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IntroductionPhotomodifiable azopolymer nanotopographies represent a powerful means of assessing how cells respond to rapid changes in the local microenvironment. However, previous studies have suggested that azopolymers are readily photomodified under typical fluorescence imaging conditions over much of the visible spectrum. Here we assess the stability of azopolymer nanoridges under 1-photon and 2-photon imaging over a broad range of wavelengths. MethodsAzopolymer nanoridges were created via microtransfer molding of master structures that were created using interference lithography. The effects of exposure to a broad range of wavelengths of light polarized parallel to the ridges were assessed on both a spinning-disk confocal microscope and a 2-photon fluorescence microscope. Experiments with liveDictyostelium discoideumcells were also performed using alternating cycles of 514-nm light for photomodification and 561-nm light for fluorescence imaging. Results and DiscussionWe find that for both 1-photon and 2-photon imaging, only a limited range of wavelengths of light leads to photomodification of the azopolymer nanotopography. These results indicate that nondestructive 1-photon and 2-photon fluorescence imaging can be performed over a considerably broader range of wavelengths than would be suggested by previous research. 

Abstract Plasticity and homeostatic mechanisms allow neural networks to maintain proper function while responding to physiological challenges. Despite previous work investigating morphological and synaptic effects of brain-derived neurotrophic factor (BDNF), the most prevalent growth factor in the central nervous system, how exposure to BDNF manifests at the network level remains unknown. Here we report that BDNF treatment affects rodent hippocampal network dynamics during development and recovery from glutamate-induced excitotoxicity in culture. Importantly, these effects are not obvious when traditional activity metrics are used, so we delve more deeply into network organization, functional analyses, and in silico simulations. We demonstrate that BDNF partially restores homeostasis by promoting recovery of weak and medium connections after injury. Imaging and computational analyses suggest these effects are caused by changes to inhibitory neurons and connections. From our in silico simulations, we find that BDNF remodels the network by indirectly strengthening weak excitatory synapses after injury. Ultimately, our findings may explain the difficulties encountered in preclinical and clinical trials with BDNF and also offer information for future trials to consider. 

Ena/VASP proteins are processive actin polymerases that are required throughout animal phylogeny for many morphogenetic processes, including axon growth and guidance. Here we use in vivo live imaging of morphology and actin distribution to determine the role of Ena in promoting the growth of the TSM1 axon of the Drosophila wing. Altering Ena activity causes stalling and misrouting of TSM1. Our data show that Ena has a substantial impact on filopodial morphology in this growth cone but exerts only modest effects on actin distribution. This is in contrast to the main regulator of Ena, Abl tyrosine kinase, which was shown previously to have profound effects on actin and only mild effects on TSM1 growth cone morphology. We interpret these data as suggesting that the primary role of Ena in this axon may be to link actin to morphogenetic processes of the plasma membrane, rather than for regulating actin organization itself. These data also suggest that a key role of Ena, acting downstream of Abl, may be to maintain consistent organization and reliable evolution of growth cone structure, even as Abl activity varies in response to guidance cues in the environment. 

Abstract Asymmetric nanotopography with sub-cellular dimensions has recently demonstrated the ability to provide a unidirectional bias in cell migration. The details of this guidance depend on the type of cell studied and the design of the nanotopography. This behavior is not yet well understood, so there is a need for a predictive description of cell migration on such nanotopography that captures both the initiation of migration, and the way cell migration evolves. Here, we employ a three-dimensional, physics-based model to study cell guidance on asymmetric nanosawteeth. In agreement with experimental data, our model predicts that asymmetric sawteeth lead to spontaneous motion. Our model demonstrates that the nanosawteeth induce a unidirectional bias in guidance direction that is dependent upon actin polymerization rate and sawtooth dimensions. Motivated by this model, an analysis of previously reported experimental data indicates that the degree of guidance by asymmetric nanosawteeth increases with the cell velocity. 

We numerically study the effect of inter-particle friction coefficient on the response to cyclical pure shear of spherical particles in three dimensions. We focus on the rotations and translations of grains and look at the spatial distribution of these displacements as well as their probability distribution functions. We find that with increasing friction, the shear band becomes thinner and more pronounced. At low friction, the amplitude of particle rotations is homogeneously distributed in the system and is therefore mostly independent from both the affine and non-affine particle translations. In contrast, at high friction, the rotations are strongly localized in the shear zone. This work shows the importance of studying the effects of inter-particle friction on the response of granular materials to cyclic forcing, both for a better understanding of how rotations correlate to translations in sheared granular systems, and due to the relevance of cyclic forcing for most real-world applications in planetary science and industry. 

Abstract Nanotopographic surfaces are a powerful tool for studying and controlling cell behavior. However, the fabrication of nanotopographic master patterns using conventional photolithography is expensive, which limits the range of designs that can be explored. In this study, a method is demonstrated for the photoreshaping of large‐area patterns of nanoridges. The original master pattern is created using conventional lithography, and an azopolymer replica is prepared using soft lithography. The manipulation of the nanoridges is achieved by projecting light with specific polarizations and exposure times, resulting in controllable widening, buckling, or removal of the ridges. The reprogrammed azopolymer master patterns can then be replicated, creating reproducible new nanotopographies that can be transferred into other materials using a molding procedure. Diffraction can be used for in situ monitoring of the reprogramming during exposure. Image‐analysis methods are used to characterize buckled ridges as a function of exposure time. The response of MCF10A epithelial cells are investigated to buckled nanoridges. A substantial impact of buckling on the dynamics and location of actin polymerization, as well as on the distribution and lengths of contiguous polymerized regions is also observed. 

The dynamic rearrangement of the actin cytoskeleton is an essential component of many mechanotransduction and cellular force generation pathways. Here we use periodic surface topographies with feature sizes comparable to those of in vivo collagen fibers to measure and compare actin dynamics for two representative cell types that have markedly different migratory modes and physiological purposes: slowly migrating epithelial MCF10A cells and polarizing, fast-migrating, neutrophil-like HL60 cells. Both cell types exhibit reproducible guidance of actin waves (esotaxis) on these topographies, enabling quantitative comparisons of actin dynamics. We adapt a computer-vision algorithm, optical flow, to measure the directions of actin waves at the submicron scale. Clustering the optical flow into regions that move in similar directions enables micron-scale measurements of actin-wave speed and direction. Although the speed and morphology of actin waves differ between MCF10A and HL60 cells, the underlying actin guidance by nanotopography is similar in both cell types at the micron and submicron scales.