Physiologically induced restructuring of focal adhesions causes mobilization of vinculin by a vesicular endocytic recycling pathway

Márquez, Marı́a Gabriela; Brandán, Yamila Romina; Guaytima, Edith del Valle; Paván, Carlos Humberto; Favale, Nicolás Octavio; Sterín-Speziale, Norma

doi:10.1016/j.bbamcr.2014.09.014

Cited by 6 publications

(4 citation statements)

References 33 publications

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“…A proteomic analysis demonstrated the presence of Rab5 and EEA1 within isolated integrin adhesion complexes (27). Furthermore, vinculin, another scaffolding protein has been found to be associated with endosomal vesicles and clathrin-mediated endocytosis (28,29). This is consistent with a study that demonstrated a colocalization between FA and clathrin adaptors (Dab2, AP-1 and numb), which may be found in the plasma membrane and early endosomes (17).…”

Section: Discussionsupporting

confidence: 84%

Roles of Endocytic Processes and Early Endosomes on Focal Adhesion Dynamics in MDA-MB-231 Cells

2021

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Background: Focal adhesion (FA) play a critical role in many biological processes which include cell survival and cell migration. They serve as cellular anchor, allowing cells to stay attached to the extracellular matrix (ECM), and can also regulate cellular transduction. Previously, it has been suggested that vesicles such as endosomes could interact directly with FA or be implicated in their turnover. In this study, we investigated whether there is a relationship between FA and the early endocytic machinery in MDA-MB-231 cells. Methods: In this study, cell culture, transfection, time laps confocal microscopies, immunocytochemistry, western blotting, Cell fractionation and immunoprecipitation techniques were performed. Results: Cells acutely treated with Dynasore, an inhibitor of dynamin, or with Pitstop 2, an inhibitor of clathryn-dependent endocytosis showed a reduction in the expression of early endosome biomarkers such as Rab5 and EEA1. Additionally, cells treated with these endocytic inhibitors exhibited an increase number and size of FA, as well as an increase FA turnover duration. This data was consistent with the reduction of the speed of cell migration. We demonstrated that Rab5-and EEA1-positive early endosomes were found to be colocalized with internalized FA. Conclusions:The present study suggests that there is a link between FA and early endosome markers, which indicates that the early endosomes may be involved in FA dynamics.

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Section: Discussionsupporting

confidence: 84%

Roles of Endocytic Processes and Early Endosomes on Focal Adhesion Dynamics in MDA-MB-231 Cells

2021

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“…79,80 Furthermore, different cell types react differently to biomaterials. [81][82][83] Therefore, it is difficult to draw a conclusive explanation for the different cell behaviour of MC3T3 cells when cultured on pTMSPMA/SiO 2 and I100 disks. The combination of a more hydrophilic, negatively charged surface and the inherent chemistry of pTMSPMA is likely to contribute to the improved cell attachment and spreading.…”

Section: Cell Viabilitymentioning

confidence: 99%

Ductile silica/methacrylate hybrids for bone regeneration

Maçon

Chung

et al. 2016

J. Mater. Chem. B

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BioglassR was the first synthetic material capable of bonding with bone without fibrous encapsulation, and fulfils some of the criteria of an ideal synthetic bone graft. However, it is brittle and toughness is required. Here, we investigated hybrids consisting of co-networks of high crosslinking density polymethacrylate and silica (class II hybrid) as a potential new generation of scaffold materials. Poly(3-(methoxysilyl)propyl methacrylate) (pTMSPMA) and tetraethyl orthosilicate (TEOS) were used as sol-gel precursors and hybrids were synthesised with different inorganic to organic ratios (I h ). The hybrids were nanoporous, with a modal pore diameter of 1 nm. At I h = 50 %, the release of silica was controlled by varying the molecular weight of pTMSPMA while retaining a specific surface area above 100 m 2 g −1 . Strain to failure increased to 14.2 %, for I h = 50 % using a polymer of 30 kDa, compared to 4.5 % for pure glass. The modulus of toughness (U T ) increased from 0.73 (pure glass) to 2.64 GPa. Although, the hybrid synthesised in this report did not contain calcium, pTMSPMA/SiO 2 hybrid was found to nucleate bone-like mineral on its surface after 1 week of immersion in simulated body fluid (SBF), whereas pure silica sol-gel glass did not. This increase in apatite forming ability was due to the ion-dipole complexation of calcium with the ester moieties of the polymer that were exposed after release of soluble silica from TEOS. No adverse cytotoxicity for MC3T3-E1 osteoblast-like cells was detected and improved cell attachment was observed, compared to a pure silica gel. pTMSPMA/SiO 2 hybrids have potential for the regeneration of hard tissue as they overcome the major drawbacks of pure inorganic substrates while retaining cell attachment.Bone is the second most transplanted tissue after blood and musculoskeletal pathology treatments represent £10 billion per year in the U.K. alone. 1,2 Effective alternatives must be found to the current methods of skeletal defect treatments to avoid postoperation infection or revision, which can cost up to £70,000 per patient. 3 One attractive approach is to design biodegradable implants that could actively promote the bone growth and tissue remodelling by stimulating cellular activities whilst providing a mechanical support to the tissue. 4 Bioglass R (BG), a biodegradable bioactive silicate glass (46.1 mol % SiO 2 , 26.9 mol % CaO, 24.2 mol % Na 2 O, 2.6 mol % P 2 O 5 ), was the first material capable of bonding with bone by the formation of an apatite layer that forms on its surface after grafting. 5 A second mode of action is its dissolution products (in particular silica), which enhance osteoblast activities, whilst promoting extra- * Corresponding author, E-mail: julian.r.jones@imperial.ac cellular matrix (ECM) production and cells differentiation. 6-8 3D scaffolds have been produced from bioactive glasses, with interconnected pore networks in the form of foams 9,10 or 3-D printed structures 11 , which can take load in compression, up to 150 MPa when 3D printed. 12 ...

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“…Vinculin expression did not show a significant modulation ( Figure 4 D). Given that various stimuli have been shown to induce reversible dissipation of vinculin from focal adhesions and internalization of vinculin into vesicles [ 21 ], we investigated, by immunofluorescence, the distribution of vinculin in VECs. The same ECM substrates that showed low adherence of VECs, namely, collagen I, fibronectin, or vitronectin, were used for plate coating before cultivating VECs and treating cells with high glucose or direct interaction with monocytes.…”

Section: Resultsmentioning

confidence: 99%

The Specific Molecular Changes Induced by Diabetic Conditions in Valvular Endothelial Cells and upon Their Interactions with Monocytes Contribute to Endothelial Dysfunction

Tucureanu,

Ciortan,

Macarie

et al. 2024

IJMS

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Aortic valve disease (AVD) represents a global public health challenge. Research indicates a higher prevalence of diabetes in AVD patients, accelerating disease advancement. Although the specific mechanisms linking diabetes to valve dysfunction remain unclear, alterations of valvular endothelial cells (VECs) homeostasis due to high glucose (HG) or their crosstalk with monocytes play pivotal roles. The aim of this study was to determine the molecular signatures of VECs in HG and upon their interaction with monocytes in normal (NG) or high glucose conditions and to propose novel mechanisms underlying valvular dysfunction in diabetes. VECs and THP-1 monocytes cultured in NG/HG conditions were used. The RNAseq analysis revealed transcriptomic changes in VECs, in processes related to cytoskeleton regulation, focal adhesions, cellular junctions, and cell adhesion. Key molecules were validated by qPCR, Western blot, and immunofluorescence assays. The alterations in cytoskeleton and intercellular junctions impacted VEC function, leading to changes in VECs adherence to extracellular matrix, endothelial permeability, monocyte adhesion, and transmigration. The findings uncover new molecular mechanisms of VEC dysfunction in HG conditions and upon their interaction with monocytes in NG/HG conditions and may help to understand mechanisms of valvular dysfunction in diabetes and to develop novel therapeutic strategies in AVD.

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Physiologically induced restructuring of focal adhesions causes mobilization of vinculin by a vesicular endocytic recycling pathway

Cited by 6 publications

References 33 publications

Roles of Endocytic Processes and Early Endosomes on Focal Adhesion Dynamics in MDA-MB-231 Cells

Roles of Endocytic Processes and Early Endosomes on Focal Adhesion Dynamics in MDA-MB-231 Cells

Ductile silica/methacrylate hybrids for bone regeneration

The Specific Molecular Changes Induced by Diabetic Conditions in Valvular Endothelial Cells and upon Their Interactions with Monocytes Contribute to Endothelial Dysfunction

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