Search Results (432)

Background/Objectives: Zebrafish fin regeneration serves as a classic model for investigating vertebrate tissue regeneration, yet the core regulatory networks and their crosstalk with the immune microenvironment remain incompletely characterized. This study aimed to identify hub genes, and elucidate the underlying molecular mechanisms and immune microenvironment dynamics during zebrafish fin regeneration. Methods: We integrated multiple bulk RNA-seq datasets of zebrafish fin regeneration from the GEO database, followed by data standardization with batch effect removal. Hub genes were screened via differential expression analysis, weighted gene co-expression network analysis (WGCNA), and predictive models constructed with 13 classic machine learning algorithms. Functional enrichment, time-ordered gene co-expression network (TO-GCN) method, immune infiltration analyses and RT-qPCR validation were further performed. Results: We identified upregulated differentially expressed genes, regeneration-correlated gene modules and their overlapping genes, including 82 candidate genes and 10 hub genes enriched in cytoskeleton remodeling, extracellular matrix organization, and focal adhesion. Temporal analysis uncovered hierarchical gene regulation and functional switching during regeneration. Hub gene expression was significantly correlated with the infiltration of B cells, M1/M2 macrophages and CD8+ T cells, revealing a stage-specific immune microenvironment. RT-qPCR validation showed high consistency with the multi-omics data. Conclusions: This study provides potential gene targets for understanding zebrafish fin regeneration, and offers a valuable reference for investigating the crosstalk between regulatory networks and the immune microenvironment in vertebrate tissue regeneration. Full article

Soil salinity is a major abiotic stressor that constrains plant growth and development, yet the coordinated regulatory mechanisms underlying salt stress impacts on plant cell mechanical properties and the cytoskeleton remain elusive. In this study, tobacco suspension cells were employed as a model system. Combining mechanical measurements, fluorescence microscopy imaging, and bright-field morphological observation, we systematically characterized the dynamic response patterns of cell-generated surface stress (ΔS), cell viscoelastic index (CVI), microfilament cytoskeleton structure, as well as cell morphology and plasmolysis under NaCl stress ranging from 50 to 150 mmol/L. The results revealed three distinct response thresholds: 50 mmol/L NaCl treatment induced only transient ΔS fluctuations and mild plasmolysis, with no significant changes in CVI or microfilament fluorescence intensity, suggesting a safe tolerance threshold. The 75–100 mmol/L NaCl treatments triggered reversible “rise–recovery” mechanical responses in ΔS and CVI. The microfilament cytoskeleton showed minor structural adjustments, and plasmolysis increased gradually but remained reversible, defining this range as a reversible acclimation phase. The 125–150 mmol/L NaCl treatment caused an irreversible decline in ΔS (with a sharp instantaneous drop at 150 mmol/L). CVI variations diminished and stabilized after 6 h. The microfilament cytoskeleton suffered progressive disruption, as fluorescence intensity dropped to 1% of the control group at 150 mmol/L, accompanied by severe plasmolysis and protoplast shrinkage, indicating irreversible cellular damage. These findings demonstrate a concentration-dependent gradient effect of NaCl stress, highlighting tight coordination between mechanical properties, cytoskeletal integrity, and morphological adaptation. This work provides critical cytological insights into the molecular regulation of plant salt stress responses. Full article

Titanium and its alloys are widely used for orthopedic implants, but their intrinsic bioinertness may hinder osseointegration. In this study, titanium dioxide nanotube (TNT) arrays were fabricated on Ti-6Al-4V scaffolds via anodization, and their effects on the adhesion behavior of human bone marrow mesenchymal stem cells (hBMSCs) were investigated. Surface characterization showed that anodization successfully generated ordered TNT layers, increased surface roughness, enhanced protein adsorption, and induced an apparent superhydrophilic wetting response. Compared to the untreated scaffold and TNT50, the small-diameter TNT10 surface significantly promoted hBMSC adhesion and proliferation. Microscope imaging further revealed enhanced cell spreading, F-actin organization, and vinculin expression on TNT surfaces, with the most prominent focal adhesion-related staining observed in TNT10. Quantitative proteomic analysis showed that TNT10 was associated with coordinated remodeling of adhesion- and cytoskeleton-related molecular programs, including focal adhesion, cell–substrate junction, and regulation of the actin cytoskeleton. In contrast, TNT50, despite supporting obvious cytoskeletal remodeling, was more compatible with a dynamic, higher-turnover adhesion state. Overall, these findings suggest that small-diameter TNTs provide a more favorable interfacial microenvironment for stable early hBMSC adhesion on porous titanium scaffolds. Full article

Eating behavior requires a balance between metabolic and hedonic components. Anxiety and dietary type may influence the quantity, patterns, and other aspects of food intake. Modern diets, especially in Western societies, often contain high levels of calories from fat and simple sugars (e.g., cafeteria-style diets). This type of diet may promote overweight and/or obesity in some, although many consumers remain at a normal weight. The mechanisms underlying susceptibility or resistance to weight gain remain unclear. Here, Sprague-Dawley male rats were fed a cafeteria diet for 10 weeks and then classified into quartiles based on body mass. We evaluated locomotor activity and anxiety-like behaviors and analyzed hypothalamic proteomics in overweight (Q4) rats compared with underweight (Q1) rats. Our results showed that locomotor activity and anxiety-like behaviors did not differ across quartiles (p > 0.05). Nevertheless, the expression of several hypothalamic proteins differed between Q4 and Q1 rats. Functional enrichment analysis of these differentially expressed proteins (p ≤ 0.05) revealed changes in cytoskeleton dynamics, synaptic communication, energy production and utilization, biosynthesis of cellular components (including nucleotides and carbohydrates), and regulation of metabolism between Q1 and Q4 rats. Neuro-humoral hypothalamic output regulates metabolism and food intake. Therefore, these functional changes in the hypothalamus may be associated with rats’ susceptibility/resistance to weight gain. Full article

Actin is a key component of the cytoskeleton and plays diverse roles in cellular processes. Autophagy regulates homeostasis through various mechanisms that recycle nutrients and degrade unnecessary or harmful cellular components and aggregates. These two processes are engaged in a highly conserved crosstalk through which they regulate each other, including autophagolysosomal formation and regulation of actin dynamics. The regulation of autophagy is involved in cancer, neurodegeneration, infectious diseases, and inflammation, providing possible avenues for treatments for these diseases. In this review, we summarize current knowledge of the actin–autophagy interplay and regulation, and explore the possible implications for disease progression and therapies. Although more research is necessary to strengthen the effectiveness of therapies that target the regulation of autophagy and actin dynamics, significant strides have already been made, clearly indicating the potential benefit of targeting these processes. Full article

The fall armyworm (Spodoptera frugiperda) is a major global migratory pest. Its increasing insecticide resistance poses a severe threat to food security. Developing biopesticides such as SfMNPV is critical for sustainable control. Nevertheless, the early molecular mechanisms underlying the S. frugiperda midgut response to oral SfMNPV challenge remain poorly understood. This study utilized high-throughput transcriptome sequencing to systematically characterize the dynamic transcriptional profiles of the larval midgut at 1, 12, and 24 h after oral SfMNPV inoculation. Results showed that the midgut transcriptional response to SfMNPV is time and stage-specific. During this period, the physical midgut barrier underwent remodeling, with core components of the peritrophic matrix downregulated at 1 h, followed by the basal lamina at 12 h, alongside the activation of cytoskeleton genes during 12–24 h. Concurrently, sustained endoplasmic reticulum stress, autophagy, and ubiquitin system responses occurred from 12 to 24 h. At the metabolic level, the defense system exhibited a functional succession, shifting from ABC transporters and UDP-glycosyltransferases at 1 h to glutathione S-transferases and superoxide dismutase at 12–24 h. Additionally, the midgut tissue exhibited a cascade transition from pro-apoptotic signaling at 1 h to compensatory regenerative repair mediated by the Wnt, mTOR, and Hippo pathways at 12–24 h. This study elucidates the molecular process of barrier damage, homeostatic imbalance, and tissue remodeling during early oral SfMNPV challenge. These findings provide a global perspective on baculovirus-host interactions and establish a theoretical foundation for designing novel biopesticides targeting the midgut interaction. Full article

Background/Objectives. The actin-binding protein EPLIN (epithelial protein lost in neoplasm), also known as LIMA1, contributes to the maintenance of cytoskeleton structure and dynamic. This protein, which interacts with multiple partners to regulate cell adhesion and migration, has been implicated in the progression of solid tumors and in tumor metastasis. Consequently, small molecules binding to EPLIN are actively searched. EPLIN has been characterized as a molecular target for the antitumor antibiotic albacarcin V which affects the cytoskeletal structure and induces cell growth arrest. Methods. We have modeled the binding of albacarcin and naturally occurring derivatives to EPLIN conformers, in order to locate the drug-binding site and to identify additional EPLIN binders. Nineteen compounds were studied, including albacarcins V (vinyl) and M (methyl), five gilvocarcins, four ravidomycins, two chrysomycins, and six related products (including polycarcin and fucomycin). Results. The modeling analysis confirmed the capacity of albacarcin V to bind to EPLIN and identified a few better binders. In particular, ravidomycin V bearing a dimethylamino sugar unit were identified as the best binders in the series, along with the two related anticancer natural products FE35A-B. Structure-binding relationships are discussed. The drug-binding site has been localized near the central residue Asn34 in the conformationally constrained domain between the two zinc-binding regions. Conclusions. This study provides guidance to the design of EPLIN inhibitors based on the ravidomycin core structure. Full article

Background/Objectives: Among cotton species, Gossypium barbadense produces the strongest fibers. Examining cytoskeletal dynamics in single epidermal cells of G. barbadense ovules offers a direct approach to investigating fiber quality. Microtubules are major cytoskeletal components whose organization and dynamics are precisely regulated by microtubule-associated proteins (MAPs). However, information on the TPX2 family remains limited, and characterizing its features in G. barbadense is critical to clarifying the role of TPX2 family members in fiber strength formation. Methods: Using the Arabidopsis thaliana TPX2 sequence as a reference, 40, 49, 26, and 26 TPX2 family members were identified in the genomes of G. barbadense, Gossypium hirsutum, Gossypium arboreum, and Gossypium raimondii, respectively. We further analyzed the expression pattern of GbTPX2-35 and validated its function via virus-induced gene silencing (VIGS). Results: In G. barbadense, GbTPX2-35 (Gbar_D11G59825.1) was significantly upregulated in fiber samples of the parental lines at 25 days post-anthesis, and this expression pattern was further validated in G. barbadense lines with extreme fiber strength phenotypes. Next, VIGS-mediated silencing of GbTPX2-35 downregulated the transcript levels of cellulose synthase and microtubule-related protein genes, a finding further validated by mature fiber strength phenotypic data. Conclusions: This study preliminarily validated a pathway in which GbTPX2-35 regulates fiber strength by coordinating cellulose biosynthesis with microtubule cytoskeleton dynamics, providing valuable candidate genes and theoretical support for molecular breeding of high-strength cotton fibers. Full article

Nuclear mechanics and mechanotransduction are involved in the migration and invasion process, such as those in which the cells need to deform themselves to pass through constrictions. Specifically, properties like nuclear softness, viscoelasticity, plasticity (like nuclear pore complexes) and deformability are critical in cancer and its malignant progression. The nucleus represents a physical barrier for the migration and invasion in dense 3D extracellular matrix (ECM) scaffolds. Therefore, the deformability of the nucleus seems to determine the migration limit in circumstances where the enzymatic remodeling of the surroundings is impaired. There are still significant knowledge gaps regarding effects of nuclear deformation during cancer dissemination. It seems that nuclear deformation can alter gene transcription, induce alternative splicing processes, impact nuclear envelope rupture, nuclear pore complex dilatation, damage the DNA, and increase the genomic instability. These mechanically induced alterations can in turn impact the migratory behavior of the cancer cells. The stiffness of the nucleus relies on the condensation of chromatin, and the nuclear lamina, which consists of a network of intermediate filaments underneath the nuclear envelope. All of this is discussed in the review and it is argued that nuclear deformability is universally found in various cancer types. Another focus is placed on the nuclear envelope proteins like emerin, and the SUN-KASH complex and how they contribute to the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex, which consequently couples the nucleus and the cytoskeleton. It is argued that this connection is crucial for force transmission, which governs nuclear stiffness dynamically, depending on the force applied. In this review, recent findings are described that couple ECM-induced nuclear mechanosensing and mechanotransduction with the migration and invasion of cancer cells. Moreover, it is suspected that changes in the mechanosensory characteristics of the cell nucleus could play a pivotal part in the malignancy of cancer cells and the heterogeneity of tumors. Finally, it is discussed what impact the individual elements of the nucleus offer to mechanically alter cellular migration and invasion in cancer and its malignant progression. Full article

Neuronal function relies on the precise coordination between intracellular calcium (Ca²⁺) signaling and the cytoskeletal architecture that underpins synaptic transmission, plasticity, and structural stability. Disruption of this calcium–cytoskeleton interplay has been noted in numerous neurodegenerative diseases. We discuss how Ca²⁺-dependent cytoskeletal remodeling governs long-term potentiation and depression, dendritic spine morphology, and presynaptic function, highlighting the functions of end-binding proteins, STIM (Stromal Interaction Molecule)/Orai-mediated store-operated calcium entry, and the spine apparatus. Disease-specific manifestations of cytoskeletal–calcium dysregulation are reviewed across Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, tauopathies, and prion disorders. Finally, we evaluate emerging therapeutic strategies targeting calcium homeostasis, cytoskeletal dynamics, and their downstream effectors, including multi-target approaches. Full article

Organ-on-a-chip devices combine microfabrication, tissue engineering, and microfluidics to recreate physiologically relevant microenvironments for in vitro studies. In this work, we validated a dynamic 2D breast-on-a-chip microfluidic bioassay operated at a controlled infusion rate of 20 µL/h to assess anticancer drug responses under defined flow conditions. Using Palbociclib as a reference compound, we evaluated proliferation, viability/apoptosis, cytoskeleton organization, and differential processing of the resistance-associated marker PARP1 in MCF-7 and T47D breast cancer cells. Under dynamic microfluidic conditions, Palbociclib induced dose-dependent effects, with the higher concentration (20 µM) consistently reducing cell proliferation and viability and increasing late apoptosis compared to 10 µM Palbociclib. Cytoskeletal disorganization was observed at both concentrations, while differential PARP1 processing patterns between MCF-7 and T47D cells were detected across doses. These responses are consistent with known effects of CDK4/6 inhibition and were reproducibly captured under controlled flow conditions. Overall, our results demonstrate that this breast-on-a-chip microfluidic model provides a reproducible and physiologically relevant in vitro platform for integrated assessment of drug efficacy and resistance-associated markers under dynamic perfusion. Full article

Electrophysiology, mechanobiology, and the study of soft matter within cells demonstrate increasing amounts of evidence that neuronal signaling arises from interactions between membrane potential, force, and phase. Herein, we have attempted to collect and organize the evidence for each of these areas of study into an approximate structure called the electromechanical connectome: a three-way state–space (membrane potentials, nanoscale mechanical forces, and cytoplasmic rheology, including phase-separated liquid–liquid droplets) where membrane potentials, nanoscale mechanical forces, and cytoplasmic rheology, and phase-separated liquid–liquid droplets are likely to influence one another, influencing synaptic processing, plasticity and network stability. We will also attempt to illustrate the following: how changes in electrostatic fields can be used to alter the arrangement of lipids, hydration, and dielectric microdomains, and the contact geometry between organelles and activity dependent transcription; how mechanical dynamics associated with spines, axons, and the active zone of synapses may be used to modify the energy landscape of channels, the docking and priming of vesicles, and the transport of cytoskeletons; and how viscosity corridors, along with phase-separated micro-reactors, can be used to regulate the kinetics of signaling, molecular trafficking and metabolic processes in local environments. With these connections in mind, we will propose a multiphysical attractor model in which cognition is the result of navigating through metastable manifolds, while neurodegenerative disease may be a result of the progressive loss of electromechanical coherence, phase boundary control and energetic flexibility. Finally, we will present testable hypotheses and use AI-enabled digital twin methods to potentially quantify the early deformation of manifolds and provide precision biomarkers and therapeutic options. Full article

Metastatic breast cancer (BC) remains a major clinical challenge, and identifying molecular mechanisms driving tumor cell migration and invasion is critical to develop effective therapeutic strategies. Clusterin (CLU), a secreted chaperone-like protein, is upregulated in BC and metastatic tissue; however, its functional contribution to tumor aggressiveness remains unclear. Here, we silenced CLU by siRNA in two BC cell lines with distinct aggressiveness and examined its impact on migration, invasion, and associated signaling pathways. Following CLU silencing, cell migration and invasion were assessed using transwell assays. Cytoskeletal organization was evaluated by F-actin staining, while downstream signaling pathways were analyzed by RT-PCR, Western blotting, and Rho GTPase pull-down. A comparative proteomic analysis was performed in CLU-expressing and CLU-silenced MDA-MB-231 cells. CLU knockdown significantly reduced migration and invasion in MDA-MB-231, concomitantly with loss of F-actin-rich membrane protrusions, reduced expression of MMP9, COL1A1, and COL4A1, and decreased activation of Akt, NF-κB, and RhoA. Proteomic profiling revealed extensive remodeling of pathways involved in cell adhesion, cytoskeletal dynamics, and extracellular matrix interactions. Differently, no or very mild effects were observed in CLU-silenced MCF-7 cells. These findings identify CLU as an upstream regulator of a pro-metastatic adhesion–cytoskeleton signaling in BC, selectively operative in EMT-engaged, basal-like cells, highlighting the importance of patient stratification for CLU-targeted therapeutic strategies. Full article

Alzheimer’s disease (AD) is characterized by progressive cognitive decline, with amyloid beta oligomers (AβOs) emerging as the most neurotoxic species and acting as early triggers of cellular alterations. Before the appearance of other protein aggregates, AβOs disrupt the dynamics and stability of the neuronal cytoskeleton, a structure essential for maintaining neuronal morphology, axonal transport, and synaptic plasticity. Experimental evidence demonstrates that AβOs promote microtubule disassembly, Tau hyperphosphorylation, reduced kinesin levels, impaired axonal transport, and alterations in actin dynamics through the LIMK–cofilin signaling pathway. In addition, increased levels of neurofilament light chain have been identified as an early biomarker of axonal damage. Notably, these cytoskeletal disturbances arise in the absence of extensive neuronal death, underscoring the cytoskeleton as a critical early target in AD pathogenesis. In this review, we analyze cytoskeletal alterations induced by AβOs in neurons and discuss how these changes may contribute to disrupted neuronal communication, a defining early hallmark of AD pathology. Full article