Imperial College London, Du Cane road, W12 0NN, London, UK.
Department of Biomedical Engineering, George Washington University, Washington, DC, USA.
Commun Biol. 2023 Nov 8;6(1):1131. doi: 10.1038/s42003-023-05509-3.
We present a novel framework, Opto-SICM, for studies of cellular interactions in live cells with high spatiotemporal resolution. The approach combines scanning ion conductance microscopy, SICM, and cell-type-specific optogenetic interrogation. Light-excitable cardiac fibroblasts (FB) and myofibroblasts (myoFB) were plated together with non-modified cardiomyocytes (CM) and then paced with periodic illumination. Opto-SICM reveals the extent of FB/myoFB-CM cell-cell contacts and the dynamic changes over time not visible by optical microscopy. FB-CM pairs have lower gap junctional expression of connexin-43 and higher contact dynamism compared to myoFB-CM pairs. The responsiveness of CM to pacing via FB/myoFB depends on the dynamics of the contact but not on the area. The non-responding pairs have higher net cell-cell movement at the contact. These findings are relevant to cardiac disease states, where adverse remodeling leads to abnormal electrical excitation of CM. The Opto-SICM framework can be deployed to offer new insights on cellular and subcellular interactions in various cell types, in real-time.
我们提出了一种新的框架，即光刺激扫描离子电导显微镜（Opto-SICM），用于以高时空分辨率研究活细胞中的细胞相互作用。该方法结合了扫描离子电导显微镜（SICM）和细胞类型特异性光遗传学询问。光可兴奋的心脏成纤维细胞（FB）和肌成纤维细胞（myoFB）与未修饰的心肌细胞（CM）一起铺板，然后用周期性光照进行起搏。Opto-SICM 揭示了 FB/myoFB-CM 细胞-细胞接触的程度以及通过光学显微镜无法观察到的随时间的动态变化。与 myoFB-CM 对相比，FB-CM 对具有更低的缝隙连接蛋白 43 的表达和更高的接触动力学。CM 通过 FB/myoFB 起搏的反应性取决于接触的动力学而不是面积。非反应性对在接触处具有更高的净细胞间运动。这些发现与心脏疾病状态有关，其中不良重塑导致 CM 的异常电兴奋。Opto-SICM 框架可用于实时提供关于各种细胞类型中细胞和亚细胞相互作用的新见解。
