Borin Daniele, Peña Brisa, Chen Suet Nee, Long Carlin S, Taylor Matthew R G, Mestroni Luisa, Sbaizero Orfeo
Department of Engineering and Architecture, University of Trieste, Trieste, Italy.
Cardiovascular Institute, University of Colorado Anschutz Medical Campus - Aurora, Co., USA.
Heliyon. 2020 Jan 23;6(1):e03175. doi: 10.1016/j.heliyon.2020.e03175. eCollection 2020 Jan.
Given the clinical effect of laminopathies, understanding lamin mechanical properties will benefit the treatment of heart failure. Here we report a mechano-dynamic study of mutations in neonatal rat ventricular myocytes (NRVM) using single cell spectroscopy with Atomic Force Microscopy (AFM) and measured changes in beating force, frequency and contractile amplitude of selected mutant-expressing cells within cell clusters. Furthermore, since beat-to-beat variations can provide clues on the origin of arrhythmias, we analyzed the beating rate variability using a time-domain method which provides a Poincaré plot. Data were further correlated to cell phenotypes. Immunofluorescence and calcium imaging analysis showed that mutant lamin changed NRVMs beating force and frequency. Additionally, we noted an altered microtubule network organization with shorter filament length, and defective hemichannel membrane localization (Connexin 43). These data highlight the interconnection between nucleoskeleton, cytoskeleton and sarcolemmal structures, and the transcellular consequences of mutant lamin protein in the pathogenesis of the cardiac .
鉴于核纤层蛋白病的临床效应，了解核纤层蛋白的力学特性将有助于心力衰竭的治疗。在此，我们报告了一项对新生大鼠心室肌细胞（NRVM）突变体的机械动力学研究，该研究使用原子力显微镜（AFM）进行单细胞光谱分析，并测量了细胞簇中选定的表达突变体的细胞的搏动力、频率和收缩幅度的变化。此外，由于逐搏变化可为心律失常的起源提供线索，我们使用提供庞加莱图的时域方法分析了搏动率变异性。数据进一步与细胞表型相关。免疫荧光和钙成像分析表明，突变型核纤层蛋白改变了NRVM的搏动力和频率。此外，我们注意到微管网络组织发生改变，细丝长度变短，半通道膜定位（连接蛋白43）存在缺陷。这些数据突出了核骨架、细胞骨架和肌膜结构之间的相互联系，以及突变型核纤层蛋白在心脏发病机制中的跨细胞影响。
