Eukaryotic Cell Migration Research Articles

Modeling Cell Movement and Chemotaxis Using Pseudopod-Based Feedback

A computational framework is presented for the simulation of eukaryotic cell migration and chemotaxis. An empirical pattern formation model, based on a system of nonlinear reaction-diffusion equations, is approximated on an evolving cell boundary using an arbitrary Lagrangian Eulerian surface finite element method (ALE-SFEM). The solution state is used to drive a mechanical model of the protrusive and retractive forces exerted on the cell boundary. Movement of the cell is achieved using a level set method. Results are presented for cell migration with and without chemotaxis. The simulated behavior is compared with experimental results of migrating Dictyostelium discoideum cells.

Force Generation of Curved Actin Gels Characterized by Combined AFM-Epifluorescence Measurements

Polymerization of actin into branched filaments is the driving force behind active migration of eukaryotic cells and motility of intracellular organelles. The site-directed assembly of a polarized branched array forms an expanding gel that generates the force that pushes the membrane. Here, we use atomic force microscopy to understand the relation between actin polymerization and the produced force. Functionalized spherical colloidal probes of varying size and curvature are attached to the atomic force microscopy cantilever and initiate the formation of a polarized actin gel in a solution mimicking the in vivo context. The gel growth is recorded by epifluorescence microscopy both against the cantilever and in the perpendicular (lateral) nonconstrained direction. In this configuration, the gel growth stops simultaneously in both directions at the stall force, which corresponds to a pressure of 0.15 nN/μm2. The results show that the growth of the gel is limited laterally, in the absence of external force, by internal mechanical stresses resulting from a combination of the curved geometry and the molecular mechanism of site-directed assembly of a cohesive branched filament array.

Spatial Coordination of Actin Polymerization and ILK–Akt2 Activity during Endothelial Cell Migration

Eukaryotic cell migration proceeds by cycles of protrusion, adhesion, and contraction, regulated by actin polymerization, focal adhesion assembly, and matrix degradation. However, mechanisms coordinating these processes remain largely unknown. Here, we show that local regulation of thymosin-beta4 (Tbeta4) binding to actin monomer (G-actin) coordinates actin polymerization with metalloproteinase synthesis to promote endothelial cell motility. In particular and quite unexpectedly, FRET analysis reveals diminished interaction between Tbeta4 and G-actin at the cell leading edge despite their colocalization there. Profilin-dependent dissociation of G-actin-Tbeta4 complexes simultaneously liberates actin for filament assembly and facilitates Tbeta4 binding to integrin-linked kinase (ILK) in the lamellipodia. Tbeta4-ILK complexes then recruit and activate Akt2, resulting in matrix metalloproteinase-2 production. Thus, the actin-Tbeta4 complex constitutes a latent coordinating center for cell migratory behavior, allowing profilin to initiate a cascade of events at the leading edge that couples actin polymerization to matrix degradation.

Quantitative elucidation of a distinct spatial gradient-sensing mechanism in fibroblasts

Migration of eukaryotic cells toward a chemoattractant often relies on their ability to distinguish receptor-mediated signaling at different subcellular locations, a phenomenon known as spatial sensing. A prominent example that is seen during wound healing is fibroblast migration in platelet-derived growth factor (PDGF) gradients. As in the well-characterized chemotactic cells Dictyostelium discoideum and neutrophils, signaling to the cytoskeleton via the phosphoinositide 3-kinase pathway in fibroblasts is spatially polarized by a PDGF gradient; however, the sensitivity of this process and how it is regulated are unknown. Through a quantitative analysis of mathematical models and live cell total internal reflection fluorescence microscopy experiments, we demonstrate that PDGF detection is governed by mechanisms that are fundamentally different from those in D. discoideum and neutrophils. Robust PDGF sensing requires steeper gradients and a much narrower range of absolute chemoattractant concentration, which is consistent with a simpler system lacking the feedback loops that yield signal amplification and adaptation in amoeboid cells.

Quantifying Lamella Dynamics of Cultured Cells by SACED, a New Computer-Assisted Motion Analysis

Formation of lamellipodia and the retraction of ruffles are essential activities during motility and migration of eukaryotic cells. We have developed a computer-assisted stroboscopic method for the continuous observation of cell dynamics (stroboscopic analysis of cell dynamics, SACED) that allows one to analyze changes in lamellipodia protrusion and ruffle retraction with high resolution in space and time. To demonstrate the potential of this method we analyzed keratinocytes in culture, unstimulated or stimulated with epidermal growth factor (EGF), which is known to induce cell motility and migration. Keratinocytes stimulated with EGF exhibited a 2.6-fold increase in their migration velocity, which coincided with enhanced ruffle retraction velocity (144%) and increased ruffle frequency (135%) compared to control cells. We also recorded an enhanced frequency of lamellipodia (135%), whereas the velocity of lamellipodia protrusion remained unchanged. These results on ruffle and lamellipodia dynamics in epidermal cells show that SACED is at least equal to established methods in terms of accuracy. SACED is, however, advantageous concerning resolution in time and therefore allows one to analyze the activity of lamellipodia and ruffles in as yet unknown detail. Moreover, SACED offers two opportunities that render this technique superior to established methods: First, several parameters relevant to cell motility can be analyzed simultaneously. Second, a large number of cells can conveniently be examined, which facilitates the compilation of statistically significant data.

The directed migration of eukaryotic cells.

The in vivo migration of eukaryotic cells over surfaces is important to a wide range of physiological processes, such as the development of the embryo, defense against infections, wound healing, the homing of lym­ phocytes to lymphoid organs, and tumor metastases (cf Trinkaus 1 984). Despite much study, the directed migration of cells is not yet well under­ stood, no doubt because it is indeed a complex problem. Part of the complexity is due to the great variety of locomotory phenotypes that are expressed by different types of motile cells. Many different cells can migrate under appropriate circumstances: In this article we refer to the directed migration of fibroblasts, endothelial cells, epithelial cells, macrophages, granulocytes, fish epidermal keratocytes, and amebae. Many of these cell types show distinct locomotory characteristics. However, most inves­ tigators agree that common mechanisms must operate in all cell migration.

The Directed Migration of Eukaryotic Cells

The Directed Migration of Eukaryotic Cells

Membrane insertion at the leading edge of motile fibroblasts.

We are concerned with the mechanisms involved in the directed migration of eukaryotic cells. Previously we found that, inside cells at the edge of an experimental wound, the Golgi apparatus and the microtubule-organizing center were rapidly repositioned forward of the nucleus in the direction of subsequent cell migration into the wound. This repositioning was proposed to serve the purpose of introducing new membrane mass at the leading edge of the cell, by directing Golgi apparatus-derived vesicles bound for the plasma membrane to that edge. We now provide evidence to support this proposal. Cultured fibroblastic cells at the edge of a wound were infected with a temperature-sensitive mutant (0-45) of vesicular stomatitis virus. It is known that the G-protein, an integral membrane protein of the virus, is synthesized and remains in the rough endoplasmic reticulum at the nonpermissive temperature, but when the infected cells are shifted to the permissive temperature, the G-protein moves through the Golgi apparatus to the plasma membrane. By immunofluorescence microscopy, we here show that the first appearance of the G-protein at the cell surface corresponds to the leading edge of the motile cell. These observations are incorporated into a coherent scheme for the mechanisms involved in cell migration.