Absence of Transverse Tubules Contributes to Non-Uniform Ca2+ Wavefronts in Mouse and Human Embryonic Stem Cell-Derived Cardiomyocytes

被引:119
作者
Lieu, Deborah K. [1 ,2 ,3 ]
Liu, Jing [1 ,2 ]
Siu, Chung-Wah [1 ,4 ,5 ]
McNerney, Gregory P. [3 ]
Tse, Hung-Fat [4 ,5 ]
Abu-Khalil, Amir [1 ,2 ]
Huser, Thomas [3 ]
Li, Ronald A. [2 ,4 ,5 ,6 ]
机构
[1] Univ Calif Davis, Human Embryon Cell Consortium, Stem Cell Program, Davis, CA 95616 USA
[2] Univ Calif Davis, Dept Cell Biol & Human Anat, Davis, CA 95616 USA
[3] Univ Calif Davis, NSF, Ctr Biophoton Sci & Technol, Davis, CA 95616 USA
[4] Univ Hong Kong, Dept Med, Div Cardiol, Hong Kong, Hong Kong, Peoples R China
[5] Shriners Hosp Children N Amer, Stem Cell & Regenerat Med Programme, Heart Brain Hormone & Healthy Aging Res Ctr, Sacramento, CA USA
[6] Shriners Hosp Children N Amer, Inst Pediat Regenerat Med, Sacramento, CA USA
基金
美国国家卫生研究院;
关键词
RAT ATRIAL MYOCYTES; T-TUBULE; VENTRICULAR MYOCYTES; SKELETAL-MUSCLE; CARDIAC MYOCYTES; CALCIUM SPARKS; CONTRACTION; RELEASE; DIFFERENTIATION; BIOGENESIS;
D O I
10.1089/scd.2009.0052
中图分类号
Q813 [细胞工程];
学科分类号
摘要
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.
引用
收藏
页码:1493 / 1500
页数:8
相关论文
共 30 条
[1]   Ultrastructural comparison of developing mouse embryonic stem cell- and in vivo-derived cardiomyocytes [J].
Baharvand, Hossein ;
Piryaei, Abbas ;
Rohani, Razieh ;
Taei, Adeleh ;
Heidari, Mohammad Hassan ;
Hosseini, Ahmad .
CELL BIOLOGY INTERNATIONAL, 2006, 30 (10) :800-807
[2]   Cardiac excitation-contraction coupling [J].
Bers, DM .
NATURE, 2002, 415 (6868) :198-205
[3]   Differentiation of pluripotent embryonic stem cells into cardiomyocytes [J].
Boheler, KR ;
Czyz, J ;
Tweedie, D ;
Yang, HT ;
Anisimov, SV ;
Wobus, AM .
CIRCULATION RESEARCH, 2002, 91 (03) :189-201
[4]   Spatiotemporal characteristics of SR Ca2+ uptake and release in detubulated rat ventricular myocytes [J].
Brette, F ;
Despa, S ;
Bers, DM ;
Orchard, CH .
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY, 2005, 39 (05) :804-812
[5]   T-tubule function in mammalian cardiac myocytes [J].
Brette, F ;
Orchard, C .
CIRCULATION RESEARCH, 2003, 92 (11) :1182-1192
[6]   Resurgence of cardiac T-tubule research [J].
Brette, Fabien ;
Orchard, Clive .
PHYSIOLOGY, 2007, 22 :167-173
[7]   Amphiphysin II (SH3P9; BIN1), a member of the amphiphysin/Rvs family, is concentrated in the cortical cytomatrix of axon initial segments and nodes of Ranvier in brain and around T tubules in skeletal muscle [J].
Butler, MH ;
David, C ;
Ochoa, GC ;
Freyberg, Z ;
Daniell, L ;
Grabs, D ;
Cremona, O ;
DeCamilli, P .
JOURNAL OF CELL BIOLOGY, 1997, 137 (06) :1355-1367
[8]   Role of cholesterol in developing T-tubules: Analogous mechanisms for T-tubule and caveolae biogenesis [J].
Carozzi, AJ ;
Ikonen, E ;
Lindsay, MR ;
Parton, RG .
TRAFFIC, 2000, 1 (04) :326-341
[9]   The crystal structure of the BAR domain from human Bin1/Amphiphysin II and its implications for molecular recognition [J].
Casal, Eva ;
Federici, Luca ;
Zhang, Wei ;
Fernandez-Recio, Juan ;
Priego, Eva-Maria ;
Miguel, Ricardo Nunez ;
DuHadaway, James B. ;
Prendergast, George C. ;
Luisi, Ben F. ;
Laue, Ernest D. .
BIOCHEMISTRY, 2006, 45 (43) :12917-12928
[10]   HSP90 and eNOS partially co-localize and change cellular localization in relation to different ECM components in 2D and 3D cultures of adult rat cardiomyocytes [J].
Di Felice, Valentina ;
Cappello, Francesco ;
Montalbano, Antonella ;
Ardizzone, Nella Maria ;
De Luca, Angela ;
Macaluso, Filippo ;
Amelio, Daniela ;
Cerra, Maria Carmela ;
Zummo, Giovanni .
BIOLOGY OF THE CELL, 2007, 99 (12) :689-699