Abstract
Aim:
To investigate the developmental regulation of intracellular Ca2+ transients, an essential event in excitation-contraction coupling, during cardiomyocyte differentiation.
Methods:
Using the embryonic stem (ES) cell in vitro differentiation system and pharmacological intervention, we investigated the molecular and functional regulation of Ca2+ handling proteins on the Ca2+ transients at early, intermediate and later differentiation stages of ES cell-derived cardiomyocytes (ESCM).
Results:
Nifedipine, a selective antagonist of L-type Ca2+ channels, totally blocked Ca2+ transients even in the condition of field-electric stimulation in ESCM at three differentiation stages. The Ca2+ transients of ESCM were also inhibited by both ryanodine [an inhibitor of ryanodine receptors (RyRs)] and 2-aminoethoxydipheylborate [2-APB, an inhibitor of inositol-1,4,5-trisphosphate receptors (IP3Rs)]. The inhibitory effect of ryanodine increased with the time of differentiation, while the effect of 2-APB decreased with the differentiation. Thapsigargin, an inhibitor of SR Ca2+-pump ATPase, inhibited Ca2+ transients equally at three differentiation stages that matched the expression profile. Na+ free solution, which inhibits Na+-Ca2+ exchanger (NCX) to extrude Ca2+ from cytosol, did not affect the amplitude of Ca2+ transients of ESCM until the latter differentiation stage, but it significantly enhanced the basal Ca2+concentration.
Conclusion:
The Ca2+ transients in ESCM depend on both the sarcolemmal Ca2+ entry via L-type Ca2+ channels and the SR Ca2+ release from RyRs and IP3Rs even at the early differentiation stage; but NCX seems not to regulate the peak of Ca2+ transients until the latter differentiation stage.
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References
Clapham DE . Calcium signaling. Cell 1995; 80: 259–68.
Webb SE, Miller AL . Calcium signaling during embryonic development. Nat Rev Mol Cell Biol 2003; 4: 539–51.
Puceat M, Jaconi M . Ca(2+) signaling in cardiogenesis. Cell Calcium 2005; 38: 383–9.
Evenas J, Malmendal A, Forsen S . Calcium. Curr Opin Chem Biol 1998; 2: 293–302.
Barry WH, Bridge JH . Intracellular calcium homeostasis in cardiac myocytes. Circulation 1993; 87: 1806–15.
Bers DM . Cardiac excitation-contraction coupling. Nature 2002; 415: 198–205.
Brand T . Heart development: molecular insights into cardiac specification and early morphogenesis. Dev Biol 2003; 258: 1–19.
Liu W, Yasui K, Opthof T, Ishiki R, Lee JK, Kamiya K, et al. Developmental changes of Ca(2+) handling in mouse ventricular cells from early embryo to adulthood. Life Sci 2002; 71: 1279–92.
Reed TD, Babu GJ, Ji Y, Zilberman A, Ver Heyen M, Wuytack F, et al. The expression of SR calcium transport ATPase and the Na (+)/Ca(2+)exchanger are antithetically regulated during mouse cardiac development and in hypo/hyperthyroidism. J Mol Cell Cardiol 2000; 32: 453–64.
Hescheler J, Fleischmann BK, Lentini S, Maltsev VA, Rohwedel J, Wobus AM, et al. Embryonic stem cells: a model to study structural and functional properties in cardiomyogenesis. Cardiovasc Res 1997; 36: 149–62.
Westfall MV, Pasyk KA, Yule DI, Samuelson LC, Metzger JM . Ultrastructure and cell-cell coupling of cardiac myocytes differentiating in embryonic stem cell cultures. Cell Motil Cytoskeleton 1997; 36: 43–54.
Boheler KR, Czyz J, Tweedie D, Yang HT, Anisimov SV, Wobus AM . Differentiation of pluripotent embryonic stem cells into cardiomyocytes. Circ Res 2002; 91: 189–201.
Banach K, Halbach MD, Hu P, Hescheler J, Egert U . Development of electrical activity in cardiac myocyte aggregates derived from mouse embryonic stem cells. Am J Physiol Heart Circ Physiol 2003; 284: H2114–23.
Doevendans PA, Kubalak SW, An RH, Becker DK, Chien KR, Kass RS . Differentiation of cardiomyocytes in floating embryoid bodies is comparable to fetal cardiomyocytes. J Mol Cell Cardiol 2000; 32: 839–51.
Fijnvandraat AC, van Ginneken AC, Schumacher CA, Boheler KR, Lekanne Deprez RH, Christoffels VM, et al. Cardiomyocytes purified from differentiated embryonic stem cells exhibit characteristics of early chamber myocardium. J Mol Cell Cardiol 2003; 35: 1461–72.
Wobus AM, Guan K, Yang HT, Boheler KR . Embryonic stem cells as a model to study cardiac, skeletal muscle, and vascular smooth muscle cell differentiation. Methods Mol Biol 2002; 185: 127–56.
Fu JD, Li J, Tweedie D, Yu HM, Chen L, Wang R, et al. Crucial role of the sarcoplasmic reticulum in the developmental regulation of Ca2+ transients and contraction in cardiomyocytes derived from embryonic stem cells. FASEB J 2006; 20: 181–3.
Yang HT, Tweedie D, Wang S, Guia A, Vinogradova T, Bogdanov K, et al. The ryanodine receptor modulates the spontaneous beating rate of cardiomyocytes during development. Proc Natl Acad Sci USA 2002; 99: 9225–30.
Grynkiewicz G, Poenie M, Tsien RY . A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440–50.
Tibbits GF, Xu L, Sedarat F . Ontogeny of excitation-contraction coupling in the mammalian heart. Comp Biochem Physiol A Mol Integr Physiol 2002; 132: 691–8.
Liu W, Yasui K, Arai A, Kamiya K, Cheng J, Kodama I, et al. beta-adrenergic modulation of L-type Ca2+-channel currents in early-stage embryonic mouse heart. Am J Physiol 1999; 276: H608–13.
Maltsev VA, Ji GJ, Wobus AM, Fleischmann BK, Hescheler J . Establishment of beta-adrenergic modulation of L-type Ca2+ current in the early stages of cardiomyocyte development. Circ Res 1999; 84: 136–45.
Maruyama T, Kanaji T, Nakade S, Kanno T, Mikoshiba K . 2APB, 2-aminoethoxydiphenyl borate, a membrane-penetrable modulator of Ins(1,4,5)P3-induced Ca2+ release. J Biochem (Tokyo) 1997; 122: 498–505.
Bootman MD, Collins TJ, Mackenzie L, Roderick HL, Berridge MJ, Peppiatt CM . 2-aminoethoxydiphenyl borate (2-APB) is a reliable blocker of store-operated Ca2+ entry but an inconsistent inhibitor of InsP3-induced Ca2+ release. FASEB J 2002; 16: 1145–50.
Tohse N, Meszaros J, Sperelakis N . Developmental changes in long-opening behavior of L-type Ca2+ channels in embryonic chick heart cells. Circ Res 1992; 71: 376–84.
Davies MP, An RH, Doevendans P, Kubalak S, Chien KR, Kass RS . Developmental changes in ionic channel activity in the embryonic murine heart. Circ Res 1996; 78: 15–25.
Masuda H, Sumii K, Sperelakis N . Long openings of calcium channels in fetal rat ventricular cardiomyocytes. Pflugers Arch 1995; 429: 595–7.
Nakanishi T, Seguchi M, Takao A . Development of the myocardial contractile system. Experientia 1988; 44: 936–44.
Mahony L, Jones LR . Developmental changes in cardiac sarco-plasmic reticulum in sheep. J Biol Chem 1986; 261: 15257–65.
Pegg W, Michalak M . Differentiation of sarcoplasmic reticulum during cardiac myogenesis. Am J Physiol 1987; 252: H22–31.
Olivetti G, Anversa P, Loud AV . Morphometric study of early postnatal development in the left and right ventricular myocardium of the rat. II. Tissue composition, capillary growth, and sarcoplasmic alterations. Circ Res 1980; 46: 503–12.
Takeshima H, Komazaki S, Hirose K, Nishi M, Noda T, Iino M . Embryonic lethality and abnormal cardiac myocytes in mice lacking ryanodine receptor type 2. EMBO J 1998; 17: 3309–16.
Seki S, Nagashima M, Yamada Y, Tsutsuura M, Kobayashi T, Namiki A, et al. Fetal and postnatal development of Ca2+ transients and Ca2+ sparks in rat cardiomyocytes. Cardiovasc Res 2003; 58: 535–48.
Marks AR . Intracellular calcium-release channels: regulators of cell life and death. Am J Physiol 1997; 272: H597–605.
Rosemblit N, Moschella MC, Ondriasa E, Gutstein DE, Ondrias K, Marks AR . Intracellular calcium release channel expression during embryogenesis. Dev Biol 1999; 206: 163–77.
Saeki T, Shen JB, Pappano AJ . Inositol-1,4,5-trisphosphate increases contractions but not L-type calcium current in guinea pig ventricular myocytes. Cardiovasc Res 1999; 41: 620–8.
Mery A, Aimond F, Menard C, Mikoshiba K, Michalak M, Puceat M . Initiation of embryonic cardiac pacemaker activity by inositol 1,4,5-trisphosphate-dependent calcium signaling. Mol Biol Cell 2005; 16: 2414–23.
Koban MU, Moorman AF, Holtz J, Yacoub MH, Boheler KR . Expressional analysis of the cardiac Na-Ca exchanger in rat development and senescence. Cardiovasc Res 1998; 37: 405–23.
Qu Y, Ghatpande A, el Sherif N, Boutjdir M . Gene expression of Na+/Ca2+ exchanger during development in human heart. Cardiovasc Res 2000; 45: 866–73.
Ghatpande S, Goswami S, Mascareno E, Siddiqui MA . Signal transduction and transcriptional adaptation in embryonic heart development and during myocardial hypertrophy. Mol Cell Biochem 1999; 196: 93–7.
Marks AR . Ryanodine receptors/calcium release channels in heart failure and sudden cardiac death. J Mol Cell Cardiol 2001; 33: 615–24.
Soonpaa MH, Koh GY, Klug MG, Field LJ . Formation of nascent intercalated disks between grafted fetal cardiomyocytes and host myocardium. Science 1994; 264: 98–101.
Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, et al. Bone marrow cells regenerate infarcted myocardium. Nature 2001; 410: 701–5.
Min JY, Yang Y, Converso KL, Liu L, Huang Q, Morgan JP, et al. Transplantation of embryonic stem cells improves cardiac function in postinfarcted rats. J Appl Physiol 2002; 92: 288–96.
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Project supported in part by grants of the National Natural Science Foundation of China (No 30270656), the National Natural Science Foundation of China and The Hong Kong Research Grants Council(NSFC-RGC, 30518003) and Programs (03DJ14019) from Science and Technology Committee of Shanghai Municipality.
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Fu, Jd., Yu, Hm., Wang, R. et al. Developmental regulation of intracellular calcium transients during cardiomyocyte differentiation of mouse embryonic stem cells. Acta Pharmacol Sin 27, 901–910 (2006). https://doi.org/10.1111/j.1745-7254.2006.00380.x
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DOI: https://doi.org/10.1111/j.1745-7254.2006.00380.x
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