A Protocol for Transverse Cardiac Slicing and Optical Mapping in Murine Heart
A Protocol for Transverse Cardiac Slicing and Optical Mapping in Murine Heart
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小鼠心脏横向心脏切片和光学映射的协议
DOI：
10.3389/fphys.2019.00755
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发表时间：
2019-06-25
影响因子：
4
通讯作者：
Lei, M.
中科院分区：
文献类型：
--
作者：
He, S.;Wen, Q.;Lei, M.
薄的活体组织切片是近年来出现的一种用于心脏电生理研究的新的组织模型。除了小型和大型哺乳动物心脏外，切片还可以从人类心脏组织中产生，代表了用于临床前和转化心脏研究的强大体外模型系统。在本协议中，我们描述了一个详细的小鼠心脏横向切片和光学成像方法。使用这种技术的高通量光学成像允许在器官型伪二维模型中研究小鼠心脏的电生理学。切片以与心脏长轴成直角的方式切割，允许以出色的区域精度对整个心脏的跨膜电位（Vm）和钙瞬变（CaT）进行稳健的询问。该方法使得能够使用从心室制备的一系列切片来以高时间和空间分辨率测量Vm和CaT，从而允许（i）比较形成代表心脏的原始几何形状的堆叠的连续切片;（ii）描绘心室中的Vm和CaT的跨壁和区域梯度;（iii）在应激下动作电位和CaT交替的跨壁和区域概况的表征（例如，高频起搏或β-肾上腺素能刺激）或病理状况（例如，肥大）。因此，这里描述的协议提供了一个强大的平台，心脏内的电和钙处理异质性的创新研究。它还可以与光遗传学技术相结合，以进行光刺激;以细胞类型特异性的方式辅助细胞Vm和CaT的研究。
Thin living tissue slices have recently emerged as a new tissue model for cardiac electrophysiological research. Slices can be produced from human cardiac tissue, in addition to small and large mammalian hearts, representing a powerful in vitro model system for preclinical and translational heart research. In the present protocol, we describe a detailed mouse heart transverse slicing and optical imaging methodology. The use of this technology for high-throughput optical imaging allows study of electrophysiology of murine hearts in an organotypic pseudo two-dimensional model. The slices are cut at right angles to the long axis of the heart, permitting robust interrogation of transmembrane potential (Vm) and calcium transients (CaT) throughout the entire heart with exceptional regional precision. This approach enables the use of a series of slices prepared from the ventricles to measure Vm and CaT with high temporal and spatial resolution, allowing (i) comparison of successive slices which form a stack representing the original geometry of the heart; (ii) profiling of transmural and regional gradients in Vm and CaT in the ventricle; (iii) characterization of transmural and regional profiles of action potential and CaT alternans under stress (e.g., high frequency pacing or β-adrenergic stimulation) or pathological conditions (e.g., hypertrophy). Thus, the protocol described here provides a powerful platform for innovative research on electrical and calcium handling heterogeneity within the heart. It can be also combined with optogenetic technology to carry out optical stimulation; aiding studies of cellular Vm and CaT in a cell type specific manner.