Abstract:Focal adhesion proteins link cell surface integrins and intracellular actin stress fibers and therefore play an important role in mechanotransduction and cell motility. When endothelial cells are subjected to cyclic mechanical strain, time-lapse imaging revealed that cells underwent significant morphological changes with their resultant long axes aligned away from the strain direction. To explore how this response is regulated by focal adhesion-associated proteins the expression levels of paxillin, focal adhes… Show more
“…Vimentin-GFP-transfected cells fixed and stained for vinculin-rich FAs (Figure 3), reveal that elongated cells (Figure 3(a)) had fewer FAs than flat cells (Figure 3(b)). This results in an inability to transmit forces from the FAs into the CSK [25]. It has also been postulated that fibroblast cells of this morphology have lower substrate-adhesion (which is consistent with our study) [26].…”
Section: Resultssupporting
confidence: 91%
“…It was observed that elongated cells have far fewer FAs than flat adherent cells. Thus it thought to reduce cell attachment to the substrate [25,26], which, combined with the reduced Young's Modulus of the elongated cells, allows for increased deformation of the IF CSK.…”
The intermediate filament (IF) cytoskeleton plays an important role in integrating biomechanical pathways associated with the actin and microtubule cytoskeleton. Vimentin is a type III IF protein commonly found in fibroblast cells and plays a role in transmitting forces through the cytoskeleton. Employing simultaneous laser scanning confocal and atomic force microscopy (AFM), we developed a methodology to quantify the deformation of the GFP-vimentin-labeled IF cytoskeleton as a function of time in response to force application by the AFM. Over short times (seconds), IFs deformed rapidly and transmitted force throughout the entire cell in a highly complex and anisotropic fashion. After several minutes, mechanically induced displacements of IFs resemble basal movements. In well-adhered cells the deformation of IFs is highly anisotropic as they tend to deform away from the longitudinal axis of the cell. This study demonstrates that simultaneous AFM and LSCM can be employed to track the deformation and dissipation of force through the IF cytoskeleton.
“…Vimentin-GFP-transfected cells fixed and stained for vinculin-rich FAs (Figure 3), reveal that elongated cells (Figure 3(a)) had fewer FAs than flat cells (Figure 3(b)). This results in an inability to transmit forces from the FAs into the CSK [25]. It has also been postulated that fibroblast cells of this morphology have lower substrate-adhesion (which is consistent with our study) [26].…”
Section: Resultssupporting
confidence: 91%
“…It was observed that elongated cells have far fewer FAs than flat adherent cells. Thus it thought to reduce cell attachment to the substrate [25,26], which, combined with the reduced Young's Modulus of the elongated cells, allows for increased deformation of the IF CSK.…”
The intermediate filament (IF) cytoskeleton plays an important role in integrating biomechanical pathways associated with the actin and microtubule cytoskeleton. Vimentin is a type III IF protein commonly found in fibroblast cells and plays a role in transmitting forces through the cytoskeleton. Employing simultaneous laser scanning confocal and atomic force microscopy (AFM), we developed a methodology to quantify the deformation of the GFP-vimentin-labeled IF cytoskeleton as a function of time in response to force application by the AFM. Over short times (seconds), IFs deformed rapidly and transmitted force throughout the entire cell in a highly complex and anisotropic fashion. After several minutes, mechanically induced displacements of IFs resemble basal movements. In well-adhered cells the deformation of IFs is highly anisotropic as they tend to deform away from the longitudinal axis of the cell. This study demonstrates that simultaneous AFM and LSCM can be employed to track the deformation and dissipation of force through the IF cytoskeleton.
“…Because previous studies have shown that cyclic tensile forces can affect orientation at the endothelial cellular level (27,28), we now set out to also examine their effect on vessel network morphology and orientation. To this end, 3D Gelfoam scaffolds cocultured with ECs and fibroblasts for 4 d were exposed to uniaxial cyclic stretching (10% strain and 1-Hz frequency).…”
Understanding the forces controlling vascular network properties and morphology can enhance in vitro tissue vascularization and graft integration prospects. This work assessed the effect of uniaxial cell-induced and externally applied tensile forces on the morphology of vascular networks formed within fibroblast and endothelial cell-embedded 3D polymeric constructs. Force intensity correlated with network quality, as verified by inhibition of force and of angiogenesis-related regulators. Tensile forces during vessel formation resulted in parallel vessel orientation under static stretching and diagonal orientation under cyclic stretching, supported by angiogenic factors secreted in response to each stretch protocol. Implantation of scaffolds bearing network orientations matching those of host abdominal muscle tissue improved graft integration and the mechanical properties of the implantation site, a critical factor in repair of defects in this area. This study demonstrates the regulatory role of forces in angiogenesis and their capacities in vessel structure manipulation, which can be exploited to improve scaffolds for tissue repair.
“…In the previous studies, the carcinogenesis of human CRC is related to the FAK destabilization [3,10,11] . By treatment of lactate-bound Ca 2+ , the full length of FAK is cleaved into a 90 kDa N-FAK protein (FERM domain).…”
Carcinogenic induction in a colon occurs through a sequence of events leading to metastasis that involved various oncogenic proteins. Focal adhesion kinase (FAK) regulates metastatic adhesion of carcinoma cells, and it has recognized as a potential therapeutic target toThe intracellular calcium (iCa 2+ ) ion is an important signaling molecule that modulates numerous cellular processes in cancer [1] . Ca 2+ permeable channels such as stored-operated calcium channels, transient receptor potential channels and the calcium release activated channel protein 1 are involved in the iCa 2+ homeostasis. Remodeling or deregulation of iCa 2+ homeostasis in cancer cells causes changes in cancer progression [2] . Focal adhesion kinase (FAK) plays a critical role in colon cancer progression, and it involved in cancer cell motility [3] . Calpain is a family of intracellular cysteine protease whose potentially induces cleavage of focal adhesion proteins depend on an increase in the iCa 2+ level. Calpain-mediated FAK degradation is critical in cancer cell motility [4] . The scaffolding function of FAK brings calpain into a focal adhesion complex, resulting in phosphorylation of calpain [5] . This phosphorylation leads to the activation of calpain, and then it leading to adhesion complex turnover and cell migration. Therefore, metastatic tumors contain higher level of calpain than non-metastatic tumors comparatively [6] . However, the impact of Ca 2+ supplementation has not been clearly understood on cancer metastasis. Based upon the results on direct exposure of lactate-bound Ca 2+ in colorectal cancer (CRC) cells, we highlight the increased motility of CRC cells that focused on the calpain-FAK pathway to understand underlying molecular mechanisms of human CRC cell motility by iCa 2+ [7] . It is well-known that lactate is highly utilized in the oxygenated cancer cells. Utilization of the lactate-bound Ca 2+ originated from this cancer's characteristic capacity to induce an increase in the Ca 2+ influx via lactate channel [8] .
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.