Laboratory of Developmental Biology, Department of Biology and Biotechnology "Lazzaro Spallanzani", University of Pavia, Via Ferrata 9, 27100, Pavia, Italy.
Department of Electrical, Computer and Biomedical Engineering (DIII), University of Pavia, Via Ferrata 5, Pavia, Italy.
Sci Rep. 2021 Nov 30;11(1):23116. doi: 10.1038/s41598-021-02590-8.
Chronic exposure to environmental pollutants threatens human health. Arsenic, a world-wide diffused toxicant, is associated to cardiac pathology in the adult and to congenital heart defects in the foetus. Poorly known are its effects on perinatal cardiomyocytes. Here, bioinformatic image-analysis tools were coupled with cellular and molecular analyses to obtain functional and structural quantitative metrics of the impairment induced by 0.1, 0.5 or 1.0 µM arsenic trioxide exposure on the perinatal-like cardiomyocyte component of mouse embryoid bodies, within their 3D complex cell organization. With this approach, we quantified alterations to the (a) beating activity; (b) sarcomere organization (texture, edge, repetitiveness, height and width of the Z bands); (c) cardiomyocyte size and shape; (d) volume occupied by cardiomyocytes within the EBs. Sarcomere organization and cell morphology impairment are paralleled by differential expression of sarcomeric α-actin and Tropomyosin proteins and of acta2, myh6 and myh7 genes. Also, significant increase of Cx40, Cx43 and Cx45 connexin genes and of Cx43 protein expression profiles is paralleled by large Cx43 immunofluorescence signals. These results provide new insights into the role of arsenic in impairing cytoskeletal components of perinatal-like cardiomyocytes which, in turn, affect cell size, shape and beating capacity.
慢性暴露于环境污染物会威胁人类健康。砷是一种在全球范围内广泛存在的有毒物质，它与成年人的心脏病理学以及胎儿的先天性心脏缺陷有关。其对围产期心肌细胞的影响知之甚少。在这里，生物信息学图像分析工具与细胞和分子分析相结合，获得了 0.1、0.5 或 1.0µM 三氧化二砷暴露对其 3D 复杂细胞组织中的小鼠胚体类心肌细胞成分的功能和结构定量指标的损伤。通过这种方法，我们定量了(a)搏动活性；(b)肌节组织（纹理、边缘、重复性、Z 带的高度和宽度）；(c)心肌细胞大小和形状；(d)心肌细胞在 EB 中所占的体积。肌节组织和细胞形态损伤与肌节α-肌动蛋白和原肌球蛋白蛋白以及 acta2、myh6 和 myh7 基因的差异表达相平行。此外，Cx40、Cx43 和 Cx45 连接蛋白基因和 Cx43 蛋白表达谱的显著增加与 Cx43 免疫荧光信号的增大相平行。这些结果为砷在损伤围产期样心肌细胞的细胞骨架成分方面提供了新的见解，进而影响细胞大小、形状和搏动能力。
