Effects of exogenous ganglioside GM1 on different stages of cell spreading studied by directly quantifying spreading rate

Huang, Jie; Shao, Wenxiang; Wu, Li; Yang, Wen Bin; Chen, Yong

doi:10.3109/15419061.2012.749245

Cited by 3 publications

(2 citation statements)

References 23 publications

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“…In presence of both integrin specific and unspecific ligands (fibronectin and vitronectin) the spreading curve became more linear and cells continued to spread radially even after 20 min from initial attachment. These data are consistent with other studies, as it was shown that serum promotes spreading but not initial attachment of 3T3, chicken embryonal fibroblasts and human skin fibroblasts (Edwards et al, 1993; Tucker et al, 1981; Verger and Imbenotte, 1990), and serum conditioning is not efficient for spreading of CHO and HUVEC cells (Edwards et al, 1993; Huang et al, 2012).…”

Section: Discussionsupporting

confidence: 92%

Dynamic microtubules drive fibroblast spreading

et al. 2018

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When cells with a mesenchymal type of motility come into contact with an adhesive substrate they adhere and start spreading by the formation of lamellipodia. Using a label-free approach and virtual synchronization approach we analyzed spreading in fibroblasts and cancer cells. In all cell lines spreading is a non-linear process undergoing isotropic or anisotropic modes with first fast (5–20 min) and then slow (30–120 min) phases. In the first 10 min cell area increases 2–4 times, while the absolute rate of initial spreading decreases 2–8 times. Fast spreading depends on actin polymerization and dynamic microtubules. Inhibition of microtubule growth was sufficient for a slowdown of initial spreading. Inhibition of myosin II in the presence of stable microtubules restored fast spreading. Inhibition of actin polymerization or complete depolymerization of microtubules slowed down fast spreading. However, in these cases inhibition of myosin II only partially restored spreading kinetics. We conclude that rapid growth of microtubules towards cell margins at the first stage of cell spreading temporarily inhibits phosphorylation of myosin II and is essential for the fast isotropic spreading. Comparison of the fibroblasts with cancer cells shows that fast spreading in different cell types shares similar kinetics and mechanisms, and strongly depends on dynamic microtubules.

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Section: Discussionsupporting

confidence: 92%

Dynamic microtubules drive fibroblast spreading

et al. 2018

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“…14,15) Briefly, HUVECs at 70% confluence were treated with or without 10 mM MβCD for 30 min at 37°C. The cells were then fixed by 4% paraformaldehyde, rinsed with PBS, and imaged in PBS by the confocal microscope.…”

Section: Methodsmentioning

confidence: 99%

Methyl-β-Cyclodextrin Impairs the Monocyte-Adhering Ability of Endothelial Cells by Down-Regulating Adhesion Molecules and Caveolae and Reorganizing the Actin Cytoskeleton

Zhou

et al. 2016

Biological & Pharmaceutical Bulletin

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Due to its powerful ability to deplete cholesterol from the plasma membrane of cells, methyl-β-cyclodextrin (MβCD) has been widely used as a putative research tool in cell biology. Recently, recruiting MβCD as an effective drug (e.g., antitumor drugs) has been developed. However, it remains unclear whether MβCD, when it enters the blood circulation as a drug, influences the functions of the endothelium, e.g., the adhesion of leukocytes to the endothelium. In this study, we found that MβCD can impair the adhesion of monocytes to the monolayer of endothelial cells by lowering the cell-surface adhesive force and expression of adhesion molecules and caveolae-related molecules on/in endothelial cells, and reorganizing the actin cytoskeleton of endothelial cells. The data imply that MβCD, when recruited as a drug, potentially helps to inhibit inflammation or initiation/progression of atherosclerosis since its important early step is the adhesion of circulating leukocytes (e.g., monocytes) to the endothelium.Key words methyl-β-cyclodextrin; monocyte; endothelial cell; adhesion molecule; caveola; actin cytoskeleton Beta-cyclodextrin (β-CD) is a cyclic oligosaccharide consisting of seven α-(1,4)-linked glucopyranose subunits. Due to its relatively low water solubility (approximately 2% weight by weight), 1) various derivatives of β-CD have been developed to significantly improve the water solubility, among which methyl-β-cyclodextrin (MβCD) is the one studied most intensively. MβCD is well known as a putative powerful cholesterol-depleting agent and commonly used to deplete plasma membrane cholesterol from cultured mammalian cells. 2-4)MβCD-cholesterol complexes also can efficiently replenish cells with cholesterol as cholesterol donors. 3,4) Moreover, due to the structure of a lipophilic internal cavity surrounded by hydrophilic outer surface, MβCD was applied as a drug delivery system. 5) Recently, it was found that MβCD per se potentially has antitumor effects and has the possibility of being an effective antitumor drug.6-11) However, it remains uncertain whether MβCD, as an anticancer drug, influences the functions of the endothelium, e.g. the adhesion of circulating leukocytes (e.g. monocytes) onto the endothelium which is an important step during inflammation or initiation/progression of atherosclerosis.In this study, the effect of MβCD on the adhesion of monocytes onto the monolayer of endothelial cells was first evaluated. Then, the effects of MβCD on cell-surface adhesive force properties, expression/distribution of adhesion molecules and caveolae-related molecules, and actin cytoskeleton of endothelial cells were investigated to elucidate the potential underlying mechanisms. MATERIALS AND METHODSCell Culture Human umbilical vein endothelial cells (HUVECs) were purchased from Xiangya Central Experiment Laboratory (Hunan, China) and cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco, U.S.A.) supplemented with 10% (w/v) fetal calf serum (Hyclone, South Logan, UT, U.S.A), 100 U/mL penicillin, and 100 µg/m...

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AFM of the Ultrastructural and Mechanical Properties of Lipid-Raft-Disrupted and/or Cold-Treated Endothelial Cells

Huang

et al. 2014

J Membrane Biol

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The nonionic detergent extraction at 4 °C and the cholesterol-depletion-induced lipid raft disruption are the two widely used experimental strategies for lipid raft research. However, the effects of raft disruption and/or cold treatment on the ultrastructural and mechanical properties of cells are still unclear. Here, we evaluated the effects of raft disruption and/or cold (4 °C) treatment on these properties of living human umbilical vein endothelial cells (HUVECs). At first, the cholesterol-depletion-induced raft disruption was visualized by confocal microscopy and atomic force microscopy (AFM) in combination with fluorescent quantum dots. Next, the cold-induced cell contraction and the formation of end-branched filopodia were observed by confocal microscopy and AFM. Then, the cell-surface ultrastructures were imaged by AFM, and the data showed that raft disruption and cold treatment induced opposite effects on cell-surface roughness (a significant decrease and a significant increase, respectively). Moreover, the cell-surface mechanical properties (stiffness and adhesion force) of raft-disrupted- and/or cold-treated HUVECs were measured by the force measurement function of AFM. We found that raft disruption and cold treatment induced parallel effects on cell stiffness (increase) or adhesion force (decrease) and that the combination of the two treatments caused dramatically strengthened effects. Finally, raft disruption was found to significantly impair cell migration as previously reported, whereas temporary cold treatment only caused a slight but nonsignificant decrease in cell migration performed at physiological temperature. Although the mechanisms for causing these results might be complicated and more in-depth studies will be needed, our data may provide important information for better understanding the effects of raft disruption or cold treatment on cells and the two strategies for lipid raft research.

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Effects of exogenous ganglioside GM1 on different stages of cell spreading studied by directly quantifying spreading rate

Cited by 3 publications

References 23 publications

Dynamic microtubules drive fibroblast spreading

Dynamic microtubules drive fibroblast spreading

Methyl-β-Cyclodextrin Impairs the Monocyte-Adhering Ability of Endothelial Cells by Down-Regulating Adhesion Molecules and Caveolae and Reorganizing the Actin Cytoskeleton

AFM of the Ultrastructural and Mechanical Properties of Lipid-Raft-Disrupted and/or Cold-Treated Endothelial Cells

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