Acute effects of cell stretch on keratin filaments in A549 lung cells

Lutz, Anngrit; Jung, Dominik; Diem, Kathrin; Fauler, Michael; Port, Fabian; Gottschalk, Kay E.; Felder, Edward

doi:10.1096/fj.201903160rr

Cited by 5 publications

(4 citation statements)

References 57 publications

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“…2 B ). Consistent with previous studies ( Felder et al, 2008 ; Lutz et al, 2020 ), our quantitative analysis revealed that monolayer stretching led to a significant increase in the cross-sectional DSM width in both WT and KO monolayers ( Fig. 2 E ).…”

Section: Resultssupporting

confidence: 92%

Plectin-mediated cytoskeletal crosstalk controls cell tension and cohesion in epithelial sheets

Přechová

Adamova

Schweizer

et al. 2022

Journal of Cell Biology

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The coordinated interplay of cytoskeletal networks critically determines tissue biomechanics and structural integrity. Here, we show that plectin, a major intermediate filament-based cytolinker protein, orchestrates cortical cytoskeletal networks in epithelial sheets to support intercellular junctions. By combining CRISPR/Cas9-based gene editing and pharmacological inhibition, we demonstrate that in an F-actin–dependent context, plectin is essential for the formation of the circumferential keratin rim, organization of radial keratin spokes, and desmosomal patterning. In the absence of plectin-mediated cytoskeletal cross-linking, the aberrant keratin–desmosome (DSM)–network feeds back to the actin cytoskeleton, which results in elevated actomyosin contractility. Also, by complementing a predictive mechanical model with Förster resonance energy transfer–based tension sensors, we provide evidence that in the absence of cytoskeletal cross-linking, major intercellular junctions (adherens junctions and DSMs) are under intrinsically generated tensile stress. Defective cytoarchitecture and tensional disequilibrium result in reduced intercellular cohesion, associated with general destabilization of plectin-deficient sheets upon mechanical stress.

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Section: Resultssupporting

confidence: 92%

Plectin-mediated cytoskeletal crosstalk controls cell tension and cohesion in epithelial sheets

Přechová

Adamova

Schweizer

et al. 2022

Journal of Cell Biology

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“…Using Equation (1), those curves can be fitted to calculate the Young's modulus at each position. AFM has been used to study the mechanics of different cell types [114][115][116] and has helped in understanding the underlying processes in various diseases, including cancer [117,118].…”

Section: Active Cellular Force Probingmentioning

confidence: 99%

“…AFM has been used to study the mechanics of different cell types [ 114 , 115 , 116 ] and has helped in understanding the underlying processes in various diseases, including cancer [ 117 , 118 ].…”

Section: Cellular Force Measurementmentioning

confidence: 99%

In Vitro Measurements of Cellular Forces and their Importance in the Lung—From the Sub- to the Multicellular Scale

Kolb

Schundner

Frick

et al. 2021

Life

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Throughout life, the body is subjected to various mechanical forces on the organ, tissue, and cellular level. Mechanical stimuli are essential for organ development and function. One organ whose function depends on the tightly connected interplay between mechanical cell properties, biochemical signaling, and external forces is the lung. However, altered mechanical properties or excessive mechanical forces can also drive the onset and progression of severe pulmonary diseases. Characterizing the mechanical properties and forces that affect cell and tissue function is therefore necessary for understanding physiological and pathophysiological mechanisms. In recent years, multiple methods have been developed for cellular force measurements at multiple length scales, from subcellular forces to measuring the collective behavior of heterogeneous cellular networks. In this short review, we give a brief overview of the mechanical forces at play on the cellular level in the lung. We then focus on the technological aspects of measuring cellular forces at many length scales. We describe tools with a subcellular resolution and elaborate measurement techniques for collective multicellular units. Many of the technologies described are by no means restricted to lung research and have already been applied successfully to cells from various other tissues. However, integrating the knowledge gained from these multi-scale measurements in a unifying framework is still a major future challenge.

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“…The 3D organization of keratin networks differs profoundly between different epithelia: for example, the simple epithelial cells lining the intestine contain a well-defined lateral and apical network, keratinocytes contain a dense 3D network encompassing the entire cytoplasm, and hepatocytes have a prominent submembraneous network ( Schwarz et al, 2015 ; Strnad et al, 2008 ). Network organization is furthermore affected by mechanical stress ( Beriault et al, 2012 ; Felder et al, 2008 ; Fois et al, 2013 ; Karsch et al, 2020 ; Lutz et al, 2020 ; Quinlan et al, 2017 ) and differentiation-status ( Coch and Leube, 2016 ; Iwatsuki and Suda, 2010 ; Leube et al, 2017 ; Quinlan et al, 2017 ; Tateishi et al, 2017 ). In addition, individual proteins such as Ndel1 and plectin have been shown to regulate keratin network organization ( Kim et al, 2021 ; Moch et al, 2016 ; Osmanagic-Myers et al, 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Quantitative mapping of keratin networks in 3D

Windoffer

Schwarz

Yoon

et al. 2022

eLife

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Mechanobiology requires precise quantitative information on processes taking place in specific 3D microenvironments. Connecting the abundance of microscopical, molecular, biochemical and cell mechanical data with defined topologies has turned out to be extremely difficult. Establishing such structural and functional 3D maps needed for biophysical modeling is a particular challenge for the cytoskeleton, which consists of long and interwoven filamentous polymers coordinating subcellular processes and interactions of cells with their environment. To date, useful tools are available for the segmentation and modeling of actin filaments and microtubules but comprehensive tools for the mapping of intermediate filament organization are still lacking. In this work, we describe a workflow to model and examine the complete 3D arrangement of the keratin intermediate filament cytoskeleton in canine, murine and human epithelial cells both, in vitro and in vivo. Numerical models are derived from confocal Airyscan high resolution 3D imaging of fluorescence-tagged keratin filaments. They are interrogated and annotated at different length scales using different modes of visualization including immersive virtual reality. In this way, information is provided on network organization at the subcellular level including mesh arrangement, density and isotropic configuration as well as details on filament morphology such as bundling, curvature and orientation. We show that the comparison of these parameters helps to identify, in quantitative terms, similarities and differences of keratin network organization in epithelial cell types defining subcellular domains, notably basal, apical, lateral and perinuclear systems. The described approach and the presented data are pivotal for generating mechanobiological models that can be experimentally tested.

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Acute effects of cell stretch on keratin filaments in A549 lung cells

Cited by 5 publications

References 57 publications

Plectin-mediated cytoskeletal crosstalk controls cell tension and cohesion in epithelial sheets

Plectin-mediated cytoskeletal crosstalk controls cell tension and cohesion in epithelial sheets

In Vitro Measurements of Cellular Forces and their Importance in the Lung—From the Sub- to the Multicellular Scale

Quantitative mapping of keratin networks in 3D

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