Srf-dependent paracrine signals produced by myofibers control satellite cell-mediated skeletal muscle hypertrophy

Adult skeletal muscles adapt their fiber size to workload. We show that serum response factor (Srf) is required for satellite cell-mediated hypertrophic muscle growth. Deletion of Srf from myofibers and not satellite cells blunts overload-induced hypertrophy, and impairs satellite cell proliferation and recruitment to pre-existing fibers. We reveal a gene network in which Srf within myofibers modulates interleukin-6 and cyclooxygenase-2/interleukin-4 expressions and therefore exerts a paracrine control of satellite cell functions. In Srf-deleted muscles, in vivo overexpression of interleukin-6 is sufficient to restore satellite cell proliferation but not satellite cell fusion and overall growth. In contrast cyclooxygenase-2/interleukin-4 overexpression rescue satellite cell recruitment and muscle growth without affecting satellite cell proliferation, identifying altered fusion as the limiting cellular event. These findings unravel a role for Srf in the translation of mechanical cues applied to myofibers into paracrine signals, which in turn will modulate satellite cell functions and support muscle growth.     

Impact of active transport and transpiration on nickel and cadmium accumulation in the leaves of the Ni-hyperaccumulator Leptoplax emarginata: a biophysical approach

Aims

The primary aim of this study was to investigate the impact of active nickel and cadmium transport, transpiration and shoot biomass production on Ni and Cd accumulation in the leaves of the Ni-hyperaccumulator Leptoplax emarginata. A secondary objective was to observe the effects of various concentrations of nickel and cadmium in solutions on the plant growth and ecophysiological characteristics of these plants. Finally, the study sought to identify possible nickel and cadmium concentration gradients in solution as a function of the root distance.

Methods

The Intact Plant Transpiration Stream Concentration Factor (TSCF=xylem/solution solute concentration ratio) was determined for both Ni and Cd and for the selected intact transpiring Ni-hyperaccumulator Leptoplax emarginata, cultivated on two contrasting fertilized and Ni-Cd-contaminated sandy porous media (rhizotrons with central root compartments, linked to Mariotte tubes operated at ?1?kPa). IPTSCF_Ni and IPTSCF_Cd were calculated as the ratios between the hyperaccumulator plant’s nickel or cadmium mass in the leaves and the nickel or cadmium concentration in solution by the volume of water transpired during the period of culture. Plant growth characteristics and gas exchanges were also recorded.

Results

IPTSCF values were much greater than 1 (IPTSCF_Ni?=?5.2?±?0.9 and IPTSCF_Cd?=?4.4?±?0.6) whatever the amount of available Ni and Cd. This characterized a predominantly active plant metal uptake. Moreover, biological regulation was reported: plant growth and transpiration were significantly lower for hyperaccumulator plants cultivated in sand which was rich in available Ni and Cd, than for hyperaccumulator plants cultivated in topsoil, poor in available Ni and Cd. In the soil rhizosphere, capillary flow was related to transpiration and a depletion pattern was developed for Ni and sometimes for Cd.

Conclusions

Overall, the Intact Plant Transpiration Stream Concentration Factor appeared to be a relevant metal bioconcentration factor taking into account the predominant type of metal transport from roots to leaves, plant growth and transpiration coupling and metal availability. IPTSCF_Ni and IPTSCF_Cd values were much greater than 1 and similar whatever the amount of available Ni and Cd. This characterized a predominantly active plant combining Ni and Cd uptake and biological regulations dependent of the Ni and Cd concentrations in solution.     

Stress-induced phosphorylation and proteasomal degradation of mitofusin 2 facilitates mitochondrial fragmentation and apoptosis

Mitochondria play central roles in integrating pro- and antiapoptotic stimuli, and JNK is well known to have roles in activating apoptotic pathways. We establish a critical link between stress-induced JNK activation, mitofusin 2, which is an essential component of the mitochondrial outer membrane fusion apparatus, and the ubiquitin-proteasome system (UPS). JNK phosphorylation of mitofusin 2 in response to cellular stress leads to recruitment of the ubiquitin ligase (E3) Huwe1/Mule/ARF-BP1/HectH9/E3Histone/Lasu1 to mitofusin 2, with the BH3 domain of Huwe1 implicated in this interaction. This results in ubiquitin-mediated proteasomal degradation of mitofusin 2, leading to mitochondrial fragmentation and enhanced apoptotic cell death. The stability of a nonphosphorylatable mitofusin 2 mutant is unaffected by stress and protective against apoptosis. Conversely, a mitofusin 2 phosphomimic is more rapidly degraded without cellular stress. These findings demonstrate how proximal signaling events can influence both mitochondrial dynamics and apoptosis through phosphorylation-stimulated degradation of the mitochondrial fusion machinery.     

Protein kinase D1 is essential for contraction-induced glucose uptake but is not involved in fatty acid uptake into cardiomyocytes

Increased contraction enhances substrate uptake into cardiomyocytes via translocation of the glucose transporter GLUT4 and the long chain fatty acid (LCFA) transporter CD36 from intracellular stores to the sarcolemma. Additionally, contraction activates the signaling enzymes AMP-activated protein kinase (AMPK) and protein kinase D1 (PKD1). Although AMPK has been implicated in contraction-induced GLUT4 and CD36 translocation in cardiomyocytes, the precise role of PKD1 in these processes is not known. To study this, we triggered contractions in cardiomyocytes by electric field stimulation (EFS). First, the role of PKD1 in GLUT4 and CD36 translocation was defined. In PKD1 siRNA-treated cardiomyocytes as well as cardiomyocytes from PKD1 knock-out mice, EFS-induced translocation of GLUT4, but not CD36, was abolished. In AMPK siRNA-treated cardiomyocytes and cardiomyocytes from AMPKα2 knock-out mice, both GLUT4 and CD36 translocation were abrogated. Hence, unlike AMPK, PKD1 is selectively involved in glucose uptake. Second, we analyzed upstream factors in PKD1 activation. Cardiomyocyte contractions enhanced reactive oxygen species (ROS) production. Using ROS scavengers, we found that PKD1 signaling and glucose uptake are more sensitive to changes in intracellular ROS than AMPK signaling or LCFA uptake. Furthermore, silencing of death-activated protein kinase (DAPK) abrogated EFS-induced GLUT4 but not CD36 translocation. Finally, possible links between PKD1 and AMPK signaling were investigated. PKD1 silencing did not affect AMPK activation. Reciprocally, AMPK silencing did not alter PKD1 activation. In conclusion, we present a novel contraction-induced ROS-DAPK-PKD1 pathway in cardiomyocytes. This pathway is activated separately from AMPK and mediates GLUT4 translocation/glucose uptake, but not CD36 translocation/LCFA uptake.     

The Siva protein is a novel intracellular ligand of the CD4 receptor that promotes HIV-1 envelope-induced apoptosis in T-lymphoid cells

In addition to its positive signaling function in the antigen presentation process, CD4 acts as the primary receptor for HIV-1. Contact between CD4 and the viral envelope leads to virus entry, but can also trigger apoptosis of uninfected CD4⁺ T-cells through a mechanism that is poorly understood. We show that Siva-1, a death domain-containing proapoptotic protein, associates with the cytoplasmic domain of CD4. This interaction is mediated by the cysteine-rich region found in the C-terminal part of the Siva-1 protein. Expression of Siva-1 specifically increases the susceptibility of both T-cell lines and unstimulated human primary CD4⁺ T-lymphocytes to CD4-mediated apoptosis triggered by the HIV-1 envelope, and results in activation of a caspase-dependent mitochondrial pathway. The same susceptibility is observed in T-cells expressing a truncated form of CD4 that is able to recruit Siva-1 but fails to associate with p56^Lck, indicating that Siva-1 participates in a pathway independent of the p56^Lck kinase activity. Altogether, these results suggest that Siva-1 might participate in the CD4-initiated signaling apoptotic pathway induced by the HIV-1 envelope in T-lymphoid cells. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Chromatographic study of magnesium and calcium binding to immobilized human serum albumin

The use of immobilized human serum albumin (HSA) as a stationary phase in affinity chromatography has been shown to be useful in resolving optical antipodes or to investigate interactions between drugs and protein. However, to our knowledge, no inorganic ion binding has been studied on this immobilized protein type. To do this, the human serum albumin stationary phase was assimilated to a weak cation-exchanger by working with a mobile phase pH equal to 6.5. A study of the eluent ionic strength effect on ion retention was carried out by varying the buffer concentrations and the column temperatures. The thermodynamic parameters for magnesium and calcium transfer from the mobile to the stationary phase were determined from linear van’t Hoff plots. An enthalpy–entropy compensation study revealed that the type of interaction was independent of the mobile phase composition. A simple model based on the Gouy–Chapman theory was considered in order to describe the retention behavior of the test cations with the mobile phase ionic strength. From this theoretical approach, the relative charge densities of the human serum albumin surface implied in the binding process were estimated at different column temperatures.     

Mixing Languages during Learning? Testing the One Subject—One Language Rule

In bilingual communities, mixing languages is avoided in formal schooling: even if two languages are used on a daily basis for teaching, only one language is used to teach each given academic subject. This tenet known as the one subject-one language rule avoids mixing languages in formal schooling because it may hinder learning. The aim of this study was to test the scientific ground of this assumption by investigating the consequences of acquiring new concepts using a method in which two languages are mixed as compared to a purely monolingual method. Native balanced bilingual speakers of Basque and Spanish—adults (Experiment 1) and children (Experiment 2)—learnt new concepts by associating two different features to novel objects. Half of the participants completed the learning process in a multilingual context (one feature was described in Basque and the other one in Spanish); while the other half completed the learning phase in a purely monolingual context (both features were described in Spanish). Different measures of learning were taken, as well as direct and indirect indicators of concept consolidation. We found no evidence in favor of the non-mixing method when comparing the results of two groups in either experiment, and thus failed to give scientific support for the educational premise of the one subject—one language rule.     

Genetics of frost hardiness in <Emphasis Type="Italic">Juglans regia</Emphasis> L. and relationship with growth and phenology

The growing interest in broadleaf timber plantations in the Mediterranean area has promoted several studies focusing on the identification and characterization of variability sources in main timber-producing species. J. regia is one of these species, well-adapted to this area, but with freezing, damages registrations. Breeding focused on productive traits should include knowledge of adaptation, required to obtain a good selection capable of producing a suitable turnover in timber plantations. In this study, the features evaluated were autumn and winter frost hardiness and some vegetative traits on 22 half-sib J. regia progenies. Budsticks were exposed to sub-zero temperatures in a controlled chamber and using measurements of relative electrolyte content, the LT50 values (°C) were calculated by each individual. The study was carried out on seven-year-old progenies. The familiar heritability of autumn frost hardiness was 0.68, and on winter, it was 0.77. The autumn frost behaviour correlated genetically with the length of the growing season (0.574 ± 0.351), and both autumn and winter frost hardiness correlated inversely with secondary annual growth measured at breast height (?0.654 ± 0.259 and ?0.740 ± 0.227, respectively). These results pointed that growth could therefore be improved without increasing the frost vulnerability. This should be important for growers, particularly under climate change conditions.     

Consistent probabilistic outputs for protein function prediction

In predicting hierarchical protein function annotations, such as terms in the Gene Ontology (GO), the simplest approach makes predictions for each term independently. However, this approach has the unfortunate consequence that the predictor may assign to a single protein a set of terms that are inconsistent with one another; for example, the predictor may assign a specific GO term to a given protein ('purine nucleotide binding') but not assign the parent term ('nucleotide binding'). Such predictions are difficult to interpret. In this work, we focus on methods for calibrating and combining independent predictions to obtain a set of probabilistic predictions that are consistent with the topology of the ontology. We call this procedure 'reconciliation'. We begin with a baseline method for predicting GO terms from a collection of data types using an ensemble of discriminative classifiers. We apply the method to a previously described benchmark data set, and we demonstrate that the resulting predictions are frequently inconsistent with the topology of the GO. We then consider 11 distinct reconciliation methods: three heuristic methods; four variants of a Bayesian network; an extension of logistic regression to the structured case; and three novel projection methods - isotonic regression and two variants of a Kullback-Leibler projection method. We evaluate each method in three different modes - per term, per protein and joint - corresponding to three types of prediction tasks. Although the principal goal of reconciliation is interpretability, it is important to assess whether interpretability comes at a cost in terms of precision and recall. Indeed, we find that many apparently reasonable reconciliation methods yield reconciled probabilities with significantly lower precision than the original, unreconciled estimates. On the other hand, we find that isotonic regression usually performs better than the underlying, unreconciled method, and almost never performs worse; isotonic regression appears to be able to use the constraints from the GO network to its advantage. An exception to this rule is the high precision regime for joint evaluation, where Kullback-Leibler projection yields the best performance.