Department of Physics and Computer Science, Dayalbagh Educational Institute, Agra, 282005, India.
Pflugers Arch. 2023 Dec;475(12):1479-1503. doi: 10.1007/s00424-023-02831-x. Epub 2023 Jul 7.
A major challenge in cardiac optogenetics is to have minimally invasive large volume excitation and suppression for effective cardioversion and treatment of tachycardia. It is important to study the effect of light attenuation on the electrical activity of cells in in vivo cardiac optogenetic experiments. In this computational study, we present a detailed analysis of the effect of light attenuation in different channelrhodopsins (ChRs)-expressing human ventricular cardiomyocytes. The study shows that sustained illumination from the myocardium surface used for suppression, simultaneously results in spurious excitation in deeper tissue regions. Tissue depths of suppressed and excited regions have been determined for different opsin expression levels. It is shown that increasing the expression level by 5-fold enhances the depth of suppressed tissue from 2.24 to 3.73 mm with ChR2(H134R) (ChR2 with a single point mutation at position H134), 3.78 to 5.12 mm with GtACR1 (anion-conducting ChR from cryptophyte algae Guillardia theta) and 6.63 to 9.31 mm with ChRmine (a marine opsin gene from Tiarina fusus). Light attenuation also results in desynchrony in action potentials in different tissue regions under pulsed illumination. It is further shown that gradient-opsin expression not only enables suppression up to the same level of tissue depth but also enables synchronized excitation under pulsed illumination. The study is important for the effective treatment of tachycardia and cardiac pacing and for extending the scale of cardiac optogenetics.
心脏光遗传学的一个主要挑战是实现微创的大容量激发和抑制,以有效进行心脏复律和治疗心动过速。在体内心脏光遗传学实验中,研究光衰减对细胞电活动的影响非常重要。在这项计算研究中,我们对不同通道蛋白(ChR)表达的人心室肌细胞中光衰减的影响进行了详细分析。研究表明,用于抑制的心肌表面持续照射,同时会导致深层组织区域出现虚假兴奋。不同光感受器表达水平下,抑制和兴奋区域的组织深度已经确定。结果表明,ChR2(H134R)(在位置 H134 处具有单点突变的 ChR2)的表达水平提高 5 倍,可将抑制组织的深度从 2.24 增加到 3.73mm,GtACR1(隐藻的阴离子传导 ChR)可从 3.78 增加到 5.12mm,ChRmine(来自 Tiarina fusus 的海洋光感受器基因)可从 6.63 增加到 9.31mm;用 ChR2(H134R)表达水平提高 5 倍,可将抑制组织的深度从 2.24 增加到 3.73mm,GtACR1(隐藻的阴离子传导 ChR)可从 3.78 增加到 5.12mm,ChRmine(来自 Tiarina fusus 的海洋光感受器基因)可从 6.63 增加到 9.31mm;用 ChR2(H134R)表达水平提高 5 倍,可将抑制组织的深度从 2.24 增加到 3.73mm,GtACR1(隐藻的阴离子传导 ChR)可从 3.78 增加到 5.12mm,ChRmine(来自 Tiarina fusus 的海洋光感受器基因)可从 6.63 增加到 9.31mm;用 ChRmine 表达水平提高 5 倍,可将抑制组织的深度从 6.63 增加到 9.31mm。光衰减还会导致在脉冲照射下不同组织区域的动作电位失步。进一步表明,梯度光感受器表达不仅能使抑制达到相同的组织深度,还能使脉冲照射下的兴奋同步。该研究对心动过速和心脏起搏的有效治疗以及心脏光遗传学的扩展具有重要意义。