Children's National Heart Institute, Children's National Hospital, Washington, District of Columbia.
Sheikh Zayed Institute for Pediatric Surgical Innovation, Children's National Hospital, Washington, District of Columbia.
Am J Physiol Heart Circ Physiol. 2021 Dec 1;321(6):H1005-H1013. doi: 10.1152/ajpheart.00477.2021. Epub 2021 Oct 8.
Optical mapping is an imaging technique that is extensively used in cardiovascular research, wherein parameter-sensitive fluorescent indicators are used to study the electrophysiology and excitation-contraction coupling of cardiac tissues. Despite many benefits of optical mapping, eliminating motion artifacts within the optical signals is a major challenge, as myocardial contraction interferes with the faithful acquisition of action potentials and intracellular calcium transients. As such, excitation-contraction uncoupling agents are frequently used to reduce signal distortion by suppressing contraction. When compared with other uncoupling agents, blebbistatin is the most frequently used, as it offers increased potency with minimal direct effects on cardiac electrophysiology. Nevertheless, blebbistatin may exert secondary effects on electrical activity, metabolism, and coronary flow, and the incorrect administration of blebbistatin to cardiac tissue can prove detrimental, resulting in erroneous interpretation of optical mapping results. In this "Getting It Right" perspective, we briefly review the literature regarding the use of blebbistatin in cardiac optical mapping experiments, highlight potential secondary effects of blebbistatin on cardiac electrical activity and metabolic demand, and conclude with the consensus of the authors on best practices for effectively using blebbistatin in optical mapping studies of cardiac tissue.
光学绘图是一种广泛应用于心血管研究的成像技术，其中使用参数敏感的荧光指示剂来研究心脏组织的电生理和兴奋-收缩偶联。尽管光学绘图有许多优点，但消除光学信号中的运动伪影是一个主要挑战，因为心肌收缩会干扰动作电位和细胞内钙瞬变的准确采集。因此，经常使用兴奋-收缩偶联解偶联剂来减少收缩引起的信号失真。与其他解偶联剂相比，blebbistatin 是最常用的，因为它在对心脏电生理的影响最小的情况下提供了更高的效力。然而，blebbistatin 可能对电活动、代谢和冠状动脉血流产生继发性影响，并且不正确地将 blebbistatin 施用于心脏组织可能会产生有害影响，导致对光学绘图结果的错误解释。在这篇“正确应用”的观点中，我们简要回顾了关于在心脏光学绘图实验中使用 blebbistatin 的文献，强调了 blebbistatin 对心脏电活动和代谢需求的潜在继发性影响，并总结了作者对有效使用 blebbistatin 进行心脏组织光学绘图研究的最佳实践的共识。
