Properties of blebbistatin for cardiac optical mapping and other imaging applications.
Properties of blebbistatin for cardiac optical mapping and other imaging applications.
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肌球蛋白在心脏光学测绘和其他成像应用中的特性。
DOI：
10.1007/s00424-012-1147-2
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Sarvazyan,Narine
中科院分区：
文献类型：
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作者：
Swift,LutherM;Asfour,Huda;Posnack,NikkiG;Arutunyan,Ara;Kay,MatthewW;Sarvazyan,Narine
Blebbistatin 是最近发现的一种肌球蛋白 II 抑制剂。它正迅速成为减少心脏光学测绘过程中运动伪影以及研究细胞运动和细胞侵袭的首选化合物。尽管肌球蛋白与其他机电解偶联剂相比具有许多优点，但其许多特性仍有待解决。在这里，我们描述了与肌球蛋白的使用相关的几个方法学问题，包括其光谱特性、可逆性及其对组织代谢状态的影响。我们发现，如果不采取预防措施，用肌球蛋白灌注可能会导致肌球蛋白沉淀在脉管系统中积聚。尽管此类沉淀物具有荧光性，但在通常用于跨膜电压荧光成像的波段（即发射波长 >600 nm）内无法检测到它。因此，在使用电压敏感染料的研究中，布雷他汀阻断微循环可能会导致数据误解。布雷他汀还可能影响绿色荧光团的成像，因为它会引起内源性组织荧光的光谱偏移。沉淀物形式和各种溶液（DMSO、水和 1% 水性白蛋白）中的肌球蛋白 3D 激发-发射矩阵揭示了该分子在不同环境中荧光的显着变化。最后，我们检查了blebbistatin对心脏收缩的解偶联作用的可逆性。我们的研究结果提供了有关这种肌球蛋白 II 抑制剂特性的重要新信息，这将有助于正确设计和解释使用该化合物的研究。
Blebbistatin is a recently discovered myosin II inhibitor. It is rapidly becoming a compound of choice to reduce motion artifacts during cardiac optical mapping, as well as to study cell motility and cell invasion. Although blebbistatin has a number of advantages over other electromechanical uncouplers, many of its properties have yet to be addressed. Here we describe several methodological issues associated with the use of blebbistatin, including its spectral properties, reversibility, and its effect on tissue metabolic state. We show that if precautions are not taken, perfusion with blebbistatin may result in blebbistatin precipitate that accumulates in the vasculature. Although such precipitate is fluorescent, it is not detectable within wavelength bands that are typically used for transmembrane voltage fluorescence imaging (i.e., emission wavelengths >600 nm). Therefore, blockage of the microcirculation by blebbistatin may cause data misinterpretation in studies that use voltage-sensitive dyes. Blebbistatin may also impact imaging of green fluorophores due to the spectral shift it causes in endogenous tissue fluorescence. 3D excitation–emission matrices of blebbistatin in precipitate form and in various solutions (DMSO, water, and 1 % aqueous albumin) revealed significant changes in the fluorescence of this molecule in different environments. Finally, we examined the reversibility of blebbistatin's uncoupling effect on cardiac contraction. Our findings provide important new information about the properties of this myosin II inhibitor, which will aid in the proper design and interpretation of studies that use this compound.