Filament Network Research Articles

A unique F-actin and junctional organization that maintains the corneal endothelial barrier

Introduction: The corneal endothelium is responsible for the correct hydration of the corneal stroma and for an adequate transport through this tissue. Corneal endothelial layer has a particular organization to maintain its curved shape and function. The corneal endothelium has low proliferative capacity and form a monolayer that is subjected to an even mechanical tension coming from the positive pressure of the aqueous humor. Preserving their barrier function under suboptimal conditions, such as corneal pathologies, age and transplantation, is essential for maintaining corneal transparency. Material and methods: We have investigated the structure and the proteins in charge of maintaining corneal endothelial barrier function by confocal microscopy and time lapse spinning disk microscopy in murine corneas ex vivo, and hepatic organoids. Results: We have characterized a novel filamentous actin network that organizes into radial structures arising from the center of the cell under the nucleus towards cell-cell junctions. This structure is also observed in spherical epithelial organoids whose cells are also exposed to positive luminal pressure. Due to the simplicity of the model, it can be easily implemented in any clinic which leads to increasing ADR and preventing CRC, but requires validation in large multicenter trials.

Oxidation and reduction of actin: Origin, impact in vitro and functional consequences in vivo

Actin is among the most abundant proteins in eukaryotic cells and assembles into dynamic filamentous networks regulated by many actin binding proteins. The actin cytoskeleton must be finely tuned, both in space and time, to fulfill key cellular functions such as cell division, cell shape changes, phagocytosis and cell migration. While actin oxidation by reactive oxygen species (ROS) at non-physiological levels are known for long to impact on actin polymerization and on the cellular actin cytoskeleton, growing evidence shows that direct and reversible oxidation/reduction of specific actin amino acids plays an important and physiological role in regulating the actin cytoskeleton. In this review, we describe which actin amino acid residues can be selectively oxidized and reduced in many different ways (e.g. disulfide bond formation, glutathionylation, carbonylation, nitration, nitrosylation and other oxidations), the cellular enzymes at the origin of these post-translational modifications, and the impact of actin redox modifications both in vitro and in vivo. We show that the regulated balance of oxidation and reduction of key actin amino acid residues contributes to the control of actin filament polymerization and disassembly at the subcellular scale and highlight how improper redox modifications of actin can lead to pathological conditions.

3D Morphology of Open Clusters in the Solar Neighborhood with Gaia EDR 3. II. Hierarchical Star Formation Revealed by Spatial and Kinematic Substructures

We identify members of 65 open clusters in the solar neighborhood using the machine-learning algorithm StarGO based on Gaia EDR3 data. After adding members of 20 clusters from previous studies we obtain 85 clusters, and study their morphology and kinematics. We classify the substructures outside the tidal radius into four categories: filamentary (f1) and fractal (f2) for clusters <100 Myr, and halo (h) and tidal tail (t) for clusters >100 Myr. The kinematical substructures of f1-type clusters are elongated; these resemble the disrupted cluster Group X. Kinematic tails are distinct in t-type clusters, especially Pleiades. We identify 29 hierarchical groups in four young regions (Alessi 20, IC 348, LP 2373, LP 2442); 10 among these are new. The hierarchical groups form filament networks. Two regions (Alessi 20, LP 2373) exhibit global orthogonal expansion (stellar motion perpendicular to the filament), which might cause complete dispersal. Infalling-like flows (stellar motion along the filament) are found in UBC 31 and related hierarchical groups in the IC 348 region. Stellar groups in the LP 2442 region (LP 2442 gp 1–5) are spatially well mixed but kinematically coherent. A merging process might be ongoing in the LP 2442 subgroups. For younger systems (≲30 Myr), the mean axis ratio, cluster mass, and half-mass–radius tend to increase with age values. These correlations between structural parameters may imply two dynamical processes occurring in the hierarchical formation scenario in young stellar groups: (1) filament dissolution and (2) subgroup mergers.

Structure of Arp2/3 complex at a branched actin filament junction resolved by single-particle cryo-electron microscopy

Arp2/3 complex nucleates branched actin filaments that provide pushing forces to drive cellular processes such as lamellipodial protrusion and endocytosis. Arp2/3 complex is intrinsically inactive, and multiple classes of nucleation promoting factors (NPFs) stimulate its nucleation activity. When activated by WASP family NPFs, the complex must bind to the side of a preexisting (mother) filament of actin to complete the nucleation process, ensuring that WASP-mediated activation creates branched rather than linear actin filaments. How actin filaments contribute to activation is currently not understood, largely due to the lack of high-resolution structures of activated Arp2/3 complex bound to the side of a filament. Here, we present the 3.9-Å cryo-electron microscopy structure of the Arp2/3 complex at a branch junction. The structure reveals contacts between Arp2/3 complex and the side of the mother actin filament that likely stimulate subunit flattening, a conformational change that allows the actin-related protein subunits in the complex (Arp2 and Arp3) to mimic filamentous actin subunits. In contrast, limited contact between the bottom half of the complex and the mother filament suggests that clamp twisting, a second major conformational change observed in the active state, is not stimulated by actin filaments, potentially explaining why actin filaments are required but insufficient to trigger nucleation during WASP-mediated activation. Along with biochemical and live-cell imaging data and molecular dynamics simulations, the structure reveals features critical for the interaction of Arp2/3 complex with actin filaments and regulated assembly of branched actin filament networks in cells.

Crowding-induced membrane remodeling: Interplay of membrane tension, polymer density, architecture

The plasma membrane hosts a wide range of biomolecules, mainly proteins and carbohydrates, that mediate cellular interactions with its environment. The crowding of such biomolecules regulates cellular morphologies and cellular trafficking. Recent discoveries have shown that the structure and density of cell surface polymers and hence the signaling machinery change with the state of the cell, especially in cancer progression. The alterations in membrane-attached glycocalyx and glycosylation of proteins and lipids are common features of cancer cells. The overexpression of glycocalyx polymers, such as mucin and hyaluronan, strongly correlates with cancer metastasis. Here, we present a mesoscale biophysics-based model that accounts for the shape regulation of membranes by crowding of membrane-attached biopolymer-glycocalyx and actin networks. Our computational model is based on the dynamically triangulated Monte Carlo model for membranes and coarse-grained representations of polymer chains. The model allows us to investigate the crowding-induced shape transformations in cell membranes in a tension- and graft polymer density-dependent manner. Our results show that the number of membrane protrusions and their shape depend on membrane tension, with higher membrane tension inducing more tubular protrusions than the vesicular shapes formed at low tension at high surface coverage of polymers. The shape transformations occur above the threshold density predicted by the polymer brush theory, but this threshold also depends on the membrane tension. Increasing the size of the polymer, either by changing the length or by adding side chains, is shown to increase the crowding-induced curvature. The effect of crowding is more prominent for flexible polymers than for semiflexible rigid polymers. We also present an extension of the model that incorporates properties of the actin-like filament networks and demonstrate how tubular structures can be generated by biopolymer crowding on the cytosolic side of cell membranes.

The eROSITA view of the Abell 3391/95 field: Case study from the Magneticum cosmological simulation

Context.Clusters of galaxies reside at the nodes of the cosmic web, interconnected by filamentary structures that contain tenuous diffuse gas, especially in the warm-hot phase. Galaxy clusters grow by mergers of smaller objects and gas that are mainly accreted through these large-scale filaments. For the first time, the large-scale cosmic structure and a long gas-emission filament have been captured by eROSITA on board the Spectrum-Roentgen-Gamma mission in a direct X-ray observation of the A3391/95 field.Aims.We investigate the assembly history of an A3391/95-like system of clusters and the thermo-chemical properties of the diffuse gas in it by connecting simulation predictions to the eROSITA observations with the aim to constrain the origin and nature of the gas in the pair-interconnecting bridge.Methods.We analysed the properties of a system resembling A3391/95, extracted from the (352h−1cMpc)3volume of the Magneticum Pathfinder cosmological simulations atz= 0.07. We tracked the main progenitors of the pair clusters and of surrounding groups back in time to study the assembly history of the system and its evolution.Results.Similarly to the observed A3391/95 system, the simulated cluster pair is embedded in a complex network of gas filaments, with structures aligned over more than 20 projected Mpc, and the whole region collapses towards the central overdense node. The spheres of influence (3 ×R200) of the two main clusters already overlap atz= 0.07, but their virial boundaries are still physically separated. The diffuse gas located in the interconnecting bridge closely reflects the warm-hot intergalactic medium, with a typical temperature of ~1 keV and an overdensityδ ~100 with respect to the mean baryon density of the Universe, and a lower enrichment level compared to the intra-cluster medium in clusters. We find that most of the bridge gas collapsed from directions roughly orthogonal to the intra-cluster gas accretion directions, and its origin is mostly unrelated to the two cluster progenitors. We find clear signatures in the surrounding groups of infall motion towards the pair, such as significant radial velocities and a slowdown of gas compared to dark matter. These findings further support the hypothesis that the Northern Clump (MCXC J0621.7-5242) cluster infalls along a cosmic gas filament towards Abell 3391 and might be merging with it.Conclusions.We conclude that in this configuration, the pair clusters of the A3391/95-like system are in a pre-merger phase and have not yet interacted. The diffuse gas in the interconnecting bridge is mostly warm filament gas and not tidally stripped cluster gas.

Preprint Highlight: Building the cytokinetic contractile ring in an early embryo: initiation as clusters of myosin II, anillin and septin, and visualization of a septin filament network.

Cytokinesis is driven by a contractile ring whose key components are well defined, yet how they are organized and how the contractile ring assembles and constricts remain unclear. This study combines confocal and superresolution imaging of endogenous contractile ring components in sea urchin embryos (in vivo) and isolated embryo cortices (ex vivo) to demonstrate that anillin and septin2 assemble into myosin II-positive clusters. The author surmises that these clusters are precursors to the contractile ring and observe septin filament nanostructure for the first time in an animal cell contractile ring. These insights into the organization of lesser studied yet conserved contractile ring proteins, anillin and septins, provide clues to their function and pave the way for similar studies in other animal species.

Desmoglein‐2 and Desmoplakin modulate Cell Spreading Dynamics via PDZ‐GEF2 and Rap1 Signaling

The desmosome is a cell‐cell adhesion complex which facilitates the mechanical stability of tissues and cell‐cell communication. Desmosome function depends upon a tripartite organizational structure wherein transmembrane cadherins (Desmoglein and Desmocollin) link adjacent cells in the extracellular space, armadillo proteins (Plakophilin and Plakoglobin) stabilize the intracellular plaque, and the cytolinker Desmoplakin (DP) connects the plaque to the intermediate filament network. In addition to their central role in maintaining cell‐cell junction integrity, desmosomal cadherins also coordinate biological processes such as proliferation, apoptosis, differentiation and cell migration. In our study, we sought to investigate the signaling mechanisms involved in control of actin architecture via desmosomal cadherins, using A431 cells lacking Desmoglein‐2 (Dsg2 KO), generated via CRISPR‐mediated knock‐out. Compared to control cells, Dsg2 KO cells displayed a significant increase in spreading area on both fibronectin and collagen. As these experiments were performed in singly spreading cells, these experiments have identified a novel cell‐autonomous, cell‐cell adhesion‐independent role for Dsg2 in regulation of cell spreading. Spreading changes in Dsg2 KO cells were dependent on Rap1 GTPase, as siRNA‐mediated knockdown of Rap1 or pharmacological inhibition of Rap1 in Dsg2 KO cells rescued the increase in spreading area. Rap1 activity was significantly increased in Dsg2KO cells, but no change was observed in the activity of other GTPases tested (Rho, Rac and Cdc42). Changes in Dsg2‐mediated cell spreading was also shown to require the Rap GEF PDZ‐GEF2 (but not PDZ‐GEF1 or other Rap GEFs), and PDZ‐GEF2 localization at the cell periphery markedly increases upon loss of Dsg2. Further investigation into signaling pathways known to affect cell spreading identified a role for TGF‐beta signaling, but neither Erk or p38 MAPK were involved. In singly‐spreading cells, the expression of other desmosomal proteins is unchanged in Dsg2 KO cells (mRNA and protein levels), and the localization of Plakophilin‐2, Plakoglobin and Desmoplakin is also not significantly perturbed (specifically in singly‐spreading cells). Nevertheless, siRNA‐mediated knockdown of DP was able to rescue Dg2KO‐dependent increases in cell spreading (while siRNA for Plakophilin‐2, Plakoglobin and Plakophilin‐3 did not). In addition, DP siRNA was also able to rescue changes in localization of PDZ‐GEF2 seen upon loss of Dsg2. Taken together, these data have identified novel cell‐cell adhesion independent role for Dsg2 and DP in mediating cell spreading via Rap1 signaling, which provides significant insight into the signaling mechanisms via which desmosomal cadherins control cell‐matrix attachment and cytoskeletal architecture.

Establishing neuronal polarity: microtubule regulation during neurite initiation.

The initiation of nascent projections, or neurites, from the neuronal cell body is the first stage in the formation of axons and dendrites, and thus a critical step in the establishment of neuronal architecture and nervous system development. Neurite formation relies on the polarized remodelling of microtubules, which dynamically direct and reinforce cell shape, and provide tracks for cargo transport and force generation. Within neurons, microtubule behaviour and structure are tightly controlled by an array of regulatory factors. Although microtubule regulation in the later stages of axon development is relatively well understood, how microtubules are regulated during neurite initiation is rarely examined. Here, we discuss how factors that direct microtubule growth, remodelling, stability and positioning influence neurite formation. In addition, we consider microtubule organization by the centrosome and modulation by the actin and intermediate filament networks to provide an up-to-date picture of this vital stage in neuronal development.

Regulation of Extracellular Matrix Gene Expression by Desmosomal Cadherins

Desmosomes are protein complexes crucial for maintaining cell‐cell adhesion and integrity of tissues. These complexes are made up of proteins from three families: transmembrane cadherins (Desmoglein and Desmocollin) link adjacent cells in the extracellular space, armadillo proteins (Plakophilin and Plakoglobin) stabilize the intracellular plaque, and the cytolinker Desmoplakin (DP) connects the plaque to the intermediate filament network. Desmosomal proteins have also been shown to coordinate gene expression pathways required for processes such as proliferation, differentiation and cell migration. In particular, several lines of evidence have linked desmosomal proteins to gene expression of extracellular matrix (ECM) proteins. Loss of Plakophilin‐2 or Desmoplakin causes increases in expression of fibronectin and collagen, while in contrast, loss of plakoglobin results in a significant decrease in expression of fibronectin. These data indicate that individual components of the desmosome control ECM gene expression via distinct cellular signaling networks. In our study, we sought to investigate the role of desmosomal cadherins in ECM gene expression, via use of A431 cells lacking Desmoglein‐2, generated via CRISPR‐mediated knock‐out. Compared to control cells, Dsg2 KO cells demonstrated a dramatic increase in expression of Fibronectin (Fn1) mRNA, and relatively minor changes in expression of Collagen 1 (Col1a1) and Collagen 2 (Col2a1) mRNA. An increase in expression of Fibronectin protein levels in Dsg2‐deficient cells was also observed via western blot. We show that loss of Dsg2 KO also caused a significant increase in mRNA expression of the pro‐fibrotic signaling molecule transforming growth factor beta 2 (Tgfb2), but not Tgfb1 or Tgfb3. Increased expression of Fn1, Col1a1, Col2a1 and Tgfb2 was also observed upon siRNA‐mediated knockdown of Dsg2 in both A431 cells and HaCaT keratinocytes, verifying that these changes are not clone‐specific or due to off‐target CRISPR effects. siRNA‐mediated knockdown of other desmosomal proteins could not rescue Fn1 or Tgfb2 increases in Dsg2 KO cells, indicating that changes are not due to mislocalization of other desmosomal components. An extensive analysis of signaling pathways known to be involved in regulation of ECM gene expression uncovered an important role for PI3‐Kinase and Akt signaling, as inhibition of either signaling protein resulted in a rescue of Dg2KO‐mediated increases in Fn1 gene expression. Moreover, we have documented a significant increase in phosphorylation of Akt in Dsg2‐deficient cells. Taken together, our study highlights a novel role for Dsg2 in mediating ECM gene expression via PI3‐K/Akt signaling, adding significant insight into the mechanisms by which desmosomal cadherins control the adhesive behavior of cancer cells.

A coarse-grained approach to model the dynamics of the actomyosin cortex

BackgroundThe dynamics of the actomyosin machinery is at the core of many important biological processes. Several relevant cellular responses such as the rhythmic compression of the cell cortex are governed, at a mesoscopic level, by the nonlinear interaction between actin monomers, actin crosslinkers, and myosin motors. Coarse-grained models are an optimal tool to study actomyosin systems, since they can include processes that occur at long time and space scales, while maintaining the most relevant features of the molecular interactions.ResultsHere, we present a coarse-grained model of a two-dimensional actomyosin cortex, adjacent to a three-dimensional cytoplasm. Our simplified model incorporates only well-characterized interactions between actin monomers, actin crosslinkers and myosin, and it is able to reproduce many of the most important aspects of actin filament and actomyosin network formation, such as dynamics of polymerization and depolymerization, treadmilling, network formation, and the autonomous oscillatory dynamics of actomyosin.ConclusionsWe believe that the present model can be used to study the in vivo response of actomyosin networks to changes in key parameters of the system, such as alterations in the attachment of actin filaments to the cell cortex.

Adenomatous Polyposis Coli (APC) in cell migration

Adenomatous Polyposis Coli (APC) protein is mostly known as a tumor suppressor that regulates Wnt signaling, but is also an important cytoskeletal protein. Mutations in the APC gene are linked to colorectal cancer and various neurological disorders and intellectual disabilities. Cytoskeletal functions of APC appear to have significant contributions to both types of these disorders. As a cytoskeletal protein, APC can regulate both actin and microtubule cytoskeletons, which together form the main machinery for cell migration. As APC is a multifunctional protein with numerous interaction partners, the complete picture of how APC regulates cell motility is still unavailable. However, some molecular mechanisms begin to emerge. Here, we review available information about roles of APC in cell migration and propose a model explaining how microtubules, using APC as an intermediate, can initiate leading edge protrusion in response to external signals by stimulating Arp2/3 complex-dependent nucleation of branched actin filament networks via a series of intermediate events.

Myosin II proteins are required for organization of calcium-induced actin networks upstream of mitochondrial division.

The formin INF2 polymerizes a calcium-activated cytoplasmic network of actin filaments, which we refer to as calcium-induced actin polymerization (CIA). CIA plays important roles in multiple cellular processes, including mitochondrial dynamics and vesicle transport. Here, we show that nonmuscle myosin II (NMII) is activated within 60 s of calcium stimulation and rapidly recruited to the CIA network. Knockout of any individual NMII in U2OS cells affects the organization of the CIA network, as well as three downstream effects: endoplasmic-reticulum-to-mitochondrial calcium transfer, mitochondrial Drp1 recruitment, and mitochondrial division. Interestingly, while NMIIC is the least abundant NMII in U2OS cells (>200-fold less than NMIIA and >10-fold less than NMIIB), its knockout is equally deleterious to CIA. On the basis of these results, we propose that myosin II filaments containing all three NMII heavy chains exert organizational and contractile roles in the CIA network. In addition, NMIIA knockout causes a significant decrease in myosin regulatory light chain levels, which might have additional effects.

Intermediate Filaments in Cellular Mechanoresponsiveness: Mediating Cytoskeletal Crosstalk From Membrane to Nucleus and Back.

The mammalian cytoskeleton forms a mechanical continuum that spans across the cell, connecting the cell surface to the nucleus via transmembrane protein complexes in the plasma and nuclear membranes. It transmits extracellular forces to the cell interior, providing mechanical cues that influence cellular decisions, but also actively generates intracellular forces, enabling the cell to probe and remodel its tissue microenvironment. Cells adapt their gene expression profile and morphology to external cues provided by the matrix and adjacent cells as well as to cell-intrinsic changes in cytoplasmic and nuclear volume. The cytoskeleton is a complex filamentous network of three interpenetrating structural proteins: actin, microtubules, and intermediate filaments. Traditionally the actin cytoskeleton is considered the main contributor to mechanosensitivity. This view is now shifting owing to the mounting evidence that the three cytoskeletal filaments have interdependent functions due to cytoskeletal crosstalk, with intermediate filaments taking a central role. In this Mini Review we discuss how cytoskeletal crosstalk confers mechanosensitivity to cells and tissues, with a particular focus on the role of intermediate filaments. We propose a view of the cytoskeleton as a composite structure, in which cytoskeletal crosstalk regulates the local stability and organization of all three filament families at the sub-cellular scale, cytoskeletal mechanics at the cellular scale, and cell adaptation to external cues at the tissue scale.

Shear stress regulates the SNAP23-mediated endothelial secretion of VWF through the GPR68/PKA/vimentin mechanotransduction pathway

Von Willebrand Factor (VWF) can promote platelet adhesion to the post-atherosclerotic regions to initiate thrombosis. The synthesis and secretion of VWF are important functions of endothelial cells (ECs). However, the mechanism through which blood flow regulates endothelial secretion of VWF remains unclear. We utilized a parallel-plate flow apparatus to apply fluid shear stress to human umbilical vein endothelial cells (HUVECs). Compared with pulsatile shear stress that mimics laminar flow in the straight parts of arteries or upstream of atherosclerotic stenosis sites, short-term exposure to oscillatory shear stress (OS) that mimics disturbed flow increased VWF secretion independent of affecting synaptosomal-associated protein 23 (SNAP23) expression and promoted the translocation of SNAP23 to the cell membrane. Vimentin associated with SNAP23, and this association was enhanced by OS or histamine. Acrylamide, a reagent that disrupts vimentin intermediate filaments, prevented histamine/OS-induced SNAP23 translocation, as well as VWF secretion. Immunofluorescence analysis revealed that the polarity of the vimentin intermediate filament network decreased after stimulation with histamine or OS. In addition, inhibition of protein kinase A (PKA) or G protein coupled receptor 68 (GPR68) eliminated the histamine/OS-induced phosphorylation of vimentin at Ser38 and secretion of VWF. Furthermore, syntaxin 7 might assist with the translocation of SNAP23 to the cell membrane, thus playing a role in promoting VWF secretion. The GPR68/PKA/vimentin signaling pathway may represent a novel mechanism for the regulation of SNAP23-mediated VWF secretion by ECs under OS and provide strategies for the prevention of atherosclerosis-related thrombosis.

Parallel kinase pathways stimulate actin polymerization at depolarized mitochondria

Mitochondrial damage (MtD) represents a dramatic change in cellular homeostasis, necessitating metabolic changes and stimulating mitophagy. One rapid response to MtD is a rapid peri-mitochondrial actin polymerization termed ADA (acute damage-induced actin). The activation mechanism for ADA is unknown. Here, we use mitochondrial depolarization or the complex I inhibitor metformin to induce ADA. We show that two parallel signaling pathways are required for ADA. In one pathway, increased cytosolic calcium in turn activates PKC-β, Rac, WAVE regulatory complex, and Arp2/3 complex. In the other pathway, a drop in cellular ATP in turn activates AMPK (through LKB1), Cdc42, and FMNL formins. We also identify putative guanine nucleotide exchange factors for Rac and Cdc42, Trio and Fgd1, respectively, whose phosphorylation states increase upon mitochondrial depolarization and whose suppression inhibits ADA. The depolarization-induced calcium increase is dependent on the mitochondrial sodium-calcium exchanger NCLX, suggesting initial mitochondrial calcium efflux. We also show that ADA inhibition results in enhanced mitochondrial shape changes upon mitochondrial depolarization, suggesting that ADA inhibits these shape changes. These depolarization-induced shape changes are not fragmentation but a circularization of the inner mitochondrial membrane, which is dependent on the inner mitochondrial membrane protease Oma1. ADA inhibition increases the proteolytic processing of an Oma1 substrate, the dynamin GTPase Opa1. These results show that ADA requires the combined action of the Arp2/3 complex and formin proteins to polymerize a network of actin filaments around mitochondria and that the ADA network inhibits the rapid mitochondrial shape changes that occur upon mitochondrial depolarization.

Vimentin intermediate filaments and filamentous actin form unexpected interpenetrating networks that redefine the cell cortex.

The cytoskeleton of eukaryotic cells is primarily composed of networks of filamentous proteins, F-actin, microtubules, and intermediate filaments. Interactions among the cytoskeletal components are important in determining cell structure and in regulating cell functions. For example, F-actin and microtubules work together to control cell shape and polarity, while the subcellular organization and transport of vimentin intermediate filament (VIF) networks depend on their interactions with microtubules. However, it is generally thought that F-actin and VIFs form two coexisting but separate networks that are independent due to observed differences in their spatial distribution and functions. In this paper, we present a closer investigation of both the structural and functional interplay between the F-actin and VIF cytoskeletal networks. We characterize the structure of VIFs and F-actin networks within the cell cortex using structured illumination microscopy and cryo-electron tomography. We find that VIFs and F-actin form an interpenetrating network (IPN) with interactions at multiple length scales, and VIFs are integral components of F-actin stress fibers. From measurements of recovery of cell contractility after transient stretching, we find that the IPN structure results in enhanced contractile forces and contributes to cell resilience. Studies of reconstituted networks and dynamic measurements in cells suggest direct and specific associations between VIFs and F-actin. From these results, we conclude that VIFs and F-actin work synergistically, both in their structure and in their function. These results profoundly alter our understanding of the contributions of the components of the cytoskeleton, particularly the interactions between intermediate filaments and F-actin.

MeerKAT view of the diffuse radio sources in Abell 3667 and their interactions with the thermal plasma

Context. During their lifetimes, galaxy clusters grow through the accretion of matter from the filaments of the large-scale structure and from mergers with other clusters. These mergers release a large amount of energy into the intracluster medium (ICM) through merger shocks and turbulence. These phenomena are associated with the formation of radio sources known as radio relics and radio halos, respectively. Radio relics and halos are unique proxies for studying the complex properties of these dynamically active regions of clusters and the microphysics of the ICM more generally. Aims. Abell 3667 is a spectacular example of a merging system that hosts a large pair of radio relics. Due to its proximity (z = 0.0553) and large mass, the system enables the study of these sources to a uniquely high level of detail. However, being located at Dec = −56.8°, the cluster could only be observed with a limited number of radio facilities. Methods. We observed Abell 3667 with MeerKAT as part of the MeerKAT Galaxy Cluster Legacy Survey. We used these data to study the large-scale emission of the cluster, including its polarisation and spectral properties. The results were then compared with simulations. Results. We present the most detailed view of the radio relic system in Abell 3667 to date, with a resolution reaching 3 kpc. The relics are filled with a network of filaments with different spectral and polarisation properties that are likely associated with multiple regions of particle acceleration and local enhancements of the magnetic field. Conversely, the magnetic field in the space between filaments has strengths close to what would be expected in unperturbed regions at the same cluster-centric distance. Comparisons with magnetohydrodynamic cosmological and Lagrangian simulations support the idea of filaments as multiple acceleration sites. Our observations also confirm the presence of an elongated radio halo, developed in the wake of the bullet-like sub-cluster that merged from the south-east. Finally, we associate the process of magnetic draping with a thin polarised radio source surrounding the remnant of the bullet’s cool core. Conclusions. Our observations have unveiled the complexity of the interplay between the thermal and non-thermal components in the most active regions of a merging cluster. Both the intricate internal structure of radio relics and the direct detection of magnetic draping around the merging bullet are powerful examples of the non-trivial magnetic properties of the ICM. Thanks to its sensitivity to polarised radiation, MeerKAT will be transformational in the study of these complex phenomena.

Interplay between Brownian motion and cross-linking controls bundling dynamics in actin networks

Morphology changes in cross-linked actin networks are important in cell motility, division, and cargo transport. Here, we study the transition from a weakly cross-linked network of actin filaments to a heavily cross-linked network of actin bundles through microscopic Brownian dynamics simulations. We show that this transition occurs in two stages: first, a composite bundle network of small and highly aligned bundles evolves from cross-linking of individual filaments and, second, small bundles coalesce into the clustered bundle state. We demonstrate that Brownian motion speeds up the first stage of this process at a faster rate than the second. We quantify the time to reach the composite bundle state and show that it strongly increases as the mesh size increases only when the concentration of cross-links is small and that it remains roughly constant if we decrease the relative ratio of cross-linkers as we increase the actin concentration. Finally, we examine the dependence of the bundling timescale on filament length, finding that shorter filaments bundle faster because they diffuse faster.

Gelsolin impairs barrier function in pancreatic ductal epithelial cells by actin filament depolymerization in hypertriglyceridemia-induced pancreatitis in vitro.

Gelsolin (GSN) is a calcium-regulated actin-binding protein that can sever actin filaments. Notably, actin dynamics affect the structure and function of epithelial barriers. The present study investigated the role of GSN in the barrier function of pancreatic ductal epithelial cells (PDECs) in hypertriglyceridemia-induced pancreatitis (HTGP). The human PDEC cell line HPDE6-C7 underwent GSN knockdown and was treated with caerulein (CAE) + triglycerides (TG). Intracellular calcium levels and the actin filament network were analyzed under a fluorescence microscope. The expression levels of GSN, E-cadherin, nectin-2, ZO-1 and occludin were evaluated by reverse transcription-quantitative polymerase chain reaction and western blotting. Ultrastructural changes in tight junctions were observed by transmission electron microscopy. Furthermore, the permeability of PDECs was analyzed by fluorescein isothiocyanate-dextran fluorescence. The results revealed that CAE + TG increased intracellular calcium levels, actin filament depolymerization and GSN expression, and increased PDEC permeability by decreasing the expression levels of E-cadherin, nectin-2, ZO-1 and occludin compared with the control. Moreover, changes in these markers, with the exception of intracellular calcium levels, were reversed by silencing GSN. In conclusion, GSN may disrupt barrier function in PDECs by causing actin filament depolymerization in HTGP in vitro.