scholarly journals Sustained oscillations of epithelial cell sheets

2018 ◽  
Author(s):  
Grégoire Peyret ◽  
Romain Mueller ◽  
Joseph d’Alessandro ◽  
Simon Begnaud ◽  
Philippe Marcq ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Large Scale ◽  
Morphological Changes ◽  
Computational Modelling ◽  
Cell Junctions ◽  
Force Transmission ◽  
Cell Sheets ◽  
Wide Range ◽  
Collective Oscillations

AbstractMorphological changes during development, tissue repair, and disease largely rely on coordinated cell movements and are controlled by the tissue environment. Epithelial cell sheets are often subjected to large scale deformation during tissue formation. The active mechanical environment in which epithelial cells operate have the ability to promote collective oscillations, but how these cellular movements are generated and relate to collective migration remains unclear. Here, combining in vitro experiments and computational modelling we describe a novel mode of collective oscillations in confined epithelial tissues where the oscillatory motion is the dominant contribution to the cellular movements. We show that epithelial cells exhibit large-scale coherent oscillations when constrained within micro-patterns of varying shapes and sizes, and that their period and amplitude are set by the smallest confinement dimension. Using molecular perturbations, we then demonstrate that force transmission at cell-cell junctions and its coupling to cell polarity are pivotal for the generation of these collective movements. We find that the resulting tissue deformations are sufficient to trigger mechanotransduction within cells, potentially affecting a wide range of cellular processes.

scholarly journals Keratin intermediate filaments: intermediaries of epithelial cell migration

2019 ◽  
Vol 63 (5) ◽  
pp. 521-533 ◽  
Author(s):  
Sungjun Yoon ◽  
Rudolf E. Leube
Keyword(s):  
Cell Migration ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Biomechanical Properties ◽  
Biochemical Properties ◽  
Cell Junctions ◽  
Epithelial Tissue ◽  
Post Translational Modification ◽  
Cell Sheets ◽  
Epithelial Cell Migration

Abstract Migration of epithelial cells is fundamental to multiple developmental processes, epithelial tissue morphogenesis and maintenance, wound healing and metastasis. While migrating epithelial cells utilize the basic acto-myosin based machinery as do other non-epithelial cells, they are distinguished by their copious keratin intermediate filament (KF) cytoskeleton, which comprises differentially expressed members of two large multigene families and presents highly complex patterns of post-translational modification. We will discuss how the unique mechanophysical and biochemical properties conferred by the different keratin isotypes and their modifications serve as finely tunable modulators of epithelial cell migration. We will furthermore argue that KFs together with their associated desmosomal cell–cell junctions and hemidesmosomal cell–extracellular matrix (ECM) adhesions serve as important counterbalances to the contractile acto-myosin apparatus either allowing and optimizing directed cell migration or preventing it. The differential keratin expression in leaders and followers of collectively migrating epithelial cell sheets provides a compelling example of isotype-specific keratin functions. Taken together, we conclude that the expression levels and specific combination of keratins impinge on cell migration by conferring biomechanical properties on any given epithelial cell affecting cytoplasmic viscoelasticity and adhesion to neighboring cells and the ECM.

scholarly journals ROCK inhibitor combined with Ca2+ controls the myosin II activation and optimizes human nasal epithelial cell sheets

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yoshiyuki Kasai ◽  
Tsunetaro Morino ◽  
Eri Mori ◽  
Kazuhisa Yamamoto ◽  
Hiromi Kojima
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Sheet ◽  
Cell Junctions ◽  
Rock Inhibitor ◽  
Cell Sheets ◽  
Cultured Epithelial Cells ◽  
Rock Inhibition ◽  
Mlc Phosphorylation ◽  
Cell Cell

Abstract The proliferation and differentiation of cultured epithelial cells may be modified by Rho-associated kinase (ROCK) inhibition and extracellular Ca2+ concentration. However, it was not known whether a combination would influence the behavior of cultured epithelial cells through changes in the phosphorylation of non-muscle myosin light chain II (MLC). Here we show that the combination of ROCK inhibition with Ca2+ elevation regulated the phosphorylation of MLC and improved both cell expansion and cell–cell adhesion during the culture of human nasal mucosal epithelial cell sheets. During explant culture, Ca2+ enhanced the adhesion of nasal mucosal tissue, while ROCK inhibition downregulated MLC phosphorylation and promoted cell proliferation. During cell sheet culture, an elevation of extracellular Ca2+ promoted MLC phosphorylation and formation of cell–cell junctions, allowing the harvesting of cell sheets without collapse. Moreover, an in vitro grafting assay revealed that ROCK inhibition increased the expansion of cell sheets three-fold (an effect maintained when Ca2+ was also elevated), implying better wound healing potential. We suggest that combining ROCK inhibition with elevation of Ca2+ could facilitate the fabrication of many types of cell graft.

scholarly journals An asymmetric junctional mechanoresponse coordinates mitotic rounding with epithelial integrity

2021 ◽  
Vol 220 (5) ◽  
Author(s):  
Jooske L. Monster ◽  
Lisa Donker ◽  
Marjolein J. Vliem ◽  
Zaw Win ◽  
Helen K. Matthews ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Morphological Changes ◽  
Mitotic Cell ◽  
Cell Junctions ◽  
Actin Binding ◽  
Epithelial Barrier ◽  
Epithelial Integrity ◽  
Tensile Forces ◽  
Shape Changes ◽  
Mitotic Cells

Epithelia are continuously self-renewed, but how epithelial integrity is maintained during the morphological changes that cells undergo in mitosis is not well understood. Here, we show that as epithelial cells round up when they enter mitosis, they exert tensile forces on neighboring cells. We find that mitotic cell–cell junctions withstand these tensile forces through the mechanosensitive recruitment of the actin-binding protein vinculin to cadherin-based adhesions. Surprisingly, vinculin that is recruited to mitotic junctions originates selectively from the neighbors of mitotic cells, resulting in an asymmetric composition of cadherin junctions. Inhibition of junctional vinculin recruitment in neighbors of mitotic cells results in junctional breakage and weakened epithelial barrier. Conversely, the absence of vinculin from the cadherin complex in mitotic cells is necessary to successfully undergo mitotic rounding. Our data thus identify an asymmetric mechanoresponse at cadherin adhesions during mitosis, which is essential to maintain epithelial integrity while at the same time enable the shape changes of mitotic cells.

scholarly journals An intact keratin network is crucial for mechanical integrity and barrier function in keratinocyte cell sheets

2019 ◽  
Author(s):  
Susanne Karsch ◽  
Fanny Büchau ◽  
Thomas M. Magin ◽  
Andreas Janshoff
Keyword(s):  
Barrier Function ◽  
Time Course ◽  
Mechanical Response ◽  
Adhesion Force ◽  
Cell Junctions ◽  
Force Transmission ◽  
Cell Sheets ◽  
Mechanical Integrity ◽  
Barrier Formation ◽  
Mechanical Homeostasis

AbstractThe isotype-specific composition of the keratin cytoskeleton is important for strong adhesion, force resilience, and barrier function of the epidermis. However, the mechanisms by which keratins regulate these functions are still incompletely understood. In this study, the role and significance of the keratin network for mechanical integrity, force transmission, and barrier formation were analyzed in murine keratinocytes. Following the time-course of single-cell wounding, wildtype (WT) cells slowly closed the gap in a collective fashion involving tightly connected neighboring cells. In contrast, the mechanical response of neighboring cells was compromised in keratin-deficient cells, causing an increased wound area initially and an inefficient overall wound closure. Furthermore, the loss of the keratin network led to impaired, fragmented cell-cell junctions and triggered a profound change in the overall cellular actomyosin architecture. Electrical cell-substrate impedance sensing of cell junctions revealed a dysfunctional barrier in knockout (Kty−/−) compared to WT cells. These findings demonstrate that Kty−/− cells display a novel phenotype characterized by loss of mechanocoupling and failure to form a functional barrier. Re-expression of K5/K14 rescued the barrier defect to a significant extent and reestablished the mechanocoupling with remaining discrepancies likely due to the low abundance of keratins in that setting. Our study reveals the major role of the keratin network for mechanical homeostasis and barrier functionality in keratinocyte layers.

scholarly journals The splicing regulators Esrp1 and Esrp2 direct an epithelial splicing program essential for mammalian development

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Thomas W Bebee ◽  
Juw Won Park ◽  
Katherine I Sheridan ◽  
Claude C Warzecha ◽  
Benjamin W Cieply ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Large Scale ◽  
Cleft Lip ◽  
Cleft Lip And Palate ◽  
Regulatory Protein ◽  
Cell Type ◽  
Cell Functions ◽  
Essential Components ◽  
Cell Type Specific

Tissue- and cell-type-specific regulators of alternative splicing (AS) are essential components of posttranscriptional gene regulation, necessary for normal cellular function, patterning, and development. Mice with ablation of Epithelial splicing regulatory protein (Esrp1) develop cleft lip and palate. Loss of both Esrp1 and its paralog Esrp2 results in widespread developmental defects with broad implications to human disease. Deletion of the Esrps in the epidermis revealed their requirement for establishing a proper skin barrier, a primary function of epithelial cells comprising the epidermis. We profiled the global Esrp-mediated splicing regulatory program in epidermis, which revealed large-scale programs of epithelial cell-type-specific splicing required for epithelial cell functions. These mice represent a valuable model for evaluating the essential role for AS in development and function of epithelial cells, which play essential roles in tissue homeostasis in numerous organs, and provide a genetic tool to evaluate important functional properties of epithelial-specific splice variants in vivo.

scholarly journals Lactobacillus Rhamnosus GR-1 Ameliorates Trueperella Pyogenes–Induced Barrier Dysfunction of Bovine Endometrial Epithelial Cells

2020 ◽  
Author(s):  
Ning Liu ◽  
Xuan Wu ◽  
Mengling Wang ◽  
Yanan Li ◽  
Le Xu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Morphological Changes ◽  
Lactobacillus Rhamnosus ◽  
Tunel Staining ◽  
Barrier Dysfunction ◽  
Trueperella Pyogenes ◽  
Effector Caspase ◽  
Wide Range ◽  
Endometrial Epithelial Cells ◽  
In Cells

Trueperella pyogenes is a common opportunistic pathogen which is one of the main causes of postpartum endometritis in dairy cows. As a substitute for antibiotics, the probiotic Lactobacillus rhamnosus GR-1 has been used in a wide range of clinical treatments. Our experiments were designed to establish a model of anti-damage which LGR-1 was used to protect bovine endometrial epithelial cells (BEECs) from inflammatory damage and cell destruction caused by T. pyogenes. Increased expression of NLRP3 inflammasomes and cytokines was observed following T. pyogenes challenge, but this increase was relieved by LGR-1 pretreatment. Immunofluorescence and Western blot analyses revealed that T. pyogenes infection also results in the damage of tight junction proteins in BEECs. The expression levels of Claudin-1, Occludin, and ZO-1 were decreased in cells only infected with T. pyogenes but not in cells pretreated with LGR-1. Moreover, the detection of the anti-apoptotic protein Bcl-2 and apoptotic proteins BAX, cytochrome c, as well as the activating effector caspase-3 revealed that T. pyogenes induced apoptosis of BEECs, which was also confirmed by DAPI staining to observe the morphological changes of the nuclei of cell apoptosis and by TUNEL staining to locate the cells undergoing apoptosis. Our data indicate that LGR-1 ameliorates the T. pyogenes–induced barrier dysfunction of BEECs and pre-application of LGR-1 could be an effective strategy for controlling T. pyogenes infection.

scholarly journals Biobanked human foreskin epithelial cell sheets reduce inflammation and promote wound healing in a nude mouse model

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Dongliang Zhang ◽  
Jialiang Shao ◽  
Jingming Zhuang ◽  
Shukui Zhou ◽  
Shuo Yin ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Wound Healing ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Sheet ◽  
Skin Defect ◽  
Cell Sheets ◽  
Vascular Regeneration ◽  
Middle Ear Cholesteatoma

Abstract Background Human epithelial cell sheets (ECSs) are used to clinically treat epithelial conditions such as burns, corneal blindness, middle ear cholesteatoma and vitiligo. As a widely used material in clinic, there is little information on the biobanking of ECSs and its repair effect after storage. Results Two methods for biobanking foreskin ECSs were compared in a short term (7 days): 4-degree storage and programmed cryopreservation. Cell sheet integrity, viability, apoptosis, immunogenicity, mechanical properties and function were evaluated. In vivo, ECSs were directly transplanted to skin defect models and histological examination was performed at 1 week postoperatively. We successfully extracted human foreskin-derived primary epithelial cells and fabricated them into ECSs. Compared with 4-degree storage, programmed cryopreservation preserved the ECS structural integrity, enhanced the mechanical properties, decreased HLA-I expression, and increased cell viability and survival. An increased proportion of melanocytes with proliferative capacity remained in the cryopreserved sheets, and the undifferentiated epithelial cells were comparable to those of the fresh sheets. In vivo, cryopreserved ECSs could reduce inflammatory cell infiltration and promote connective tissue remodeling, epithelial cell proliferation and vascular regeneration. Conclusions Programmed cryopreservation of ECSs was superior and more feasible than 4-degree storage and the cryopreserved ECSs achieved satisfying skin wound healing in vivo. We anticipate that the off-the-shelf ECSs could be quickly used, such as, to repair human epithelial defect in future. Graphical abstract

scholarly journals Measurement of junctional tension in epithelial cells at the onset of primitive streak formation in the chick embryo via non-destructive optical manipulation

2018 ◽  
Author(s):  
Valentina Ferro ◽  
Manli Chuai ◽  
David McGloin ◽  
Cornelis Weijer
Keyword(s):  
Epithelial Cells ◽  
Cell Shape ◽  
Large Scale ◽  
Cell Sheet ◽  
Primitive Streak ◽  
Cell Junctions ◽  
Optical Manipulation ◽  
Cell Shape Changes ◽  
Non Destructive ◽  
Shape Changes

Oriented cell intercalations and cell shape changes are key determinants of large-scale epithelial cell sheet deformations occurring during gastrulation in many organisms. In several cases directional intercalation and cell shape changes have been shown to be associated with a planar cell polarity in the organisation of the actin-myosin cytoskeleton of epithelial cells. This polarised cytoskeletal organisation has been postulated to reflect the directional tension necessary to drive and orient directional cell intercalations. We have now further characterised and applied a recently introduced non-destructive optical manipulation technique to measure the tension in individual cell junctions in the epiblast of chick embryos in the early stages of primitive streak formation. We have measured junctional tension as a function of position and orientation. Junctional tension of mesendoderm cells, the tissue that drives the formation of the streak, is higher than tension of junctions of cells in other parts of the epiblast. Furthermore, in the mesendoderm junctional tension is higher in the direction of intercalation. The data are fitted best with a Maxwell model and we find that both junctional tension and relaxation time are dependent on myosin activity.

Inhibition of intestinal epithelial cell renewal and migration induced by starvation

1963 ◽  
Vol 205 (5) ◽  
pp. 868-872 ◽  
Author(s):  
H. Oliver Brown ◽  
Milton L. Levine ◽  
Martin Lipkin
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Intestinal Epithelium ◽  
Intestinal Epithelial Cell ◽  
Morphological Changes ◽  
Extraction Procedure ◽  
Chemical Extraction ◽  
Cell Renewal ◽  
Intestinal Epithelial ◽  
And Migration

Following starvation in mice, the rate of cell renewal of intestinal epithelium was measured. Injection of thymidine-H3 and microautoradiography, and thymidine-C14 and chemical extraction procedure were utilized. Cell renewal was reduced to about one-half the normal rate following extreme starvation. Morphological changes and impaired differentiation of the epithelial cells accompanied this change. The rate of migration of the epithelial cells to the villus tips was also reduced.

scholarly journals Significant Interference with Porcine Epidemic Diarrhea Virus Pandemic and Classical Strain Replication in Small-Intestine Epithelial Cells Using an shRNA Expression Vector

Vaccines ◽  
2019 ◽  
Vol 7 (4) ◽  
pp. 173
Author(s):  
Da Shi ◽  
Xiaobo Wang ◽  
Hongyan Shi ◽  
Jiyu Zhang ◽  
Yuru Han ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Porcine Epidemic Diarrhea Virus ◽  
Large Scale ◽  
N Gene ◽  
Shrna Expression ◽  
Diarrhea Virus ◽  
Shrna Expression Vector ◽  
Wide Range ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea (PED) re-emerged in China in 2010 and is now widespread. Evidence indicates that highly virulent porcine epidemic diarrhea virus (PEDV) strains belonging to genotype G2 caused a large-scale outbreak of diarrhea. Currently, vaccines derived from PEDV classical strains do not effectively prevent infection by virulent PEDV strains, and no specific drug is available to treat the disease. RNA interference (RNAi) is a novel and effective way to cure a wide range of viruses. We constructed three short hairpin RNA (shRNA)-expressing plasmids (shR-N307, shR-N463, and shR-N1071) directed against nucleocapsid (N) and determined their antiviral activities in intestine epithelial cells infected with a classical CV777 strain and LNCT2. We verified that shR-N307, shR-N463, and shR-N1071 effectively inhibited the expression of the transfected N gene in vitro, comparable to the control shRNA. We further demonstrated the shRNAs markedly reduced PEDV CV777 and LNCT2 replication upon downregulation of N production. Therefore, this study provides a new strategy for the design of antiviral methods against coronaviruses by targeting their processivity factors.

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