Actin filaments assemble into a variety of networks to provide force for diverse cellular processes 1. Tropomyosins are coiled-coil dimers that form head-to-tail polymers along actin filaments and ...regulate interactions of other proteins, including actin-depolymerizing factor (ADF)/cofilins and myosins, with actin 2–5. In mammals, >40 tropomyosin isoforms can be generated through alternative splicing from four tropomyosin genes. Different isoforms display non-redundant functions and partially non-overlapping localization patterns, for example within the stress fiber network 6, 7. Based on cell biological studies, it was thus proposed that tropomyosin isoforms may specify the functional properties of different actin filament populations 2. To test this hypothesis, we analyzed the properties of actin filaments decorated by stress-fiber-associated tropomyosins (Tpm1.6, Tpm1.7, Tpm2.1, Tpm3.1, Tpm3.2, and Tpm4.2). These proteins bound F-actin with high affinity and competed with α-actinin for actin filament binding. Importantly, total internal reflection fluorescence (TIRF) microscopy of fluorescently tagged proteins revealed that most tropomyosin isoforms cannot co-polymerize with each other on actin filaments. These isoforms also bind actin with different dynamics, which correlate with their effects on actin-binding proteins. The long isoforms Tpm1.6 and Tpm1.7 displayed stable interactions with actin filaments and protected filaments from ADF/cofilin-mediated disassembly, but did not activate non-muscle myosin IIa (NMIIa). In contrast, the short isoforms Tpm3.1, Tpm3.2, and Tpm4.2 displayed rapid dynamics on actin filaments and stimulated the ATPase activity of NMIIa, but did not efficiently protect filaments from ADF/cofilin. Together, these data provide experimental evidence that tropomyosin isoforms segregate to different actin filaments and specify functional properties of distinct actin filament populations.
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•Stress-fiber-associated tropomyosin isoforms segregate to different actin filaments•Tropomyosin isoforms bind F-actin with different dynamics•Dynamic tropomyosin isoforms activate non-muscle myosin II•Stable tropomyosin isoforms protect actin filaments from ADF/cofilin
Gateva et al. report that distinct tropomyosin isoforms segregate to different actin filaments and can specify functional properties of distinct actin filament populations. They also provide evidence that functions of tropomyosins in myosin II activation and actin filament stabilization correlate with the dynamics of their actin interactions.
Although the neural systems supporting interoception have been outlined in general, the exact processes underlying the integration of visceral signals still await research. Based on the predictive ...coding concept, we aimed to reveal the neural networks responsible for the bottom-up (stimulus-dependent) and top-down (model-dependent) processing of interoceptive information. In a study of 30 female participants, we utilized two classical body perception experiments—the rubber hand illusion and a heartbeat detection task (cardioception), with the latter being implemented in fMRI settings. We interpreted a stronger rubber hand illusion, as measured by higher proprioceptive drift, as a tendency to rely on actual sensory experience, i.e., bottom-up processing, while lower proprioceptive drift served as an indicator of the prevalence of top-down, model-based influences. To reveal the bottom-up and top–down processes in cardioception, we performed a seed-based connectivity analysis of the heartbeat detection task, using as seeds the areas with known roles in sensory integration and entering proprioceptive drift as a covariate. The results revealed a left thalamus-dependent network positively associated with proprioceptive drift (bottom-up processing) and a left amygdala-dependent network negatively associated with drift (top-down processing). Bottom-up processing was related to thalamic connectivity with the left frontal operculum and anterior insula, anterior cingulate cortex, hypothalamus, right planum polare and right inferior frontal gyrus. Top-down processing was related to amygdalar connectivity with the rostral prefrontal cortex and an area involving the left frontal opercular and anterior insular cortex, with the latter area being an intersection of the two networks. Thus, we revealed the neural mechanisms underlying the integration of interoceptive information through the interaction between the current sensory experience and internal models.
Cerebral small vessel disease (CSVD) is a significant cause of cognitive impairment (CI), disability, and mortality. The insufficient effectiveness of antihypertensive therapy in curbing the disease ...justifies the search for potential targets for modifying therapy and indicators supporting its use. Using a laser-assisted optical rotational cell analyzer (LORRCA, Mechatronics, The Netherlands), the rheological properties and deformability of erythrocytes before and after incubation with 10 μmol/L of L-arginine, the nitric oxide (NO) donor, blood–brain barrier (BBB) permeability assessed by dynamic contrast-enhanced MRI, clinical, and MRI signs were studied in 73 patients with CSVD (48 women, mean age 60.1 ± 6.5 years). The control group consisted of 19 volunteers (14 women (73.7%), mean age 56.9 ± 6.4 years). The erythrocyte disaggregation rate (y-dis) after incubation with L-arginine showed better performance than other rheological characteristics in differentiating patients with reduced NO bioavailability/NO deficiency by its threshold values. Patients with y-dis > 113 s−1 had more severe CI, arterial hypertension, white matter lesions, and increased BBB permeability in grey matter and normal-appearing white matter (NAWM). A test to assess changes in the erythrocyte disaggregation rate after incubation with L-arginine can be used to identify patients with impaired NO bioavailability. L-arginine may be part of a therapeutic strategy for CSVD with CI.
Cerebral small vessel disease (SVD) is a major cause of cognitive impairment in elderly people. While most research focuses on the role of the classical vascular risk factors in SVD, a description of ...the psychophysiological mechanisms leading to the age‐related brain damage may open new possibilities for prophylaxis. In the current study, we evaluated the associations between emotional abilities, interoception, and age‐related vascular white matter degeneration. The work was influenced, first, by multiple studies recognizing alexithymia as a cardiovascular risk factor; second, by theories of emotions linking body's allostasis and emotional regulation; and third, by neuroimaging data highlighting the shared role of the insular cortex in interoceptive and emotional processing. In a sample of older female adults (N = 30), we performed the Mayer–Salovey–Caruso Emotional Intelligence Test, functional MRI using the heartbeat detection task, and evaluation of white matter microstructural integrity using diffusion weighted imaging. The ability to understand and analyze emotions—one of the four components of emotional intelligence—was found to be associated with higher interoception‐related activation of the right anterior insula and preserved white matter microstructure. We interpret these results in light of the concept of Embodied Predictive Interoception Coding, which proposes that emotional processing, interoception, and allostasis (antecedent top‐down regulation of the body's internal milieu) may rely on the shared neural mechanisms of predictive coding. The study demonstrates feasibility of the investigation of cerebrovascular diseases form a psychophysiological perspective and calls for future research.
Associations between emotional intelligence and age‐related cerebrovascular brain injury are revealed in the context of neuroscientific theories of emotion and interoception, using functional neuroimaging with the heartbeat detection task. We have shown that the ability to understand emotions is related to higher insular activation and to preserved white matter microstructure.
ADF/cofilins drive cytoskeletal dynamics by promoting the disassembly of “aged” ADP-actin filaments. Mammals express several ADF/cofilin isoforms, but their specific biochemical activities and ...cellular functions have not been studied in detail. Here, we demonstrate that the muscle-specific isoform cofilin-2 promotes actin filament disassembly in sarcomeres to control the precise length of thin filaments in the contractile apparatus. In contrast to other isoforms, cofilin-2 efficiently binds and disassembles both ADP- and ATP/ADP-Pi-actin filaments. We mapped surface-exposed cofilin-2-specific residues required for ATP-actin binding and propose that these residues function as an “actin nucleotide-state sensor” among ADF/cofilins. The results suggest that cofilin-2 evolved specific biochemical and cellular properties that allow it to control actin dynamics in sarcomeres, where filament pointed ends may contain a mixture of ADP- and ATP/ADP-Pi-actin subunits. Our findings also offer a rationale for why cofilin-2 mutations in humans lead to myopathies.
•Cofilin-2 promotes actin filament disassembly at the M-band of muscle sarcomere•Cofilin-2 is critical for regulation of precise thin filament length in sarcomeres•ADF/cofilins harbor a nucleotide state sensor that interacts with actin subdomain 3
Generation of contractile force in skeletal muscles relies on careful regulation of actin dynamics. Kremneva et al. identify unique features of cofilin-2, a muscle-specific isoform of the actin binding protein, that allow it to efficiently disassemble both ATP- and ADP-Pi-actin filaments to regulate their length in cardiac muscle sarcomeres.
Age-dependent cerebral small vessel disease (CSVD) is a common disease with a high social burden characterized by heterogeneity of forms and frequent comorbidity with Alzheimer’s disease (AD). ...Previously, we identified two MRI types of CSVD with specific clinical presentation and, probably, different mechanisms. The present study included 34 patients with CSVD and white matter hyperintensity (WMH) of stage Fazekas (F) 3 (mean age 61.7 ± 8.9) and 11 volunteers (mean age 57.3 ± 9.7). Total RNA was isolated from peripheral blood leukocytes. The expression of 58 protein-coding genes associated with CSVD and/or AD and 4 reference genes were assessed as part of the original panel for the NanoString nCounter analyzer. Testing results were validated by real-time PCR. There was a significant decrease in the expression levels of the ACOX1, CD33, CD2AP, TNFR1, and VEGFC genes in MRI type 2 relative to the control group as well as a decrease in the expression level of the CD33 gene in MRI type 2 compared to MRI type 1. Processes associated with inflammatory pathways with decreased expression of the identified genes are important in the development of MRI type 2 of CSVD. Given the direct connection of the established genes with AD, the importance of this form of CSVD in comorbidity with AD has been assumed.
Aging is known to be associated with a decline in interoceptive abilities and changes in emotional processing, including alexithymia. As the brain areas supporting interoceptive awareness participate ...in the perception of emotion, we suggested that interoceptive decline and alexithymia in older adults may share common neural ground. To test this hypothesis, we administered functional magnetic resonance imaging-based heartbeat detection task to 62 adults of diverse ages (range 18-73) and evaluated a larger sample of older and younger adults using questionnaires characterizing interoceptive sensibility, alexithymia, and depressive attitudes. We found that increasing age was linked to decreased activation during the interoceptive task, including the right insular-opercular and supplementary motor areas (SMAs). Age also affected task-based functional connectivity, with two major effects being a decrease in the connectivity of the SMA-insular network and an increase in the connectivity of the prefrontal-lateral occipital network. Path analysis performed for interoceptive accuracy as the endogenous variable revealed that the impact of age was mediated by the functional activation of the insular cortex and SMA and by the connectivity between these areas. Another path analysis using alexithymia as the endogenous variable while controlling for depressive attitudes showed that the effect of age was mediated by interoceptive decline. The study supports the role of central mechanisms in age-related interoceptive decline and shows its implications for alexithymia. Since alexithymia represents a risk factor for mental and cardiovascular diseases, the study findings may open an important direction toward maintaining older adults' well-being. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
Focal adhesions (FAs) connect inner workings of cell to the extracellular matrix to control cell adhesion, migration and mechanosensing. Previous studies demonstrated that FAs contain three vertical ...layers, which connect extracellular matrix to the cytoskeleton. By using super-resolution iPALM microscopy, we identify two additional nanoscale layers within FAs, specified by actin filaments bound to tropomyosin isoforms Tpm1.6 and Tpm3.2. The Tpm1.6-actin filaments, beneath the previously identified α-actinin cross-linked actin filaments, appear critical for adhesion maturation and controlled cell motility, whereas the adjacent Tpm3.2-actin filament layer beneath seems to facilitate adhesion disassembly. Mechanistically, Tpm3.2 stabilizes ACF-7/MACF1 and KANK-family proteins at adhesions, and hence targets microtubule plus-ends to FAs to catalyse their disassembly. Tpm3.2 depletion leads to disorganized microtubule network, abnormally stable FAs, and defects in tail retraction during migration. Thus, FAs are composed of distinct actin filament layers, and each may have specific roles in coupling adhesions to the cytoskeleton, or in controlling adhesion dynamics.
Sharpin, a multifunctional adaptor protein, regulates several signalling pathways. For example, Sharpin enhances signal-induced NF-κB signalling as part of the linear ubiquitin assembly complex ...(LUBAC) and inhibits integrins, the T cell receptor, caspase 1 and PTEN. However, despite recent insights into Sharpin and LUBAC function, a systematic approach to identify the signalling pathways regulated by Sharpin has not been reported. Here, we present the first 'Sharpin interactome', which identifies a large number of novel potential Sharpin interactors in addition to several known ones. These data suggest that Sharpin and LUBAC might regulate a larger number of biological processes than previously identified, such as endosomal trafficking, RNA processing, metabolism and cytoskeleton regulation. Importantly, using the Sharpin interactome, we have identified a novel role for Sharpin in lamellipodium formation. We demonstrate that Sharpin interacts with Arp2/3, a protein complex that catalyses actin filament branching. We have identified the Arp2/3-binding site in Sharpin and demonstrate using a specific Arp2/3-binding deficient mutant that the Sharpin-Arp2/3 interaction promotes lamellipodium formation in a LUBAC-independent fashion.This article has an associated First Person interview with the first author of the paper.
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