No abstract

The actin cytoskeleton is organized into diverse meshworks and bundles that support many aspects of cell physiology. Understanding the self-assembly of these actin-based structures is essential for developing predictive models of cytoskeletal organization. Here we show that the competing kinetics of bundle formation with the onset of dynamic arrest arising from filament entanglements and cross-linking determine the architecture of reconstituted actin networks formed with α-actinin cross-links. Cross-link mediated bundle formation only occurs in dilute solutions of highly mobile actin filaments. As actin polymerization proceeds, filament mobility and bundle formation are arrested concomitantly. By controlling the onset of dynamic arrest, perturbations to actin assembly kinetics dramatically alter the architecture of biochemically identical samples. Thus, the morphology of reconstituted F-actin networks is a kinetically determined structure similar to those formed by physical gels and glasses. These results establish mechanisms controlling the structure and mechanics in diverse semi-flexible biopolymer networks.

We drive optically trapped microspheres through entangled F-actin at constant speeds and distances well beyond the linear regime, and measure the microscale force response of the entangled filaments during and following strain. Our results reveal a unique crossover to appreciable nonlinearity at a strain rate of [small gamma, Greek, dot above]c ≈ 3 s(-1) which corresponds remarkably well with the theoretical rate of relaxation of entanglement length deformations 1/τent. Above [small gamma, Greek, dot above]c, we observe stress stiffening which occurs over very short time scales comparable to the predicted timescale over which mesh size deformations relax. Stress softening then takes over, yielding to an effectively viscous regime over a timescale comparable to the entanglement length relaxation time, τent. The viscous regime displays shear thinning but with a less pronounced viscosity scaling with strain rate compared to flexible polymers. The relaxation of induced force on filaments following strain shows that the relative relaxation proceeds more quickly for increasing strain rates; and for rates greater than [small gamma, Greek, dot above]c, the relaxation displays a complex power-law dependence on time. Our collective results reveal that molecular-level nonlinear viscoelasticity is driven by non-classical dynamics of individual entanglement segments that are unique to semiflexible polymers.

We optically drive a microsphere at constant speed through entangled actin networks of 0.2 -1.4 mg/ml at rates faster than the critical rate controlling the onset of a nonlinear response. By measuring the resistive force exerted on the microsphere during and following strain we reveal a critical concentration c * 0.4 mg/ml for nonlinear features to emerge. For c > c * , entangled actin stiffens at short times with the degree of stiffening S and corresponding timescale t sti f f scaling with the entanglement tube density, i.e. S ∼ t sti f f ∼ d

We track the deformation of discrete entangled actin segments while simultaneously measuring the resistive force the deformed filaments exert in response to an optically driven microsphere. We precisely map the network deformation field to show that local microscale stresses can induce filament deformations that propagate beyond mesoscopic length scales (60 μm, >3 persistence lengths l p). We show that the filament persistence length controls the critical length scale at which distinct entanglement deformations become driven by collective network mechanics. Mesoscale propagation beyond l p is coupled with nonlinear local stresses arising from steric entanglements mimicking cross-links.