637 research outputs found
Elastohydrodynamic study of actin filaments using fluorescence microscopy
We probed the bending of actin subject to external forcing and viscous drag.
Single actin filaments were moved perpendicular to their long axis in an
oscillatory way by means of an optically tweezed latex bead attached to one end
of the filaments. Shapes of these polymers were observed by epifluorescence
microscopy. They were found to be in agreement with predictions of semiflexible
polymer theory and slender-body hydrodynamics. A persistence length of m could be extracted.Comment: RevTex, 4 pages, 5 eps figs, submitted to PR
Spatial fluctuations at vertices of epithelial layers: quantification of regulation by Rho pathway
In living matter, shape fluctuations induced by acto-myosin are usually
studied in vitro via reconstituted gels, whose properties are controlled by
changing the concentrations of actin, myosin and cross-linkers. Such an
approach deliberately avoids to consider the complexity of biochemical
signaling inherent to living systems. Acto-myosin activity inside living cells
is mainly regulated by the Rho signaling pathway which is composed of multiple
layers of coupled activators and inhibitors. We investigate how such a pathway
controls the dynamics of confluent epithelial tissues by tracking the
displacements of the junction points between cells. Using a phenomenological
model to analyze the vertex fluctuations, we rationalize the effects of
different Rho signaling targets on the emergent tissue activity by quantifying
the effective diffusion coefficient, the persistence time and persistence
length of the fluctuations. Our results reveal an unanticipated correlation
between layers of activation/inhibition and spatial fluctuations within
tissues. Overall, this work connects the regulation via biochemical signaling
with mesoscopic spatial fluctuations, with potential application to the study
of structural rearrangements in epithelial tissues.Comment: 8 pages, 3 figure
Calculation of Forces at Focal Adhesions from Elastic Substrate Data: The Effect of Localized Force and the Need for Regularization
AbstractForces exerted by stationary cells have been investigated on the level of single focal adhesions by combining elastic substrates, fluorescence labeling of focal adhesions, and the assumption of localized force when solving the inverse problem of linear elasticity theory. Data simulation confirms that the inverse problem is ill-posed in the presence of noise and shows that in general a regularization scheme is needed to arrive at a reliable force estimate. Spatial and force resolution are restricted by the smoothing action of the elastic kernel, depend on the details of the force and displacement patterns, and are estimated by data simulation. Corrections arising from the spatial distribution of force and from finite substrate size are treated in the framework of a force multipolar expansion. Our method is computationally cheap and could be used to study mechanical activity of cells in real time
Trapping and Wiggling: Elastohydrodynamics of Driven Microfilaments
We present a general theoretical analysis of semiflexible filaments subject
to viscous drag or point forcing. These are the relevant forces in dynamic
experiments designed to measure biopolymer bending moduli. By analogy with the
``Stokes problems" in hydrodynamics (fluid motion induced by that of a wall
bounding a viscous fluid), we consider the motion of a polymer one end of which
is moved in an impulsive or oscillatory way. Analytical solutions for the
time-dependent shapes of such moving polymers are obtained within an analysis
applicable to small-amplitude deformations. In the case of oscillatory driving,
particular attention is paid to a characteristic length determined by the
frequency of oscillation, the polymer persistence length, and the viscous drag
coefficient. Experiments on actin filaments manipulated with optical traps
confirm the scaling law predicted by the analysis and provide a new technique
for measuring the elastic bending modulus. A re-analysis of several published
experiments on microtubules is also presented.Comment: RevTex, 24 pages, 15 eps figs, uses cite.sty, Biophysical
Synchronization of active mechanical oscillators by an inertial load
Motivated by the operation of myogenic (self-oscillatory) insect flight
muscle, we study a model consisting of a large number of identical oscillatory
contractile elements joined in a chain, whose end is attached to a damped
mass-spring oscillator. When the inertial load is small, the serial coupling
favors an antisynchronous state in which the extension of one oscillator is
compensated by the contraction of another, in order to preserve the total
length. However, a sufficiently massive load can sychronize the oscillators and
can even induce oscillation in situations where isolated elements would be
stable. The system has a complex phase diagram displaying quiescent,
synchronous and antisynchrononous phases, as well as an unsual asynchronous
phase in which the total length of the chain oscillates at a different
frequency from the individual active elements.Comment: 5 pages, 4 figures, To appear in Phys. Rev. Let
Focal Contacts as Mechanosensors: Externally Applied Local Mechanical Force Induces Growth of Focal Contacts by an Mdia1-Dependent and Rock-Independent Mechanism
The transition of cell–matrix adhesions from the initial punctate focal complexes into the mature elongated form, known as focal contacts, requires GTPase Rho activity. In particular, activation of myosin II–driven contractility by a Rho target known as Rho-associated kinase (ROCK) was shown to be essential for focal contact formation. To dissect the mechanism of Rho-dependent induction of focal contacts and to elucidate the role of cell contractility, we applied mechanical force to vinculin-containing dot-like adhesions at the cell edge using a micropipette. Local centripetal pulling led to local assembly and elongation of these structures and to their development into streak-like focal contacts, as revealed by the dynamics of green fluorescent protein–tagged vinculin or paxillin and interference reflection microscopy. Inhibition of Rho activity by C3 transferase suppressed this force-induced focal contact formation. However, constitutively active mutants of another Rho target, the formin homology protein mDia1 (Watanabe, N., T. Kato, A. Fujita, T. Ishizaki, and S. Narumiya. 1999. Nat. Cell Biol. 1:136–143), were sufficient to restore force-induced focal contact formation in C3 transferase-treated cells. Force-induced formation of the focal contacts still occurred in cells subjected to myosin II and ROCK inhibition. Thus, as long as mDia1 is active, external tension force bypasses the requirement for ROCK-mediated myosin II contractility in the induction of focal contacts. Our experiments show that integrin-containing focal complexes behave as individual mechanosensors exhibiting directional assembly in response to local force
The Force-Velocity Relation for Growing Biopolymers
The process of force generation by the growth of biopolymers is simulated via
a Langevin-dynamics approach. The interaction forces are taken to have simple
forms that favor the growth of straight fibers from solution. The
force-velocity relation is obtained from the simulations for two versions of
the monomer-monomer force field. It is found that the growth rate drops off
more rapidly with applied force than expected from the simplest theories based
on thermal motion of the obstacle. The discrepancies amount to a factor of
three or more when the applied force exceeds 2.5kT/a, where a is the step size
for the polymer growth. These results are explained on the basis of restricted
diffusion of monomers near the fiber tip. It is also found that the mobility of
the obstacle has little effect on the growth rate, over a broad range.Comment: Latex source, 9 postscript figures, uses psfig.st
Self-organized Beating and Swimming of Internally Driven Filaments
We study a simple two-dimensional model for motion of an elastic filament
subject to internally generated stresses and show that wave-like propagating
shapes which can propel the filament can be induced by a self-organized
mechanism via a dynamic instability. The resulting patterns of motion do not
depend on the microscopic mechanism of the instability but only of the filament
rigidity and hydrodynamic friction. Our results suggest that simplified
systems, consisting only of molecular motors and filaments could be able to
show beating motion and self-propulsion.Comment: 8 pages, 2 figures, REVTe
The Viscous Nonlinear Dynamics of Twist and Writhe
Exploiting the "natural" frame of space curves, we formulate an intrinsic
dynamics of twisted elastic filaments in viscous fluids. A pair of coupled
nonlinear equations describing the temporal evolution of the filament's complex
curvature and twist density embodies the dynamic interplay of twist and writhe.
These are used to illustrate a novel nonlinear phenomenon: ``geometric
untwisting" of open filaments, whereby twisting strains relax through a
transient writhing instability without performing axial rotation. This may
explain certain experimentally observed motions of fibers of the bacterium B.
subtilis [N.H. Mendelson, et al., J. Bacteriol. 177, 7060 (1995)].Comment: 9 pages, 4 figure
Entanglement, elasticity and viscous relaxation of actin solutions
We have investigated the viscosity and the plateau modulus of actin solutions
with a magnetically driven rotating disc rheometer. For entangled solutions we
observed a scaling of the plateau modulus versus concentration with a power of
7/5. The measured terminal relaxation time increases with a power 3/2 as a
function of polymer length. We interpret the entanglement transition and the
scaling of the plateau modulus in terms of the tube model for semiflexible
polymers.Comment: 5 pages, 4 figures, published versio
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