scholarly journals Force-induced recruitment of cten along keratin network in epithelial cells

2019 ◽  
Vol 116 (40) ◽  
pp. 19799-19801 ◽  
Author(s):  
Joleen S. Cheah ◽  
Kyle A. Jacobs ◽  
Volkmar Heinrich ◽  
Su Hao Lo ◽  
Soichiro Yamada

The cytoskeleton provides structural integrity to cells and serves as a key component in mechanotransduction. Tensins are thought to provide a force-bearing linkage between integrins and the actin cytoskeleton; yet, direct evidence of tensin’s role in mechanotransduction is lacking. We here report that local force application to epithelial cells using a micrometer-sized needle leads to rapid accumulation of cten (tensin 4), but not tensin 1, along a fibrous intracellular network. Surprisingly, cten-positive fibers are not actin fibers; instead, these fibers are keratin intermediate filaments. The dissociation of cten from tension-free keratin fibers depends on the duration of cell stretch, demonstrating that the external force favors maturation of cten−keratin network interactions over time and that keratin fibers retain remarkable structural memory of a cell’s force-bearing state. These results establish the keratin network as an integral part of force-sensing elements recruiting distinct proteins like cten and suggest the existence of a mechanotransduction pathway via keratin network.

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2058
Author(s):  
Nicole Reisinger ◽  
Dominik Wendner ◽  
Nora Schauerhuber ◽  
Elisabeth Mayer

Endotoxins play a crucial role in ruminant health due to their deleterious effects on animal health. The study aimed to evaluate whether LPS and LTA can induce an inflammatory response in rumen epithelial cells. For this purpose, epithelial cells isolated from rumen tissue (RECs) were stimulated with LPS and LTA for 1, 2, 4, and 24 h. Thereafter, the expression of selected genes of the LPS and LTA pathway and inflammatory response were evaluated. Furthermore, it was assessed whether LPS affects inflammatory response and structural integrity of claw explants. Therefore, claw explants were incubated with LPS for 4 h to assess the expression of selected genes and for 24 h to evaluate tissue integrity via separation force. LPS strongly affected the expression of genes related to inflammation (NFkB, TNF-α, IL1B, IL6, CXCL8, MMP9) in RECs. LTA induced a delayed and weaker inflammatory response than LPS. In claw explants, LPS affected tissue integrity, as there was a concentration-dependent decrease of separation force. Incubation time had a strong effect on inflammatory genes in claw explants. Our data suggest that endotoxins can induce a local inflammatory response in the rumen epithelium. Furthermore, translocation of LPS might negatively impact claw health.


2018 ◽  
Vol 315 (4) ◽  
pp. G433-G442 ◽  
Author(s):  
Kayte A. Jenkin ◽  
Peijian He ◽  
C. Chris Yun

Lysophosphatidic acid (LPA) is a bioactive lipid molecule, which regulates a broad range of pathophysiological processes. Recent studies have demonstrated that LPA modulates electrolyte flux in the intestine, and its potential as an antidiarrheal agent has been suggested. Of six LPA receptors, LPA5 is highly expressed in the intestine. Recent studies by our group have demonstrated activation of Na+/H+ exchanger 3 (NHE3) by LPA5. However, much of what has been elucidated was achieved using colonic cell lines that were transfected to express LPA5. In the current study, we engineered a mouse that lacks LPA5 in intestinal epithelial cells, Lpar5ΔIEC, and investigated the role of LPA5 in NHE3 regulation and fluid absorption in vivo. The intestine of Lpar5ΔIEC mice appeared morphologically normal, and the stool frequency and fecal water content were unchanged compared with wild-type mice. Basal rates of NHE3 activity and fluid absorption and total NHE3 expression were not changed in Lpar5ΔIEC mice. However, LPA did not activate NHE3 activity or fluid absorption in Lpar5ΔIEC mice, providing direct evidence for the regulatory role of LPA5. NHE3 activation involves trafficking of NHE3 from the terminal web to microvilli, and this mobilization of NHE3 by LPA was abolished in Lpar5ΔIEC mice. Dysregulation of NHE3 was specific to LPA, and insulin and cholera toxin were able to stimulate and inhibit NHE3, respectively, in both wild-type and Lpar5ΔIEC mice. The current study for the first time demonstrates the necessity of LPA5 in LPA-mediated stimulation of NHE3 in vivo. NEW & NOTEWORTHY This study is the first to assess the role of LPA5 in NHE3 regulation and fluid absorption in vivo using a mouse that lacks LPA5 in intestinal epithelial cells, Lpar5ΔIEC. Basal rates of NHE3 activity and fluid absorption, and total NHE3 expression were not changed in Lpar5ΔIEC mice. However, LPA did not activate NHE3 activity or fluid absorption in Lpar5ΔIEC mice, providing direct evidence for the regulatory role of LPA5.


2010 ◽  
Vol 138 (5) ◽  
pp. S-589-S-590
Author(s):  
Hongguang Liu ◽  
Amika Singla ◽  
Mei Ao ◽  
Ravinder K. Gill ◽  
Jayashree Venkatasubramanian ◽  
...  

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Kristin M Sherrard ◽  
Maureen Cetera ◽  
Sally Horne-Badovinac

Stress fibers (SFs) are actomyosin bundles commonly found in individually migrating cells in culture. However, whether and how cells use SFs to migrate in vivo or collectively is largely unknown. Studying the collective migration of the follicular epithelial cells in Drosophila, we found that the SFs in these cells show a novel treadmilling behavior that allows them to persist as the cells migrate over multiple cell lengths. Treadmilling SFs grow at their fronts by adding new integrin-based adhesions and actomyosin segments over time. This causes the SFs to have many internal adhesions along their lengths, instead of adhesions only at the ends. The front-forming adhesions remain stationary relative to the substrate and typically disassemble as the cell rear approaches. By contrast, a different type of adhesion forms at the SF’s terminus that slides with the cell’s trailing edge as the actomyosin ahead of it shortens. We further show that SF treadmilling depends on cell movement and identify a developmental switch in the formins that mediate SF assembly, with Dishevelled-associated activator of morphogenesis acting during migratory stages and Diaphanous acting during postmigratory stages. We propose that treadmilling SFs keep each cell on a linear trajectory, thereby promoting the collective motility required for epithelial migration.


Nanomedicine ◽  
2021 ◽  
Author(s):  
Samuel Bonnet ◽  
Rana Elfatairi ◽  
Florence Franconi ◽  
Emilie Roger ◽  
Samuel Legeay

To understand how nanoparticles (NPs) interact with biological barriers and to ensure they maintain their integrity over time, it is crucial to study their in vivo pharmacokinetic (PK) profiles. Many methods of tracking have been used to describe the in vivo fate of NPs and to evaluate their PKs and structural integrity. However, they do not deliver the same level of information and this may cause misinterpretations. Here, the authors review and discuss the different methods for in vivo tracking of organic NPs. Among them, Förster resonance energy transfer (FRET) presents great potential to track NPs' integrity. However, FRET still requires validated methods to extract and quantify NPs in biological fluids and tissues.


1990 ◽  
Vol 68 (9) ◽  
pp. 1839-1856 ◽  
Author(s):  
S. J. Hall ◽  
J. H. Youson

Light microscopic autoradiography and quantitative tissue analyses were used to examine the distribution of radioiron in larvae (ammocoetes) of the lamprey Petromyzon marinus L. over time following the injection of 55ferrous citrate (55Fe). It was found that a single intraperitoneal injection of 55Fe is an efficient method of introducing iron into ammocoetes, and radioiron retention within the body remained high throughout the 28-d experiment. Immediately after radioiron administration the isotope was transported in the serum fraction of the blood to sites of iron storage and elimination. The liver is the most important target organ for iron in ammocoetes, and radioiron acquired from the alimentary canal, skin, carcass, and blood contributed to the initial increase in this organ. Data indicated only temporary storage in the liver; between 3 and 16 d, radioiron was redistributed by the blood to the skin, kidney, alimentary canal, and carcass. Biliary transport of radioiron from the liver to the lumen of the anterior intestine may also explain the lowering of iron concentrations in the liver and the rise in levels in the alimentary canal during this interval. The redeposition of 55Fe in the liver at 16 d may signify longer-term storage of the metal in this organ. There is movement of radioiron in and out of the carcass tissues over time, and most of the radioactivity is localized in the adipose tissue. The epithelial cells of the proximal tubules of the kidney absorb and concentrate the radioiron presumably present in the glomerular filtrate. This concentration probably reflects both the absorption of iron bound to ferritin and a mechanism necessary for maintaining the high serum iron levels of ammocoetes. The release of mucus from iron-laden mucous cells of the skin and exfoliation of radioiron-laden epithelial cells in specific regions of the posterior intestine are possible routes for iron elimination. However, percent incorporation of the injected radioactivity was still high at the end of the experiment, indicating that iron excretion in ammocoetes is relatively low or that it is a slow process. These data support the notion that lampreys provide a useful animal model for the study of iron metabolism in vertebrates.


2012 ◽  
Vol 53 (1) ◽  
pp. 37-43 ◽  
Author(s):  
Christopher J. Harmer ◽  
James A. Triccas ◽  
Honghua Hu ◽  
Barbara Rose ◽  
Peter Bye ◽  
...  

2000 ◽  
Vol 149 (3) ◽  
pp. 647-656 ◽  
Author(s):  
Ronald R. Dubreuil ◽  
Ping Wang ◽  
Steve Dahl ◽  
John Lee ◽  
Lawrence S.B. Goldstein

Spectrin has been proposed to function as a sorting machine that concentrates interacting proteins such as the Na,K ATPase within specialized plasma membrane domains of polarized cells. However, little direct evidence to support this model has been obtained. Here we used a genetic approach to directly test the requirement for the β subunit of the αβ spectrin molecule in morphogenesis and function of epithelial cells in Drosophila. β Spectrin mutations were lethal during late embryonic/early larval development and they produced subtle defects in midgut morphology and stomach acid secretion. The polarized distributions of αβH spectrin and ankyrin were not significantly altered in β spectrin mutants, indicating that the two isoforms of Drosophila spectrin assemble independently of one another, and that ankyrin is upstream of αβ spectrin in the spectrin assembly pathway. In contrast, β spectrin mutations had a striking effect on the basolateral accumulation of the Na,K ATPase. The results establish a role for β spectrin in determining the subcellular distribution of the Na,K ATPase and, unexpectedly, this role is independent of α spectrin.


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