Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Dao, Lu; Gonnermann, Carina; Franz, Clemens M.

doi:10.1002/jmr.2303

Cited by 23 publications

(22 citation statements)

References 41 publications

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“…In mammalian cells, single receptor-ligand interactions - such as those between surface membrane proteins and extracellular matrix components - can generate forces up to 200 pN [34] and cooperativity within focal adhesions can increase the adhesion forces several hundred fold [24], [35], [36]. We found single steps during the detachment of cells with mean values of about 100 pN for wild type 0 hr cells (Fig.…”

Section: Discussionmentioning

confidence: 68%

Cell Substratum Adhesion during Early Development of Dictyostelium discoideum

et al. 2014

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Vegetative and developed amoebae of Dictyostelium discoideum gain traction and move rapidly on a wide range of substrata without forming focal adhesions. We used two independent assays to quantify cell-substrate adhesion in mutants and in wild-type cells as a function of development. Using a microfluidic device that generates a range of hydrodynamic shear stress, we found that substratum adhesion decreases at least 10 fold during the first 6 hr of development of wild type cells. This result was confirmed using a single-cell assay in which cells were attached to the cantilever of an atomic force probe and allowed to adhere to untreated glass surfaces before being retracted. Both of these assays showed that the decrease in substratum adhesion was dependent on the cAMP receptor CAR1 which triggers development. Vegetative cells missing talin as the result of a mutation in talA exhibited slightly reduced adhesive properties compared to vegetative wild-type cells. In sharp contrast to wild-type cells, however, these talA mutant cells did not show further reduction of adhesion during development such that after 5 hr of development they were significantly more adhesive than developed wild type cells. In addition, both assays showed that substrate adhesion was reduced in 0 hr cells when the actin cytoskeleton was disrupted by latrunculin. Consistent with previous observations, substrate adhesion was also reduced in 0 hr cells lacking the membrane proteins SadA or SibA as the result of mutations in sadA or sibA. However, there was no difference in the adhesion properties between wild type AX3 cells and these mutant cells after 6 hr of development, suggesting that neither SibA nor SadA play an essential role in substratum adhesion during aggregation. Our results provide a quantitative framework for further studies of cell substratum adhesion in Dictyostelium.

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Section: Discussionmentioning

confidence: 68%

Cell Substratum Adhesion during Early Development of Dictyostelium discoideum

et al. 2014

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“…S4A). This demonstrates that when delay times are used between sequential adhesion cycles, the cell adhesion shows no memory of previous adhesion events (Zarnitsyna et al, 2007;Friedrichs et al, 2010;Dao et al, 2013). Therefore, repetitive cell adhesion measurements were treated as uncorrelated events.…”

Section: Adhesion Time and Cytoskeletal Linking Modulate Alcam-mediatmentioning

confidence: 99%

Dynamic coupling of ALCAM to the actin cortex strengthens cell adhesion to CD6

Riet

Helenius

Strohmeyer

et al. 2014

Journal of Cell Science

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At the immunological synapse, the activated leukocyte cell adhesion molecule (ALCAM) on a dendritic cell (DC) and CD6 molecules on a T cell contribute to sustained DC-T-cell contacts. However, little is known about how ALCAM-CD6 bonds resist and adapt to mechanical stress. Here, we combine single-cell force spectroscopy (SCFS) with total-internal reflection fluorescence microscopy to examine ALCAM-CD6-mediated cell adhesion. The combination of cells expressing ALCAM constructs with certain cytoplasmic tail mutations and improved SCFS analysis processes reveal that the affinity of ALCAM-CD6 bonds is not influenced by the linking of the intracellular domains of ALCAM to the actin cortex. By contrast, the recruitment of ALCAM to adhesion sites and the propensity of ALCAM to anchor plasma membrane tethers depend on actin cytoskeletal interactions. Furthermore, linking ALCAM to the actin cortex through adaptor proteins stiffens the cortex and strengthens cell adhesion. We propose a framework for how ALCAMs contribute to DC-T-cell adhesion, stabilize DC-T-cell contacts and form a mechanical link between CD6 and the actin cortex to strengthen cell adhesion at the immunological synapse.

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“…Pulling experiments have been employed to study the mechanical stability of single protein molecules or specific bonds of biological interest (Williams et al ., ; Bellani et al ., ). More recently, also, cell adhesion on functional substrates or new materials has been characterized by AFM experiments (Dao et al ., ; Canale et al ., ). A direct comparison between local properties and sample topography can be achieved operating in the so‐called Force Volume (FV) modality.…”

Section: Introductionmentioning

confidence: 98%

High resolution nanomechanical characterization of multi‐domain model membranes by fast Force Volume

Seghezza

Dante

Diaspro

et al. 2015

J of Molecular Recognition

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Plasma membrane is a complex structure, mainly composed by lipids and proteins, which plays a pivotal role in cell metabolism by regulating its selective permeability to ions and molecules. According to the "raft hypothesis", lipids in the bilayer are not forming a structurally passive solvent, but are rather organized in specific domains, which present different structural and functional characteristics. The mechanical properties of the lipid part of plasma membrane have been recently characterized through Atomic Force Microscopy, by analyzing the features of force vs distance curves collected on supported lipid bilayers (SLBs). In case of lipid domains sizing from tens to hundreds of nanometers, which mimic in a good way the lateral organization of real membranes, a high lateral resolution and a large number of curves are often required for properly expressing the complexity of the system, with a consequent exponential growth of acquisition and processing time. In this paper we propose a method, based on a recently developed high speed Force Volume technique and on home-built data processing software, for the mechanical characterization of nanostructured SLBs. With our software we have been able to process data set composed by tens of thousands of curves, collected with a spatial resolution ranging from 8 to 40 nm/pixel. Multiparametric maps and distribution histograms produced by our analysis allowed identifying a specific behavior for each lipid phase in the investigated model membranes, even in presence of nanosized features.

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Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Cited by 23 publications

References 41 publications

Cell Substratum Adhesion during Early Development of Dictyostelium discoideum

Cell Substratum Adhesion during Early Development of Dictyostelium discoideum

Dynamic coupling of ALCAM to the actin cortex strengthens cell adhesion to CD6

High resolution nanomechanical characterization of multi‐domain model membranes by fast Force Volume

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