Shear Flow-Induced Detachment Kinetics of Dictyostelium discoideum Cells from Solid Substrate

Décavé, Emmanuel; Garrivier, Daniel; Bréchet, Y.; Fourcade, B.; Brückert, Franz

doi:10.1016/s0006-3495(02)75583-5

Cited by 88 publications

(94 citation statements)

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“…The direct effect of calcium on passive cell adhesion to glass was also investigated, measuring σ 50% as a function of Ca 2+ concentration in a radial flow chamber. This parameter is the shear stress at which half of the cells detach and is directly related to the interaction energy between adhesion protein complexes and the substrate (Décavé et al, 2002). Radial flow detachment assays were conducted in the presence of CIPC, to uncouple passive adhesion from cytoskeleton dynamics (Décavé et al, 2002;Décavé et al, 2003).…”

Section: Resultsmentioning

confidence: 99%

“…Vegetative cells were harvested during exponential growth phase at a density of 2-4ϫ10 6 cells ml -1 , pelleted by centrifugation (1000 g, 4°C, 4 minutes), washed twice in Sörensen phosphate buffer (SB: 2 mM Na 2 HPO 4 , 14.5 mM KH 2 PO 4 , pH 6.2) or MES-Na buffer (20 mM morpholino ethane sulfonic acid, adjusted to pH 6.2 with NaOH) and used immediately. Glass plates or coverslips were cleaned with a ionic detergent and etched for 5 minutes with 14.5 M NaOH as previously described (Décavé et al, 2002). New glass plates or coverslips were used for each experiment.…”

Section: Methodsmentioning

confidence: 99%

“…Cell adhesion to glass plates was measured in SB at different CaCl 2 concentrations in the presence of the cytoskeleton depolymerizing agent N-(3-chlorophenyl)isopropyl carbamate (CIPC) at a 2 μg ml -1 final concentration from a 10 mg ml -1 stock in DMSO, with a radial flow detachment assay (Décavé et al, 2002). Flow was applied during 15 minutes to complete the detachment kinetics.…”

Section: Cell Adhesion Measurementsmentioning

confidence: 99%

See 2 more Smart Citations

Calcium mobilization stimulatesDictyostelium discoideumshear-flow-induced cell motility

Fache

Dalous

Engelund

et al. 2005

Journal of Cell Science

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Application of hydrodynamic mild shear stress to adherent Dictyostelium discoideum vegetative cells triggers active actin cytoskeleton remodeling resulting in net cell movement along the flow. The average cell speed is strongly stimulated by external calcium (Ca2+, K50%=22 μM), but the directionality of the movement is almost unaffected. This calcium concentration is ten times higher than the one promoting cell adhesion to glass surfaces (K50%=2 μM). Addition of the calcium chelator EGTA or the Ca2+-channel blocker gadolinium (Gd3+) transiently stops cell movement. Monitoring the evolution of cell-surface contact area with time reveals that calcium stimulates cell speed by increasing the amplitude of both protrusion and retraction events at the cell edge, but not the frequency. As a consequence, with saturating external calcium concentrations, cells are sensitive to very low shear forces (20 pN; σ=0.1 Pa). Moreover, a null-mutant lacking the unique Gβ subunit does not respond to external Ca2+ changes (K50%>1000 μM), although the directionality of the movement is comparable with that of wild-type cells. Furthermore, cells lacking the inositol 1,4,5-trisphosphate receptor (IP3-receptor) exhibit a markedly reduced Ca2+ sensitivity. Thus, calcium release from internal stores and calcium entry through the plasma membrane modulate cell speed in response to shear stress.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Cell Adhesion Measurementsmentioning

confidence: 99%

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Calcium mobilization stimulatesDictyostelium discoideumshear-flow-induced cell motility

Fache

Dalous

Engelund

et al. 2005

Journal of Cell Science

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“…These mutants also lost shear-induced motility in the direction of fluid flow (Décavé et al, 2002;Décavé et al, 2003;Fache et al, 2005;Lombardi et al, 2008 , but not cAMP. This result is surprising for several reasons.…”

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confidence: 96%

Myosin heavy chain kinases play essential roles in Ca2+, but not cAMP, chemotaxis and the natural aggregation of Dictyostelium discoideum

Wessels

Lusche

Steimle

et al. 2012

Journal of Cell Science

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SummaryBehavioral analyses of the deletion mutants of the four known myosin II heavy chain (Mhc) kinases of Dictyostelium discoideum revealed that all play a minor role in the efficiency of basic cell motility, but none play a role in chemotaxis in a spatial gradient of cAMP generated in vitro. However, the two kinases MhckA and MhckC were essential for chemotaxis in a spatial gradient of Ca 2+ , shear-induced directed movement, and reorientation in the front of waves of cAMP during natural aggregation. The phenotypes of the mutants mhckA 2 and mhckC 2 were highly similar to that of the Ca 2+ channel/receptor mutant iplA 2 and the myosin II phosphorylation mutant 3XALA, which produces constitutively unphosphorylated myosin II. These results demonstrate that IplA, MhckA and MhckC play a selective role in chemotaxis in a spatial gradient of Ca 2+ , but not cAMP, and suggest that Ca 2+ chemotaxis plays a role in the orientation of cells in the front of cAMP waves during natural aggregation.

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“…Cells can overcome this limitation by relaying chemotactic signals, but chemical relay of directional information requires intricate orchestration and timing of signals (7,10,11). Shear flow is another approach to guiding cells unidirectionally over large distances, but shear flow is an active process that requires constant fluid flow at a controlled rate and viscosity (12,13). Surface nanotopography, such as ridges and grooves (14)(15)(16)(17) or aligned collagen fibers (18), can act as a primitive and ubiquitous guidance cue, but the symmetric structures used in prior studies only provided bidirectional guidance.…”

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confidence: 99%

Asymmetric nanotopography biases cytoskeletal dynamics and promotes unidirectional cell guidance

Sun

Driscoll

Guven

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Many biological and physiological processes depend upon directed migration of cells, which is typically mediated by chemical or physical gradients or by signal relay. Here we show that cells can be guided in a single preferred direction based solely on local asymmetries in nano/microtopography on subcellular scales. These asymmetries can be repeated, and thereby provide directional guidance, over arbitrarily large areas. The direction and strength of the guidance is sensitive to the details of the nano/microtopography, suggesting that this phenomenon plays a context-dependent role in vivo. We demonstrate that appropriate asymmetric nano/microtopography can unidirectionally bias internal actin polymerization waves and that cells move with the same preferred direction as these waves. This phenomenon is observed both for the pseudopoddominated migration of the amoeboid Dictyostelium discoideum and for the lamellipod-driven migration of human neutrophils. The conservation of this mechanism across cell types and the asymmetric shape of many natural scaffolds suggest that actin-wave-based guidance is important in biology and physiology.cell migration | actin waves | contact guidance | Dictyostelium discoideum | neutrophils D irected cell migration is essential for many critical biological and physiological processes (1), such as embryonic development (2), wound healing (3), immune response (4), and angiogenesis (5). Guidance of cells can be achieved through external gradients in properties such as chemical concentration (6, 7), substrate rigidity (8), and adhesion (9). The total distance over which gradients can guide cells is limited by the finite dynamic range of cellular sensing [i.e., guidance by a gradient between the front and back of each cell requires that the overall signal change significantly with the cell's position (Fig. 1A)]. Cells can overcome this limitation by relaying chemotactic signals, but chemical relay of directional information requires intricate orchestration and timing of signals (7, 10, 11). Shear flow is another approach to guiding cells unidirectionally over large distances, but shear flow is an active process that requires constant fluid flow at a controlled rate and viscosity (12, 13). Surface nanotopography, such as ridges and grooves (14-17) or aligned collagen fibers (18), can act as a primitive and ubiquitous guidance cue, but the symmetric structures used in prior studies only provided bidirectional guidance.Although previous work has implicated cytoskeletal structures [in particular, the alignment of stress fibers (16)] in similar contact guidance processes, we have recently shown that nanotopography also steers the dynamics of the cell's scaffolding by biasing actin polymerization waves (17). Intracellular dynamics involving the self-assembly of actin and actin's associated proteins into 3D, traveling waves that propel a cell forward through a sustained cycle of polymerization and depolymerization have recently been found to be ubiquitous in cell migration (19)(20)(21)(22). Given ...

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Shear Flow-Induced Detachment Kinetics of Dictyostelium discoideum Cells from Solid Substrate

Cited by 88 publications

References 40 publications

Calcium mobilization stimulatesDictyostelium discoideumshear-flow-induced cell motility

Calcium mobilization stimulatesDictyostelium discoideumshear-flow-induced cell motility

Myosin heavy chain kinases play essential roles in Ca2+, but not cAMP, chemotaxis and the natural aggregation of Dictyostelium discoideum

Asymmetric nanotopography biases cytoskeletal dynamics and promotes unidirectional cell guidance

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