title: Atomic force microscopy and high-content analysis: two innovative technologies for dissecting the relationship between epithelial-mesenchymal transition-related morphological and structural alterations and cell mechanical properties.
abstract: Epithelial-mesenchymal transition (EMT) is a complex series of cellular reprogramming events culminating in striking alterations in morphology towards an invasive mesenchymal phenotype. Increasingly, evidence suggests that EMT exerts a pivotal role in pathophysiological situations including fibrosis and cancer. Core to these dynamical changes in cellular polarity and plasticity is discrete modifications in cytoskeletal structure. In particular, newly established actin-stress fibres supplant a preceding system of highly organised cortical actin. Although cumulative studies have contributed to elucidation of the detailed signalling pathways that underpin this elaborate molecular process, there remains a deficiency regarding its precise contribution to cellular biomechanics. The advent of atomic force microscopy (AFM) and high-content analysis (HCA) provides two innovative technologies for dissecting the relationship between EMT-related morphological and structural alterations and cell mechanical properties. AFM permits acquisition of high resolution topographical images and detailed analysis of cellular viscoelasticity while HCA facilitates a comprehensive and perspicacious assessment of morphological changes. In combination, they offer the possibility of novel insights into the dynamic traits of transitioning cells. Herein, a detailed protocol describing AFM and HCA techniques for evaluation of transforming growth factor-β1-induced EMT of alveolar epithelial cells is provided.
authors: School of Pharmacy & Pharmaceutical Sciences, Trinity College Dublin, Ireland. bucklest@tcd.ie
authors: Stephen T
authors: ST
authors: Buckley
authors: Anthony M
authors: AM
authors: Davies
authors: Carsten
authors: C
authors: Ehrhardt
mesh_terms: D054337:Cell Dedifferentiation / Q000235:genetics; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000648:ultrastructure; D004847:Epithelial Cells / Q000648:ultrastructure; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005355:Fibrosis / Q000235:genetics; D006801:Humans; D008648:Mesoderm / Q000648:ultrastructure*; D018625:Microscopy, Atomic Force / Q000379:methods*; D009369:Neoplasms / Q000235:genetics; D011650:Pulmonary Alveoli / Q000648:ultrastructure; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D016212:Transforming Growth Factor beta