Biomedical subjects

Hans-Günther Döbereiner

Publications and source records attributed to Hans-Günther Döbereiner.

9 recordsLinked to original sources

PubMed2006

Lateral membrane waves constitute a universal dynamic pattern of motile cells.

We have monitored active movements of the cell circumference on specifically coated substrates for a variety of cells including mouse embryonic fibroblasts and T cells, as well as wing disk cells from fruit flies. Despite having different functions and being from multiple phyla, these cell types share a common spatiotemporal pattern in their normal membrane velocity; we show that protrusion and retraction events are organized in lateral waves along the cell membrane. These wave patterns indicate both spatial and temporal long-range periodic correlations of the actomyosin gel.

Actomyosin↗

PubMed2006

Continuous membrane-cytoskeleton adhesion requires continuous accommodation to lipid and cytoskeleton dynamics.

The plasma membrane of most animal cells conforms to the cytoskeleton and only occasionally separates to form blebs. Previous studies indicated that many weak interactions between cytoskeleton and the lipid bilayer kept the surfaces together to counteract the normal outward pressure of cytoplasm. Either the loss of adhesion strength or the formation of gaps in the cytoskeleton enables the pressure to form blebs. Membrane-associated cytoskeleton proteins, such as spectrin and filamin, can control the movement and aggregation of membrane proteins and lipids, e.g., phosphoinositol phospholipids (PIPs), as well as blebbing. At the same time, lipids (particularly PIPs) and membrane proteins affect cytoskeleton and signaling dynamics. We consider here the roles of the major phosphatidylinositol-4,5-diphosphate (PIP2) binding protein, MARCKS, and PIP2 levels in controlling cytoskeleton dynamics. Further understanding of dynamics will provide important clues about how membrane-cytoskeleton adhesion rapidly adjusts to cytoskeleton and membrane dynamics.

Animals↗

PubMed2005

Force sensing and generation in cell phases: analyses of complex functions.

Cellular morphology is determined by motility, force sensing, and force generation that must be finely controlled in a dynamic fashion. Contractile and extensile functions are integrated with the overall cytoskeleton, including linkages from the cytoplasmic cytoskeleton to the extracellular matrix and other cells by force sensing. During development, as cells differentiate, variations in protein expression levels result in morphological changes. There are two major explanations for motile behavior: either cellular motility depends in a continuous fashion on cell composition or it exhibits phases wherein only a few protein modules are activated locally for a given time. Indeed, in support of the latter model, the quantification of cell spreading and other motile activities shows multiple distinct modes of behavior, which we term "phases" because there exist abrupt transitions between them. Cells in suspension have a basal level of motility that enables them to probe their immediate environment. After contacting a matrix-coated surface, they rapidly transition to an activated spreading phase. After the development of a significant contact area, the cells contract repeatedly to determine the rigidity of the substrate and then develop force on matrix contacts. When cells are fully spread, extension activity is significantly decreased and focal complexes start to assemble near the cell periphery. For each of these phases, there are significant differences in protein activities, which correspond to differences in function. Thus overall morphological change of a tissue is driven by chemical signals and force-dependent activation of one or more motile phases in limited cell regions for defined periods.

Animals↗

PubMed2004

Dynamic phase transitions in cell spreading.

We monitored isotropic spreading of mouse embryonic fibroblasts on fibronectin-coated substrates. Cell adhesion area versus time was measured via total internal reflection fluorescence microscopy. Spreading proceeds in well-defined phases. We found a power-law area growth with distinct exponents in three sequential phases, which we denote as basal, continuous, and contractile spreading. High resolution differential interference contrast microscopy was used to characterize local membrane dynamics at the spreading front. Fourier power spectra of membrane velocity reveal the sudden development of periodic membrane retractions at the transition from continuous to contractile spreading. We propose that the classification of cell spreading into phases with distinct functional characteristics and protein activity serves as a paradigm for a general program of a phase classification of cellular phenotype.

Animals↗

PubMed2004

Periodic lamellipodial contractions correlate with rearward actin waves.

Cellular lamellipodia bind to the matrix and probe its rigidity through forces generated by rearward F-actin transport. Cells respond to matrix rigidity by moving toward more rigid matrices using an unknown mechanism. In spreading and migrating cells we find local periodic contractions of lamellipodia that depend on matrix rigidity, fibronectin binding and myosin light chain kinase (MLCK). These contractions leave periodic rows of matrix bound beta3-integrin and paxillin while generating waves of rearward moving actin bound alpha-actinin and MLCK. The period between contractions corresponds to the time for F-actin to move across the lamellipodia. Shortening lamellipodial width by activating cofilin decreased this period proportionally. Increasing lamellipodial width by Rac signaling activation increased this period. We propose that an actin bound, contraction-activated signaling complex is transported locally from the tip to the base of the lamellipodium, activating the next contraction/extension cycle.

Actin Depolymerizing Factors↗

PubMed2004

Nanometer analysis of cell spreading on matrix-coated surfaces reveals two distinct cell states and STEPs.

When mouse embryonic fibroblasts in suspension contact a matrix-coated surface, they rapidly adhere and spread. Using total internal reflection fluorescence microscopy of dye-loaded fibroblasts to quantify cell-substrate contact, we found that increasing the surface matrix density resulted in faster spreading initiation whereas lamellipodial dynamics during spreading were unaltered. After spreading initiation, most cells spread in an anisotropic manner through stochastic, transient extension periods (STEPs) with approximately 30 STEPs over 10 min to reach an area of 1300 micro m(2) +/- 300 micro m(2). A second mode of spreading, increased in serum-deprived cells, lacked STEPs and spread in a rapid, isotropic manner for 1-4 min. This isotropic mode was characterized by a high rate of area increase, 340 micro m(2)/min with 78% of the cell edge extending. Anisotropic cells spread slower via STEPs, 126 micro m(2)/min with 34% of the edge extending. During the initial 2-4 min of fast, isotropic spreading, centripetal flow of actin was low (0.8 micro m/min) whereas in anisotropic cells it was high from early times (4.7 micro m/min). After initial isotropic spreading, rearward actin movement increased and isotropic cells displayed STEPs similar to anisotropic cells. Thus, the two cell states display dramatically different spreading whereas long-term motility is based on STEPs.

Actinin↗

PubMed2003

Advanced flicker spectroscopy of fluid membranes.

The bending elasticity of a fluid membrane is characterized by its modulus and spontaneous curvature. We present a new method, advanced flicker spectroscopy of giant nonspherical vesicles, which makes it possible to simultaneously measure both parameters for the first time. Our analysis is based on the generation of a large set of reference data from Monte Carlo simulations of randomly triangulated surfaces. As an example of the potential of the procedure, we monitor thermal trajectories of vesicle shapes and discuss the elastic response of zwitterionic membranes to transmembrane pH gradients. Our technique makes it possible to easily characterize membrane curvature as a function of environmental conditions.

Computer Simulation↗

PubMed2003

Diacylglycerol-rich domain formation in giant stearoyl-oleoyl phosphatidylcholine vesicles driven by phospholipase C activity.

We have studied the effect of phospholipase C from Bacillus cereus and Clostridium perfringens (alpha-toxin) on giant stearoyl-oleoyl phosphatidylcholine (SOPC) vesicles. Enzyme activity leads to a binary mixture of SOPC and the diacylglycerol SOG, which phase separates into a SOPC-rich bilayer phase and a SOG-rich isotropic bulk-like domain embedded within the membrane, as seen directly by phase contrast microscopy. After prolonged enzymatic attack, all bilayer membranes are transformed into an isotropic pure SOG phase as characterized by fluorescence microscopy, differential scanning calorimetry, fluorescence anisotropy measurements, and small angle x-ray scattering. These domains may have biological relevance, serving as storage compartments for hydrophobic molecules and/or catalyzing cellular signaling events at their boundaries. Furthermore, in the early stages of asymmetric enzymatic attack to the external monolayer of giant vesicles, we observe a transient coupling of the second-messenger diacylglycerol to membrane spontaneous curvature, which decreases due to enzyme activity, before domain formation and final vesicle collapse occurs.

Diglycerides↗

PubMed2002

Giant hexagonal superstructures in diblock-copolymer membranes.

We have observed polymersomes of high genus with their vesicle wall organized on the micrometer scale either in a double bilayer connected by a lattice of passages or a tubular network with hexagonal symmetry. Experimentally found shape classes are identified within a theoretical phase diagram based on the bending energy of the polymer membrane. Pronounced morphological changes could be induced and controlled by temperature.

Butadienes↗