Kv3.1 uses a timely resurgent K+ current to secure action potential repolarization

High-frequency action potential (AP) transmission is essential for rapid information processing in the central nervous system. Voltage-dependent K(v)3 channels play an important role in this process thanks to their high activation threshold and fast closure kinetics, which reduce the neuron's refractory period. However, premature Kv3 channel closure leads to incomplete membrane repolarization, preventing sustainable AP propagation. Here, we demonstrate that K(v)3.1b channels solve this problem by producing resurgent K+ currents during repolarization, thus ensuring enough repolarizing power to terminate each AP. Unlike previously described resurgent Na+ and K+ currents, K(v)3.1b's resurgent current does not originate from recovery of channel block or inactivation but results from a unique combination of steep voltage-dependent gating kinetics and ultra-fast voltage-sensor relaxation. These distinct properties are readily transferrable onto an orthologue K-v channel by transplanting the voltage-sensor's S3-S4 loop, providing molecular insights into the mechanism by which K(v)3 channels contribute to high-frequency AP transmission

On the Role of the Difference in Surface Tensions Involved in the Allosteric Regulation of NHE-1 Induced by Low to Mild Osmotic Pressure, Membrane Tension and Lipid Asymmetry

The sodium-proton exchanger 1 (NHE-1) is a membrane transporter that exchanges Na+ for H+ ion across the membrane of eukaryotic cells. It is cooperatively activated by intracellular protons, and this allosteric regulation is modulated by the biophysical properties of the plasma membrane and related lipid environment. Consequently, NHE-1 is a mechanosensitive transporter that responds to osmotic pressure, and changes in membrane composition. The purpose of this study was to develop the relationship between membrane surface tension, and the allosteric balance of a mechanosensitive transporter such as NHE-1. In eukaryotes, the asymmetric composition of membrane leaflets results in a difference in surface tensions that is involved in the creation of a reservoir of intracellular vesicles and membrane buds contributing to buffer mechanical constraints. Therefore, we took this phenomenon into account in this study and developed a set of relations between the mean surface tension, membrane asymmetry, fluid phase endocytosis and the allosteric equilibrium constant of the transporter. We then used the experimental data published on the effects of osmotic pressure and membrane modification on the NHE-1 allosteric constant to fit these equations. We show here that NHE-1 mechanosensitivity is more based on its high sensitivity towards the asymmetry between the bilayer leaflets compared to mean global membrane tension. This compliance to membrane asymmetry is physiologically relevant as with their slower transport rates than ion channels, transporters cannot respond as high pressure-high conductance fast-gating emergency valves

The Effect of Sustained Compression on Oxygen Metabolic Transport in the Intervertebral Disc Decreases with Degenerative Changes

Intervertebral disc metabolic transport is essential to the functional spine and provides the cells with the nutrients necessary to tissue maintenance. Disc degenerative changes alter the tissue mechanics, but interactions between mechanical loading and disc transport are still an open issue. A poromechanical finite element model of the human disc was coupled with oxygen and lactate transport models. Deformations and fluid flow were linked to transport predictions by including strain-dependent diffusion and advection. The two solute transport models were also coupled to account for cell metabolism. With this approach, the relevance of metabolic and mechano-transport couplings were assessed in the healthy disc under loading-recovery daily compression. Disc height, cell density and material degenerative changes were parametrically simulated to study their influence on the calculated solute concentrations. The effects of load frequency and amplitude were also studied in the healthy disc by considering short periods of cyclic compression. Results indicate that external loads influence the oxygen and lactate regional distributions within the disc when large volume changes modify diffusion distances and diffusivities, especially when healthy disc properties are simulated. Advection was negligible under both sustained and cyclic compression. Simulating degeneration, mechanical changes inhibited the mechanical effect on transport while disc height, fluid content, nucleus pressure and overall cell density reductions affected significantly transport predictions. For the healthy disc, nutrient concentration patterns depended mostly on the time of sustained compression and recovery. The relevant effect of cell density on the metabolic transport indicates the disturbance of cell number as a possible onset for disc degeneration via alteration of the metabolic balance. Results also suggest that healthy disc properties have a positive effect of loading on metabolic transport. Such relation, relevant to the maintenance of the tissue functional composition, would therefore link disc function with disc nutrition

Influence des défauts cristallins sur les changements de phase induits par faisceaux d'ions dans les films minces d'oxyde d' yttrium Y2O3

Dans ce travail, nous avons étudié le comportement structural des films minces d'oxyde d'yttrium sous faisceau d'ions, lors de leur élaboration et sous irradiation. Une attention particulière a été portée sur le rôle des défauts cristallins en combinant les techniques de DRX, MET et RBS. Les films déposés sur différents substrats par pulvérisation ionique présentent un écart important par rapport à la structure cubique-C idéale, avec de nombreux défauts induits lors de la croissance par effet d' atomic peening (lacunes d'oxygène et paires anti-Frenkel). L'état de contrainte résultant a été décrit par un modèle triaxial, permettant d'interpréter les évolutions expérimentales observées. Les cinétiques de remise en ordre sous recuit de la structure défective ont été étudiées par DRX in situ. Une forte influence de la contrainte initiale (et donc de l'énergie élastique emmagasinée) a été clairement mise en évidence. Pour les forts états de contraintes résiduelles, la transformation est observée à basse température (dès 250C). Une approche thermodynamique justifie que la structure cubique-C défective peut être assimilée à une phase de type fluorine stabilisée par la contrainte. Enfin, deux comportements sont observés sous irradiation aux ions xénon, avec à basse énergie (<180 keV) une amorphisation de la structure, et à haute énergie une transition cubique-monoclinique. Pour justifier l'existence de ces deux régimes, le mécanisme de transformation a été expliqué par un effet de stabilisation, par la contrainte, de boucles lacunaires dans les plans {111}, de rayon sur-critique dépendant de la taille des cascades.In this work, we have investigated the structural behaviour of yttrium oxide thin films under ion beam, during the deposition process and under irradiation. A particular attention has been paid on the influence of crystal defects by combining XRD, TEM and RBS techniques. The films deposited using the ion beam sputtering deposition technique on different substrates show an important deviation in comparison with the ideal cubic-C structure, with the existence of many defects induced by the atomic peening effect during the growth (oxygen vacancies and anti-Frenkel pairs). The resulting state of stress has been described using a triaxial model which allows to explain experimental evolutions. The reordering kinetics of the defective structure under annealing have been studied by mean of in situ XRD. A strong influence of the initial stress (and so of the stored elastic energy) has clearly been shown. For the higher residual stress states, the transformation is observed at low temperature (from 250C). A thermodynamic approach justify that the defective cubic-C structure could be seen as a fluorite-like phase stabilized by the stress. Finally, two behaviours have been observed under xenon ion irradiation with, at low energy (<180keV) an amorphization of the structure, and at high energy a cubic-to-monoclinic transition. To explain these two regimes, the transformation mechanism has been linked to the stabilization, by the stress, of vacancy loops in the {111} planes, with supercritical radius depending on the cascade size.POITIERS-BU Sciences (861942102) / SudocSudocFranceF

Force-induced motions of the PIEZO1 blade probed with fluorimetry

Summary: Mechanical forces are thought to activate mechanosensitive PIEZO channels by changing the conformation of a large transmembrane blade domain. Yet, whether different stimuli induce identical conformational changes in this domain remains unclear. Here, we repurpose a cyclic permuted green fluorescent protein as a conformation-sensitive probe to track local rearrangements along the PIEZO1 blade. Two independent probes, one inserted in an extracellular site distal to the pore and the other in a distant intracellular proximal position, elicit sizable fluorescence signals when the tagged channels activate in response to fluid shear stress of low intensity. Neither cellular indentations nor osmotic swelling of the cell elicit detectable fluorescence signals from either probe, despite the ability of these stimuli to activate the tagged channels. High-intensity flow stimuli are ineffective at eliciting fluorescence signals from either probe. Together, these findings suggest that low-intensity fluid shear stress causes a distinct form of mechanical stress to the cell

S3-S4 linker length modulates the relaxed state of a voltage-gated potassium channel

Voltage-sensing domains (VSDs) are membrane protein modules found in ion channels and enzymes that are responsible for a large number of fundamental biological tasks, such as neuronal electrical activity. The VSDs switch from a resting to an active conformation upon membrane depolarization, altering the activity of the protein in response to voltage changes. Interestingly, numerous studies describe the existence of a third distinct state, called the relaxed state, also populated at positive potentials. Although some physiological roles for the relaxed state have been suggested, little is known about the molecular determinants responsible for the development and modulation of VSD relaxation. Several lines of evidence have suggested that the linker (S3-S4 linker) between the third (S3) and fourth (S4) transmembrane segments of the VSD alters the equilibrium between resting and active conformations. By measuring gating currents from the Shaker potassium channel, we demonstrate here that shortening the S3-S4 linker stabilizes the relaxed state, whereas lengthening the linker or splitting it and coinjecting two fragments of the channel have little effect. We propose that natural variations of the length of the S3-S4 linker in various VSD-containing proteins may produce differential VSD relaxation in vivo

A CeO2/PVDC hybrid latex mediated by a phosphonated macro-RAFT agent

A poly(vinylidene chloride-co-Me acrylate) hybrid latex comprising CeO2 nanoparticles was successfully prepd. by emulsion polymn. employing a water-sol. phosphonated macro-RAFT agent. A poly(vinylbenzylphosphonic diacid-co-styrene) statistical copolymer was first synthesized, using dibenzyl trithiocarbonate as a controlling agent, and adsorbed on ceria nanoparticles. UV-visible and 31P NMR spectroscopy proved to be efficient and complementary techniques to assess the extent of interactions between the copolymer and ceria nanoparticles, leading to a better understanding of the adsorption of phosphonated copolymer chains on inorg. particles. Then, these functionalized-CeO2 nanoparticles were used to mediate the emulsion copolymn. of vinylidene chloride and Me acrylate in the presence of a very low amt. of emulsifier. Cryo-transmission electron microscopy (cryo-TEM) confirmed the hybrid structure of the latex and the absence of either free ceria nanoparticles or free PVDC latex particles