Calcium waves facilitate and coordinate the contraction of endfeet actin stress fibers in Drosophila interommatidial cells

Actomyosin contraction shapes the Drosophila eye’s panoramic view. The convex curvature of the retinal epithelium, organized in ∼800 close-packed ommatidia, depends upon a fourfold condensation of the retinal floor mediated by contraction of actin stress fibers in the endfeet of interommatidial cells (IOCs). How these tensile forces are coordinated is not known. Here, we discover a novel phenomenon: Ca2+ waves regularly propagate across the IOC network in pupal and adult eyes. Genetic evidence demonstrates that IOC Ca2+ waves are independent of phototransduction, but require inositol 1,4,5-triphosphate receptor (IP3R), suggesting these waves are mediated by Ca2+ releases from ER stores. Removal of IP3R disrupts stress fibers in IOC endfeet and increases the basal retinal surface by ∼40%, linking IOC waves to facilitating stress fiber contraction and floor morphogenesis. Further, IP3R loss disrupts the organization of a collagen IV network underneath the IOC endfeet, implicating ECM and its interaction with stress fibers in eye morphogenesis. We propose that coordinated Ca2+ spikes in IOC waves promote stress fiber contractions, ensuring an organized application of the planar tensile forces that condense the retinal floor.Summary StatementCa2+ waves have an important role in generating tensile forces to shape the Drosophila eye’s convex curvature. Coordinated Ca2+ spikes facilitate actomyosin contractions at the basal endfeet of interommatidial cells.

The Formin Inhibitor, SMIFH2, Inhibits Members of the Myosin Superfamily

The small molecular inhibitor of formin FH2 domains, SMIFH2, is widely used in cell biological studies. It inhibits formin-driven actin polymerization in vitro, but not polymerization of pure actin. It is active against several types of formins from different species (Rizvi et al., 2009). Here, we found that SMIFH2 inhibits retrograde flow of myosin 2 filaments and contraction of stress fibers. We further checked the effect of SMIFH2 on non-muscle myosin 2A and skeletal muscle myosin 2 in vitro and found that SMIFH2 inhibits myosin ATPase activity and ability to translocate actin filaments in the in vitro motility assay. The inhibition of non-muscle myosin 2A in vitro required a higher concentration of SMIFH2 than for the inhibition of retrograde flow and stress fiber contraction in cells. We also found that SMIFH2 inhibits several other non-muscle myosin types, e.g. mammalian myosin 10, Drosophila myosin 7a and Drosophila myosin 5, more efficient than inhibition of formins. These off-target inhibitions demand additional careful analysis in each case when solely SMIFH2 is used to probe formin functions.

Contraction of muscle tissue during heat treatment

The results of the research established a link between the tension of the muscle fiber contraction and a temperature of heating. The obtained data indicate that the value of the tension of the contraction depends on the cross-sectional area of meat samples — the larger the cross-sectional area, the lower the tension of the contrac-tion, which is conditioned by non-uniform heating due to low thermal conductivity of meat. The equation characterizing the dependence of the tension of the muscle fiber contraction on the cross-sectional area of samples is presented. It was estab-lished that the value of the tension of the muscle contraction depended on meat raw material properties. The lowest value of the tension of the contraction was recorded in PSE meat, which, according to the authors’ opinion, was conditioned by abnormal autolysis and loss of functional properties by muscle proteins.

The Formin Inhibitor, SMIFH2, Inhibits Members of the Myosin Superfamily

The small molecular inhibitor of formin FH2 domains, SMIFH2, is widely used in cell biological studies. It was selected in a chemical screen as a compound inhibiting formin-driven actin polymerization in vitro, but not polymerization of pure actin, and found to be active against several types of formins from different species (Rizvi et al., 2009). Here, in experiments with cultured fibroblasts, we found that SMIFH2 inhibits retrograde flow of myosin 2 filaments and contraction of stress fibers. We further checked the effect of SMIFH2 on non-muscle myosin 2A and skeletal muscle myosin 2 in vitro and found that SMIFH2 inhibits myosin ATPase activity and ability to translocate actin filaments in the in vitro motility assay. While inhibition of myosin 2A in vitro required somewhat higher concentration of SMIFH2 than inhibition of retrograde flow and stress fiber contraction in cells, inhibition of several other non-muscle myosin types, e.g. mammalian myosin 10, Drosophila myosin 7a and Drosophila myosin 5 by SMIFH2, was equally or more efficient than inhibition of formins. Since actin polymerization and myosin contractility are linked in many cytoskeleton processes, additional careful analysis is needed in each case when function of formins was proposed solely on the basis of experiment with SMIFH2.

Impact of overweight and obesity on women with urinary incontinence: pilot study

IntroductionTo assess whether obesity has a greater impact than overweight on urinary incontinence severity, pelvic floor muscle function, and quality of life in women with urinary incontinence. MethodsA pilot cross-sectional study using a convenience sample. Twenty-six volunteers were evaluated and divided into: Overweight Group (n=11) with BMI (body mass index) between 25.0-29.9kg/m²; Obesity Group (n=15) BMI≥30.0kg/m². The volunteers performed the urogynecological evaluation, Incontinence Severity Index (ISI), the King’s Health Questionnaire (KHQ), 1-hour pad test and evaluation of pelvic floor muscle function. Statistical analysis: Shapiro–Wilk test and the Mann-Whitney test for intergroup analysis. The significance level: p≤0.05. ResultsThe average age was 61.09(12.51) in the Overweight Group and 55.93(9.03) years in the Obesity Group. The Overweight Group presented better fast fiber contraction (p=0.03) of the pelvic floor muscle. There were no differences in the ISI and quality of life between the groups. ConclusionsThere was no difference in the impact caused by being overweight or obese in relation to urinary incontinence severity, pelvic floor muscle function and quality of life, except for fast fiber contraction in which Overweight Group showed better results.

The Mechanical Contribution of Vimentin to Cellular Stress Generation

Contractile stress generation by adherent cells is largely determined by the interplay of forces within their cytoskeleton. It is known that actin stress fibers, connected to focal adhesions, provide contractile stress generation, while microtubules and intermediate filaments provide cells compressive stiffness. Recent studies have shown the importance of the interplay between the stress fibers and the intermediate filament vimentin. Therefore, the effect of the interplay between the stress fibers and vimentin on stress generation was quantified in this study. We hypothesized that net stress generation comprises the stress fiber contraction combined with the vimentin resistance. We expected an increased net stress in vimentin knockout (VimKO) mouse embryonic fibroblasts (MEFs) compared to their wild-type (vimentin wild-type (VimWT)) counterparts, due to the decreased resistance against stress fiber contractility. To test this, the net stress generation by VimKO and VimWT MEFs was determined using the thin film method combined with sample-specific finite element modeling. Additionally, focal adhesion and stress fiber organization were examined via immunofluorescent staining. Net stress generation of VimKO MEFs was three-fold higher compared to VimWT MEFs. No differences in focal adhesion size or stress fiber organization and orientation were found between the two cell types. This suggests that the increased net stress generation in VimKO MEFs was caused by the absence of the resistance that vimentin provides against stress fiber contraction. Taken together, these data suggest that vimentin resists the stress fiber contractility, as hypothesized, thus indicating the importance of vimentin in regulating cellular stress generation by adherent cells.

Dissipation of contractile forces: the missing piece in cell mechanics

Mechanical forces are key regulators of cell and tissue physiology. The basic molecular mechanism of fiber contraction by the sliding of actin filament upon myosin leading to conformational change has been known for decades. The regulation of force generation at the level of the cell, however, is still far from elucidated. Indeed, the magnitude of cell traction forces on the underlying extracellular matrix in culture is almost impossible to predict or experimentally control. The considerable variability in measurements of cell-traction forces indicates that they may not be the optimal readout to properly characterize cell contractile state and that a significant part of the contractile energy is not transferred to cell anchorage but instead is involved in actin network dynamics. Here we discuss the experimental, numerical, and biological parameters that may be responsible for the variability in traction force production. We argue that limiting these sources of variability and investigating the dissipation of mechanical work that occurs with structural rearrangements and the disengagement of force transmission is key for further understanding of cell mechanics.

IN-SILICO MODELING OF LEFT VENTRICLE TO SIMULATE DILATED CARDIOMYOPATHY AND CF-LVAD SUPPORT

Numerical modeling of the left ventricle dynamics plays an important role in testing different physiological scenarios and treatment techniques before the in vitro and in vivo assessments. However, utilized left ventricle model becomes vital in the simulations because validity of the results depends on the response of the numerical model to the parameter changes and additional sub-models for the applied treatment techniques. In this study, it is aimed to evaluate different numerical left ventricle models describing healthy and failing ventricle dynamics as well as the response of these models under continuous flow left ventricular assist device support. Six different numerical left ventricle models which include time varying elastance and single fiber contraction approaches are selected and applied in combination with a closed loop electric analogue of the circulation to achieve this purpose. The time varying elastace models relate ventricular pressure and volume changes in a simplistic way while the single fiber contraction models combine different scales ranging from protein to organ level. Change of the hemodynamic signals at the organ level for healthy, failing and CF-LVAD supported left ventricle models shows functionality of these models and helps to understand usability of them for different purposes.