培养的HL-1心房肌细胞单层的电压和钙双通道光学映射。
Voltage and calcium dual channel optical mapping of cultured HL-1 atrial myocyte monolayer.
作者信息
Yan Jiajie, Thomson Justin K, Zhao Weiwei, Fast Vladimir G, Ye Tong, Ai Xun
机构信息
Department of Cell and Molecular Physiology, Loyola University Chicago.
Department of Biomedical Engineering, University of Alabama at Birmingham.
出版信息
J Vis Exp. 2015 Mar 23(97):52542. doi: 10.3791/52542.
Abstract
Optical mapping has proven to be a valuable technique to detect cardiac electrical activity on both intact ex vivo hearts and in cultured myocyte monolayers. HL-1 cells have been widely used as a 2-Dimensional cellular model for studying diverse aspects of cardiac physiology. However, it has been a great challenge to optically map calcium (Ca) transients and action potentials simultaneously from the same field of view in a cultured HL-1 atrial cell monolayer. This is because special handling and care is required to prepare healthy cells that can be electrically captured and optically mapped. Therefore, we have developed an optimal working protocol for dual channel optical mapping. In this manuscript, we have described in detail how to perform the dual channel optical mapping experiment. This protocol is a useful tool to enhance the understanding of action potential propagation and Ca kinetics in arrhythmia development.
摘要
光学映射已被证明是一种在完整的离体心脏和培养的心肌细胞单层中检测心脏电活动的有价值技术。HL-1细胞已被广泛用作研究心脏生理学各个方面的二维细胞模型。然而，在培养的HL-1心房细胞单层中，从同一视野同时光学映射钙（Ca）瞬变和动作电位一直是一个巨大的挑战。这是因为需要特殊的处理和护理来制备能够被电捕获和光学映射的健康细胞。因此，我们开发了一种用于双通道光学映射的最佳工作方案。在本手稿中，我们详细描述了如何进行双通道光学映射实验。该方案是增强对心律失常发生过程中动作电位传播和钙动力学理解的有用工具。
相似文献
1
Voltage and calcium dual channel optical mapping of cultured HL-1 atrial myocyte monolayer.培养的HL-1心房肌细胞单层的电压和钙双通道光学映射。
J Vis Exp. 2015 Mar 23(97):52542. doi: 10.3791/52542.
2
Angiotensin II induces complex fractionated electrogram in a cultured atrial myocyte monolayer mediated by calcium and sodium-calcium exchanger.血管紧张素 II 通过钙和钠钙交换体诱导培养的心房心肌细胞单层中的复杂碎裂电图。
Cell Calcium. 2011 Jan;49(1):1-11. doi: 10.1016/j.ceca.2010.10.005. Epub 2010 Dec 17.
3
Characterisation of connexin expression and electrophysiological properties in stable clones of the HL-1 myocyte cell line.HL-1心肌细胞系稳定克隆中连接蛋白表达及电生理特性的表征
PLoS One. 2014 Feb 28;9(2):e90266. doi: 10.1371/journal.pone.0090266. eCollection 2014.
4
A compartmentalized mathematical model of mouse atrial myocytes.一个小鼠心房心肌细胞的房室模型数学模型。
Am J Physiol Heart Circ Physiol. 2020 Mar 1;318(3):H485-H507. doi: 10.1152/ajpheart.00460.2019. Epub 2020 Jan 17.
5
Mechanical stretch of atrial myocyte monolayer decreases sarcoplasmic reticulum calcium adenosine triphosphatase expression and increases susceptibility to repolarization alternans.机械拉伸心房肌细胞单层可降低肌浆网钙三磷酸腺苷酶的表达，并增加复极化交替易感性。
J Am Coll Cardiol. 2011 Nov 8;58(20):2106-15. doi: 10.1016/j.jacc.2011.07.039.
6
Critical roles of a small conductance Ca²⁺-activated K⁺ channel (SK3) in the repolarization process of atrial myocytes.小电导钙激活钾通道（SK3）在心房肌细胞复极化过程中的关键作用。
Cardiovasc Res. 2014 Feb 1;101(2):317-25. doi: 10.1093/cvr/cvt262. Epub 2013 Nov 26.
7
Hypokalemia Promotes Arrhythmia by Distinct Mechanisms in Atrial and Ventricular Myocytes.低钾血症通过不同机制在心房和心室肌细胞中促进心律失常。
Circ Res. 2020 Mar 27;126(7):889-906. doi: 10.1161/CIRCRESAHA.119.315641. Epub 2020 Feb 19.
8
Uncovering the arrhythmogenic potential of TRPM4 activation in atrial-derived HL-1 cells using novel recording and numerical approaches.利用新型记录和数值方法揭示 TRPM4 激活在心房源性 HL-1 细胞中的致心律失常潜能。
Cardiovasc Res. 2017 Aug 1;113(10):1243-1255. doi: 10.1093/cvr/cvx117.
9
Expression changes of ionic channels in early phase of cultured rat atrial myocytes induced by rapid pacing.快速起搏诱导的培养大鼠心房肌细胞早期离子通道的表达变化
J Cardiothorac Surg. 2013 Sep 29;8:194. doi: 10.1186/1749-8090-8-194.
10
Age-related changes in cardiac electrophysiology and calcium handling in response to sympathetic nerve stimulation.与年龄相关的心脏电生理学和钙处理变化对交感神经刺激的反应。
J Physiol. 2018 Sep;596(17):3977-3991. doi: 10.1113/JP276396. Epub 2018 Aug 3.
引用本文的文献
1
Long-Term Alcohol-Activated c-Jun N-terminal Kinase Isoform 2 Preserves Cardiac Function but Drives Ca-Triggered Arrhythmias.长期激活的 c-Jun N 末端激酶同工型 2 保护心脏功能，但引发钙触发的心律失常。
Cells. 2023 Sep 8;12(18):2233. doi: 10.3390/cells12182233.
2
Molecular remodeling of Cx43, but not structural remodeling, promotes arrhythmias in an arrhythmogenic canine model of nonischemic heart failure.缝隙连接蛋白 43 的分子重构而非结构重构促进非缺血性心力衰竭致心律失常犬模型的心律失常。
J Mol Cell Cardiol. 2021 Sep;158:72-81. doi: 10.1016/j.yjmcc.2021.05.012. Epub 2021 May 31.
3
Characterisation of re-entrant circuit (or rotational activity) in vitro using the HL1-6 myocyte cell line.使用 HL1-6 心肌细胞系在体表征折返环（或旋转活动）。
J Mol Cell Cardiol. 2018 Jun;119:155-164. doi: 10.1016/j.yjmcc.2018.05.002. Epub 2018 May 7.
4
Transcriptional regulation of stress kinase JNK2 in pro-arrhythmic CaMKIIδ expression in the aged atrium.转录调控应激激酶 JNK2 在老年心房促心律失常性 CaMKIIδ 表达中的作用。
Cardiovasc Res. 2018 Apr 1;114(5):737-746. doi: 10.1093/cvr/cvy011.
5
Stress Signaling JNK2 Crosstalk With CaMKII Underlies Enhanced Atrial Arrhythmogenesis.应激信号转导 JNK2 与 CaMKII 的串扰是增强心房心律失常发生的基础。
Circ Res. 2018 Mar 16;122(6):821-835. doi: 10.1161/CIRCRESAHA.117.312536. Epub 2018 Jan 19.
6
The stress kinase JNK regulates gap junction Cx43 gene expression and promotes atrial fibrillation in the aged heart.应激激酶 JNK 调节缝隙连接蛋白 Cx43 基因表达并促进老年心脏中的心房颤动。
J Mol Cell Cardiol. 2018 Jan;114:105-115. doi: 10.1016/j.yjmcc.2017.11.006. Epub 2017 Nov 13.
本文引用的文献
1
c-Jun N-terminal kinase activation contributes to reduced connexin43 and development of atrial arrhythmias.c-Jun N-末端激酶的激活导致连接蛋白 43 减少和房性心律失常的发生。
Cardiovasc Res. 2013 Mar 1;97(3):589-97. doi: 10.1093/cvr/cvs366. Epub 2012 Dec 14.
2
Ionic mechanism of shock-induced arrhythmias: role of intracellular calcium.休克诱导心律失常的离子机制：细胞内钙的作用。
Heart Rhythm. 2012 Jan;9(1):96-104. doi: 10.1016/j.hrthm.2011.08.024. Epub 2011 Aug 28.
3
A major role for HERG in determining frequency of reentry in neonatal rat ventricular myocyte monolayer.HERG 在新生大鼠心室肌细胞单层中决定折返频率的主要作用。
Circ Res. 2010 Dec 10;107(12):1503-11. doi: 10.1161/CIRCRESAHA.110.232470. Epub 2010 Oct 14.
4
Simultaneous optical mapping of intracellular free calcium and action potentials from Langendorff perfused hearts.对Langendorff灌注心脏的细胞内游离钙和动作电位进行同步光学映射。
Curr Protoc Cytom. 2009 Jul;Chapter 12:Unit 12.17. doi: 10.1002/0471142956.cy1217s49.
5
Decreased intercellular coupling improves the function of cardiac pacemakers derived from mouse embryonic stem cells.细胞间偶联的减少改善了源自小鼠胚胎干细胞的心脏起搏器的功能。
J Mol Cell Cardiol. 2008 Nov;45(5):642-9. doi: 10.1016/j.yjmcc.2008.08.013. Epub 2008 Sep 11.
6
Electrogram fractionation in murine HL-1 atrial monolayer model.小鼠HL-1心房单层模型中的电信号碎裂
Heart Rhythm. 2008 Jul;5(7):1029-35. doi: 10.1016/j.hrthm.2008.03.022. Epub 2008 Mar 21.
7
Relative contributions of connexins 40 and 43 to atrial impulse propagation in synthetic strands of neonatal and fetal murine cardiomyocytes.连接蛋白40和43对新生和胎儿小鼠心肌细胞合成束中房性冲动传导的相对贡献。
Circ Res. 2006 Nov 24;99(11):1216-24. doi: 10.1161/01.RES.0000250607.34498.b4. Epub 2006 Oct 19.
8
Simultaneous optical imaging of membrane potential and intracellular calcium.膜电位与细胞内钙的同步光学成像。
J Electrocardiol. 2005 Oct;38(4 Suppl):107-12. doi: 10.1016/j.jelectrocard.2005.06.023.
9
Rapid stimulation causes electrical remodeling in cultured atrial myocytes.快速刺激可导致培养的心房肌细胞发生电重构。
J Mol Cell Cardiol. 2005 Feb;38(2):299-308. doi: 10.1016/j.yjmcc.2004.11.015. Epub 2005 Jan 20.
10
Optical imaging of the heart.心脏的光学成像。
Circ Res. 2004 Jul 9;95(1):21-33. doi: 10.1161/01.RES.0000130529.18016.35.
Zotero 插件