SFAlab: image-based quantification of mechano-active ventral actin stress fibers in adherent cells

Frontiers in Cell and Developmental Biology

Cell and Developmental Biology

Mark C. van Turnhout¹  Bart G. W. Groenen¹  Dylan Mostert²  Nicholas A. Kurniawan²  Vito Conte²  Carlijn V. C. Bouten²  Janine Grolleman²  Cecilia M. Sahlgren³ 

[1] Department of Biomedical Engineering, Laboratory for Cell and Tissue Engineering, Eindhoven University of Technology, Eindhoven, Netherlands;Department of Biomedical Engineering, Laboratory for Cell and Tissue Engineering, Eindhoven University of Technology, Eindhoven, Netherlands;Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven, Netherlands;Department of Biomedical Engineering, Laboratory for Cell and Tissue Engineering, Eindhoven University of Technology, Eindhoven, Netherlands;Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven, Netherlands;Faculty of Science and Engineering, Åbo Akademi University, Turku, Finland;

关键词: ventral stress fibers;    mechanobiology;    microscopy;    quantitative image analysis;    adherent cells;   

DOI  :  10.3389/fcell.2023.1267822

 received in 2023-07-27, accepted in 2023-09-06,  发布年份 2023

来源: Frontiers

【 摘 要 】

Ventral actin stress fibers (SFs) are a subset of actin SFs that begin and terminate at focal adhesion (FA) complexes. Ventral SFs can transmit forces from and to the extracellular matrix and serve as a prominent mechanosensing and mechanotransduction machinery for cells. Therefore, quantitative analysis of ventral SFs can lead to deeper understanding of the dynamic mechanical interplay between cells and their extracellular matrix (mechanoreciprocity). However, the dynamic nature and organization of ventral SFs challenge their quantification, and current quantification tools mainly focus on all SFs present in cells and cannot discriminate between subsets. Here we present an image analysis-based computational toolbox, called SFAlab, to quantify the number of ventral SFs and the number of ventral SFs per FA, and provide spatial information about the locations of the identified ventral SFs. SFAlab is built as an all-in-one toolbox that besides analyzing ventral SFs also enables the identification and quantification of (the shape descriptors of) nuclei, cells, and FAs. We validated SFAlab for the quantification of ventral SFs in human fetal cardiac fibroblasts and demonstrated that SFAlab analysis i) yields accurate ventral SF detection in the presence of image imperfections often found in typical fluorescence microscopy images, and ii) is robust against user subjectivity and potential experimental artifacts. To demonstrate the usefulness of SFAlab in mechanobiology research, we modulated actin polymerization and showed that inhibition of Rho kinase led to a significant decrease in ventral SF formation and the number of ventral SFs per FA, shedding light on the importance of the RhoA pathway specifically in ventral SF formation. We present SFAlab as a powerful open source, easy to use image-based analytical tool to increase our understanding of mechanoreciprocity in adherent cells.

【 授权许可】

Unknown   
Copyright © 2023 Mostert, Grolleman, van Turnhout, Groenen, Conte, Sahlgren, Kurniawan and Bouten.