OptoGap is an optogenetics-enabled assay for quantification of cell-cell coupling in multicellular cardiac tissue.
OptoGap is an optogenetics-enabled assay for quantification of cell-cell coupling in multicellular cardiac tissue.
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DOI：
10.1038/s41598-021-88573-1
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发表时间：
2021-04-29
影响因子：
4.6
通讯作者：
Entcheva E
中科院分区：
文献类型：
--
作者：
Boyle PM;Yu J;Klimas A;Williams JC;Trayanova NA;Entcheva E
细胞间的电耦合是细胞间通讯的重要手段。例如，当心肌成纤维细胞和心肌细胞之间的病理性电耦合导致心律失常风险增加时，或者在细胞治疗方法中供体(例如，心脏祖细胞)细胞与天然心肌细胞整合时，获得心肌细胞和不可兴奋细胞之间的这种偶联的定量知识是很重要的。目前，还没有直接的方法来评估多细胞组织内的异细胞偶联。在这里，我们通过实验和计算演示了一种新的非接触式电耦合测试，OptoGap，基于对表达光遗传致动器的不可兴奋细胞的选择性照明，以及对耦合的可兴奋细胞(例如，心肌细胞)的光不敏感响应的光学传感。细胞间的耦合是通过光刺激细胞的连接电流产生动作电位所需的能量来量化的(S)。所提出的技术与标准的间接方法GapFRAP进行了实验验证，该方法使用二维环境中的光敏心脏成纤维细胞和未转化的心肌细胞。计算模型和适当的校准证实了它对复杂的本征心脏三维环境的潜在适用性。最后，通过计算分析了OptoGap对固有细胞尺度兴奋性的敏感性。
Intercellular electrical coupling is an essential means of communication between cells. It is important to obtain quantitative knowledge of such coupling between cardiomyocytes and non-excitable cells when, for example, pathological electrical coupling between myofibroblasts and cardiomyocytes yields increased arrhythmia risk or during the integration of donor (e.g., cardiac progenitor) cells with native cardiomyocytes in cell-therapy approaches. Currently, there is no direct method for assessing heterocellular coupling within multicellular tissue. Here we demonstrate experimentally and computationally a new contactless assay for electrical coupling, OptoGap, based on selective illumination of inexcitable cells that express optogenetic actuators and optical sensing of the response of coupled excitable cells (e.g., cardiomyocytes) that are light-insensitive. Cell–cell coupling is quantified by the energy required to elicit an action potential via junctional current from the light-stimulated cell(s). The proposed technique is experimentally validated against the standard indirect approach, GapFRAP, using light-sensitive cardiac fibroblasts and non-transformed cardiomyocytes in a two-dimensional setting. Its potential applicability to the complex three-dimensional setting of the native heart is corroborated by computational modelling and proper calibration. Lastly, the sensitivity of OptoGap to intrinsic cell-scale excitability is robustly characterized via computational analysis.
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影响因子：
4
作者：
Boyle PM;Hakim JB;Zahid S;Franceschi WH;Murphy MJ;Vigmond EJ;Dubois R;Haïssaguerre M;Hocini M;Jaïs P;Trayanova NA;Cochet H
通讯作者：
Cochet H
DOI：
10.1016/j.pbiomolbio.2014.07.001
发表时间：
2014-08
影响因子：
3.8
作者：
Ambrosi CM;Klimas A;Yu J;Entcheva E
通讯作者：
Entcheva E
影响因子：
20.1
作者：
Ashihara T;Haraguchi R;Nakazawa K;Namba T;Ikeda T;Nakazawa Y;Ozawa T;Ito M;Horie M;Trayanova NA
通讯作者：
Trayanova NA
影响因子：
4
作者：
Boyle PM;Hakim JB;Zahid S;Franceschi WH;Murphy MJ;Prakosa A;Aronis KN;Zghaib T;Balouch M;Ipek EG;Chrispin J;Berger RD;Ashikaga H;Marine JE;Calkins H;Nazarian S;Spragg DD;Trayanova NA
通讯作者：
Trayanova NA
DOI：
10.1161/circep.111.964247
发表时间：
2011-10
期刊：
Circulation. Arrhythmia and electrophysiology
影响因子：
--
作者：
Jia Z;Valiunas V;Lu Z;Bien H;Liu H;Wang HZ;Rosati B;Brink PR;Cohen IS;Entcheva E
通讯作者：
Entcheva E