absence of transverse tubules contributes to non-uniform Ca2+ wavefronts in mouse and human embryonic stem cell-derived cardiomyocytes. / Lieu, Deborah; Liu, Jing; Siu, Chung Wah; McNerney, Gregory P.; Tse, Hung Fat; Abu-Khalil, Amir; Huser, Thomas R; Li, Ronald A.

In: Stem Cells and Development, Vol. 18, No. 10, 01.12.2009, p. 1493-1500.

Research output: Contribution to journalArticle

Lieu, Deborah ; Liu, Jing ; Siu, Chung Wah ; McNerney, Gregory P. ; Tse, Hung Fat ; Abu-Khalil, Amir ; Huser, Thomas R ; Li, Ronald A. / Absence of transverse tubules contributes to non-uniform Ca2+ wavefronts in mouse and human embryonic stem cell-derived cardiomyocytes. In: Stem Cells and Development. 2009 ; Vol. 18, No. 10. pp. 1493-1500.
@article{6c357bf8b6004d98a15c460bee62372c,
title = "Absence of transverse tubules contributes to non-uniform Ca2+ wavefronts in mouse and human embryonic stem cell-derived cardiomyocytes",
abstract = "Mouse (m) and human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are known to exhibit immature Ca2+ dynamics such as small whole-cell peak amplitude and slower kinetics relative to those of adult. In this study, we examined the maturity and efficiency of Ca2+-induced Ca2+ release in m and hESC-CMs, the presence of transverse (t) tubules and its effects on the regional Ca2+ dynamics. In m and hESC-CMs, fluorescent staining and atomic force microscopy (AFM) were used to detect the presence of t-tubules, caveolin-3, amphiphysin-2 and colocalization of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs). To avoid ambiguities, regional electrically-stimulated Ca2+ dynamics of single ESC-CMs, rather than spontaneously beating clusters, were measured using confocal microscopy. m and hESC-CMs showed absence of dyads, with neither t-tubules nor colocalization of DHPRs and RyRs. Caveolin-3 and amphiphysin-2, crucial for the biogenesis of t-tubules with robust expression in adult CMs, were also absent. Single m and hESC-CMs displayed non-uniform Ca2+ dynamics across the cell that is typical of CMs deficient of t-tubules. Local Ca2+ transients exhibited greater peak amplitude at the peripheral than at the central region for m (3.50 ± 0.42 vs. 3.05 ± 0.38) and hESC-CMs (2.96 ± 0.25 vs. 2.72 ± 0.25). Kinetically, both the rates of rise to peak amplitude and transient decay were faster for the peripheral relative to the central region. Immature m and hESC-CMs display unsynchronized Ca2+ transients due to the absence of t-tubules and gene products crucial for their biogenesis. Our results provide insights for driving the maturation of ESC-CMs.",
author = "Deborah Lieu and Jing Liu and Siu, {Chung Wah} and McNerney, {Gregory P.} and Tse, {Hung Fat} and Amir Abu-Khalil and Huser, {Thomas R} and Li, {Ronald A.}",
year = "2009",
month = "12",
day = "1",
doi = "10.1089/scd.2009.0052",
language = "English (US)",
volume = "18",
pages = "1493--1500",
journal = "Stem Cells and Development",
issn = "1547-3287",
publisher = "Mary Ann Liebert Inc.",
number = "10",

}

TY - JOUR

T1 - Absence of transverse tubules contributes to non-uniform Ca2+ wavefronts in mouse and human embryonic stem cell-derived cardiomyocytes

AU - Lieu, Deborah

AU - Liu, Jing

AU - Siu, Chung Wah

AU - McNerney, Gregory P.

AU - Tse, Hung Fat

AU - Abu-Khalil, Amir

AU - Huser, Thomas R

AU - Li, Ronald A.

PY - 2009/12/1

Y1 - 2009/12/1

N2 - Mouse (m) and human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are known to exhibit immature Ca2+ dynamics such as small whole-cell peak amplitude and slower kinetics relative to those of adult. In this study, we examined the maturity and efficiency of Ca2+-induced Ca2+ release in m and hESC-CMs, the presence of transverse (t) tubules and its effects on the regional Ca2+ dynamics. In m and hESC-CMs, fluorescent staining and atomic force microscopy (AFM) were used to detect the presence of t-tubules, caveolin-3, amphiphysin-2 and colocalization of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs). To avoid ambiguities, regional electrically-stimulated Ca2+ dynamics of single ESC-CMs, rather than spontaneously beating clusters, were measured using confocal microscopy. m and hESC-CMs showed absence of dyads, with neither t-tubules nor colocalization of DHPRs and RyRs. Caveolin-3 and amphiphysin-2, crucial for the biogenesis of t-tubules with robust expression in adult CMs, were also absent. Single m and hESC-CMs displayed non-uniform Ca2+ dynamics across the cell that is typical of CMs deficient of t-tubules. Local Ca2+ transients exhibited greater peak amplitude at the peripheral than at the central region for m (3.50 ± 0.42 vs. 3.05 ± 0.38) and hESC-CMs (2.96 ± 0.25 vs. 2.72 ± 0.25). Kinetically, both the rates of rise to peak amplitude and transient decay were faster for the peripheral relative to the central region. Immature m and hESC-CMs display unsynchronized Ca2+ transients due to the absence of t-tubules and gene products crucial for their biogenesis. Our results provide insights for driving the maturation of ESC-CMs.

AB - Mouse (m) and human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are known to exhibit immature Ca2+ dynamics such as small whole-cell peak amplitude and slower kinetics relative to those of adult. In this study, we examined the maturity and efficiency of Ca2+-induced Ca2+ release in m and hESC-CMs, the presence of transverse (t) tubules and its effects on the regional Ca2+ dynamics. In m and hESC-CMs, fluorescent staining and atomic force microscopy (AFM) were used to detect the presence of t-tubules, caveolin-3, amphiphysin-2 and colocalization of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs). To avoid ambiguities, regional electrically-stimulated Ca2+ dynamics of single ESC-CMs, rather than spontaneously beating clusters, were measured using confocal microscopy. m and hESC-CMs showed absence of dyads, with neither t-tubules nor colocalization of DHPRs and RyRs. Caveolin-3 and amphiphysin-2, crucial for the biogenesis of t-tubules with robust expression in adult CMs, were also absent. Single m and hESC-CMs displayed non-uniform Ca2+ dynamics across the cell that is typical of CMs deficient of t-tubules. Local Ca2+ transients exhibited greater peak amplitude at the peripheral than at the central region for m (3.50 ± 0.42 vs. 3.05 ± 0.38) and hESC-CMs (2.96 ± 0.25 vs. 2.72 ± 0.25). Kinetically, both the rates of rise to peak amplitude and transient decay were faster for the peripheral relative to the central region. Immature m and hESC-CMs display unsynchronized Ca2+ transients due to the absence of t-tubules and gene products crucial for their biogenesis. Our results provide insights for driving the maturation of ESC-CMs.

UR - http://www.scopus.com/inward/record.url?scp=67650022307&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650022307&partnerID=8YFLogxK

U2 - 10.1089/scd.2009.0052

DO - 10.1089/scd.2009.0052

M3 - Article

C2 - 19290776

AN - SCOPUS:67650022307

VL - 18

SP - 1493

EP - 1500

JO - Stem Cells and Development

JF - Stem Cells and Development

SN - 1547-3287

IS - 10

ER -