Turn off MathJax
Article Contents
Fan Xu, Jing-Jing Li, Eric Yang, Yi Zhang, Wenjun Xie. Assaying sarcoplasmic reticulum Ca2+-leak in mouse atrial myocytes. Biophysics Reports. doi: 10.52601/bpr.2023.230044
Citation: Fan Xu, Jing-Jing Li, Eric Yang, Yi Zhang, Wenjun Xie. Assaying sarcoplasmic reticulum Ca2+-leak in mouse atrial myocytes. Biophysics Reports. doi: 10.52601/bpr.2023.230044

Assaying sarcoplasmic reticulum Ca2+-leak in mouse atrial myocytes

doi: 10.52601/bpr.2023.230044
More Information
  • Corresponding author: xiewenjun@xjtu.edu.cn (W. Xie)
  • Received Date: 06 December 2023
  • Accepted Date: 26 December 2023
  • Available Online: 26 February 2024
  • More and more studies have suggested an essential role of sarcoplasmic reticulum (SR) Ca2+ leak of atrial myocytes in atrial diseases such as atrial fibrillation (AF). The increasing interest in atrial Ca2+ signaling makes it necessary to develop a more accurate approach for Ca2+ measurement in atrial myocytes due to obvious differences between atrial and ventricular Ca2+ handling. In the present study, we proposed a new approach for quantifying total SR Ca2+ leak in atrial myocytes with confocal line-scan Ca2+ images. With a very precious approximation of the histogram of normalized line-scan Ca2+ images by using a modified Gaussian distribution, we separated the signal pixel components from noisy pixels and extracted two new dimensionless parameters, Fsignals and Rsignals, to reflect the summation of signal pixels and their release components, respectively. In the presence of tetracaine blocking SR Ca2+ leak, the two parameters were very close to 0, and in atrial myocytes under normal conditions, the two parameters are well positive correlative with Ca2+ spark frequency and total signal mass, the two classic readouts for SR Ca2+ leak. Consistent with Ca2+ Spark readouts, the two parameters quantified a significant increase of SR Ca2+ leak in atrial myocytes from mice harboring a leaky type 2 ryanodine receptor mutation (RyR2-R2474S+/−) compared to the WT group. Collectively, this study proposed a simple and effective approach to quantify SR Ca2+ leak in atrial myocytes, which may benefit research on calcium signaling in atrial physiology and diseases.

  • Fan Xu, Jing-Jing Li, Eric Yang, Yi Zhang and Wenjun Xie declare that they have no conflict of interest.
    All institutional and national guidelines for the care and use of laboratory animals were followed.

  • loading
  • Bers DM (2002) Cardiac excitation-contraction coupling. Nature 415(6868): 198−205 doi: 10.1038/415198a
    Bootman MD, Smyrnias I, Thul R, Coombes S, Roderick HL (2011) Atrial cardiomyocyte calcium signalling. Biochim Biophys Acta 1813(5): 922−934 doi: 10.1016/j.bbamcr.2011.01.030
    Bray MA, Geisse NA, Parker KK (2007) Multidimensional detection and analysis of Ca2+ sparks in cardiac myocytes. Biophys J 92(12): 4433−4443 doi: 10.1529/biophysj.106.089359
    Brochet DX, Xie W, Yang D, Cheng H, Lederer WJ (2011) Quarky calcium release in the heart. Circ Res 108(2): 210−218 doi: 10.1161/CIRCRESAHA.110.231258
    Brochet DX, Yang D, Cheng H, Lederer WJ (2012) Elementary calcium release events from the sarcoplasmic reticulum in the heart. Adv Exp Med Biol 740: 499−509
    Cheng H, Lederer WJ (2008) Calcium sparks. Physiol Rev 88(4): 1491−1545 doi: 10.1152/physrev.00030.2007
    Dridi H, Kushnir A, Zalk R, Yuan Q, Melville Z, Marks AR (2020) Intracellular calcium leak in heart failure and atrial fibrillation: a unifying mechanism and therapeutic target. Nat Rev Cardiol 17(11): 732−747 doi: 10.1038/s41569-020-0394-8
    Houser SR (2023) Proarrhythmic remodeling of atrial myocyte Ca2+ handling in atrial fibrillation. JaccBasic Transl Sci 8(1): 16−18
    Huang YF, Lei CX, Xie WJ, Yan L, Wang YR, Yuan S, Wang J, Zhao Y, Wang ZJ, Yang XY, Qin XH, Fang Q, Fang LG, Guo XX (2021) Oxidation of ryanodine receptors promotes Ca2+ leakage and contributes to right ventricular dysfunction in pulmonary hypertension. Hypertension 77(1): 59−71 doi: 10.1161/HYPERTENSIONAHA.120.15561
    Liu T, Xiong F, Qi XY, Xiao JN, Villeneuve L, Abu-Taha I, Dobrev D, Huang CX, Nattel S (2020) Altered calcium handling produces reentry-promoting action potential alternans in atrial fibrillation-remodeled hearts. Jci Insight 5(8): e133754. https://doi.org/10.1172/jci.insight.133754
    Lu FJ, Ma Q, Xie WJ, Liou CL, Zhang DH, Sweat ME, Jardin BD, Naya FJ, Guo YX, Cheng HP, Pu WT (2022) CMYA5 establishes cardiac dyad architecture and positioning. Nat Commun 13(1): 2185. https://doi.org/10.1038/s41467-022-29902-4
    Lu FJ, Zhao Y, Xie WJ, Guo QJ, Wang SQ, Wang XH, Cheng HP (2020) Imaging sarcoplasmic reticulum Ca2+ signaling in intact cardiac myocytes. Circulation 142(15): 1503−1505 doi: 10.1161/CIRCULATIONAHA.120.047784
    Marks AR (2023) Targeting ryanodine receptors to treat human diseases. J Clin Invest 133(2): e162891. https://doi.org/10.1172/JCI162891
    Miotto MC, Weninger G, Dridi H, Yuan Q, Liu Y, Wronska A, Melville Z, Sittenfeld L, Reiken S, Marks AR (2022) Structural analyses of human ryanodine receptor type 2 channels reveal the mechanisms for sudden cardiac death and treatment. Sci Adv 8(29): eabo1272. https://doi.org/10.1126/sciadv.abo1272
    Mohamed BA, Hartmann N, Tirilomis P, Sekeres K, Li W, Neef S, Richter C, Zeisberg EM, Kattner L, Didie M, Guan K, Schmitto JD, Lehnart SE, Luther S, Voigt N, Seidler T, Sossalla S, Hasenfuss G, Toischer K (2018) Sarcoplasmic reticulum calcium leak contributes to arrhythmia but not to heart failure progression. Sci Transl Med 10(458): eaan0724. https://doi.org/10.1126/scitranslmed.aan0724
    Nattel S, Burstein B, Dobrev D (2008) Atrial remodeling and atrial fibrillation mechanisms and implications. Circ Arrhythm Electrophysiol 1(1): 62−73 doi: 10.1161/CIRCEP.107.754564
    Shan J, Xie W, Betzenhauser M, Reiken S, Chen BX, Wronska A, Marks AR (2012) Calcium leak through ryanodine receptors leads to atrial fibrillation in 3 mouse models of catecholaminergic polymorphic ventricular tachycardia. Circ Res 111(6): 708−717 doi: 10.1161/CIRCRESAHA.112.273342
    Shoemaker MB, Yoneda ZT, Crawford DM, Akers WS, Richardson T, Montgomery JA, Phillips S, Shyr Y, Saavedra P, Estrada JC, Kanagasundram A, Shen ST, Michaud GF, Crossley G, Ellis CR, Knollmann BC (2022) A mechanistic clinical trial using (R)- versus (S)-propafenone to test RyR2 (ryanodine receptor) inhibition for the prevention of atrial fibrillation induction. Circ Arrhythm Electrophysiol 15(10): e010713. https://doi.org/10.1161/CIRCEP.121.010713
    Steinberg C, Roston TM, van der Werf C, Sanatani S, Chen SRW, Wilde AAM, Krahn AD (2023) RYR2-ryanodinopathies: from calcium overload to calcium deficiency. Europace 25(6): euad156. https://doi.org/10.1093/europace/euad156
    Szedlak P, Steele DS, Hopkins PM (2023) Cardiac muscle physiology. BJA Educ 23(9): 350−357 doi: 10.1016/j.bjae.2023.05.004
    Tarifa C, Vallmitjana A, Jimenez-Sabado V, Marchena M, Llach A, Herraiz-Martinez A, Godoy-Marin H, Nolla-Colomer C, Ginel A, Vinolas X, Montiel J, Ciruela F, Echebarria B, Benítez R, Cinca J, Hove-Madsen L (2023) Spatial distribution of calcium sparks determines their ability to induce afterdepolarizations in human atrial myocytes. Jacc Basic Transl Sci 8(1): 1−15 doi: 10.1016/j.jacbts.2022.07.013
    Tomek J, Nieves-Cintron M, Navedo MF, Ko CY, Bers DM (2023) SparkMaster 2: a new software for automatic analysis of calcium spark data. Circ Res 133(6): 450−462 doi: 10.1161/CIRCRESAHA.123.322847
    von Wegner F, Both M, Fink RHA (2006) Automated detection of elementary calcium release events using the A Trous wavelet transform. Biophys J 90(6): 2151−2163 doi: 10.1529/biophysj.105.069930
    Walden AP, Dibb KM, Trafford AW (2009) Differences in intracellular calcium homeostasis between atrial and ventricular myocytes. J Mol Cell Cardiol 46(4): 463−473 doi: 10.1016/j.yjmcc.2008.11.003
    Woo SH, Cleemann L, Morad M (2003) Spatiotemporal characteristics of junctional and nonjunctional focal Ca2+ release in rat atrial myocytes. Circ Res 92(1): e1−11 doi: 10.1161/01.RES.0000052925.89426.09
    Xie W, Brochet DX, Wei S, Wang X, Cheng H (2010) Deciphering ryanodine receptor array operation in cardiac myocytes. J Gen Physiol 136(2): 129−133 doi: 10.1085/jgp.201010416
    Xie WJ, Santulli G, Guo XX, Gao M, Chen BX, Marks AR (2013) Imaging atrial arrhythmic intracellular calcium in intact heart. J Mol Cell Cardiol 64: 120−123 doi: 10.1016/j.yjmcc.2013.09.003
    Xie WJ, Santulli G, Reiken SR, Yuan Q, Osborne BW, Chen BX, Marks AR (2015) Mitochondrial oxidative stress promotes atrial fibrillation. Sci Rep 5: 11427. https://doi.org/10.1038/srep11427
    Yang S, Li R, Chen J, Li Z, Huang Z, Xie W (2021) Calcium spark detection and event-based classification of single cardiomyocyte using deep learning. Front Physiol 12: 770051. https://doi.org/10.3389/fphys.2021.770051
    Zhang Y, Qi Y, Li JJ, He WJ, Gao XH, Zhang Y, Sun X, Tong J, Zhang J, Deng XL, Du XJ, Xie W (2021) Stretch-induced sarcoplasmic reticulum calcium leak is causatively associated with atrial fibrillation in pressure-overloaded hearts. Cardiovasc Res 117(4): 1091−1102 doi: 10.1093/cvr/cvaa163
  • 加载中


    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索


    Article Metrics

    Article views (173) PDF downloads(6) Cited by()
    Proportional views


    DownLoad:  Full-Size Img  PowerPoint