Activity of TREK-2-like Channels in the Pyramidal Neurons of Rat Medial Prefrontal Cortex Depends on Cytoplasmic Calcium

TREK-2-like channels in the pyramidal neurons of rat prefrontal cortex are characterized by a wide range of spontaneous activity—from very low to very high—independent of the membrane potential and the stimuli that are known to activate TREK-2 channels, such as temperature or membrane stretching. The aim of this study was to discover what factors are involved in high levels of TREK-2-like channel activity in these cells. Our research focused on the PI(4,5)P2-dependent mechanism of channel activity. Single-channel patch clamp recordings were performed on freshly dissociated pyramidal neurons of rat prefrontal cortexes in both the cell-attached and inside-out configurations. To evaluate the role of endogenous stimulants, the activity of the channels was recorded in the presence of a PI(4,5)P2 analogue (PI(4,5)P2DiC8) and Ca2+. Our research revealed that calcium ions are an important factor affecting TREK-2-like channel activity and kinetics. The observation that calcium participates in the activation of TREK-2-like channels is a new finding. We showed that PI(4,5)P2-dependent TREK-2 activity occurs when the conditions for PI(4,5)P2/Ca2+ nanocluster formation are met. We present a possible model explaining the mechanism of calcium action.

Cellular transduction of mechanical oscillations in plants by the plasma-membrane mechanosensitive channel MSL10

Plants spend most of their life oscillating around 1–3 Hz due to the effect of the wind. Therefore, stems and foliage experience repetitive mechanical stresses through these passive movements. However, the mechanism of the cellular perception and transduction of such recurring mechanical signals remains an open question. Multimeric protein complexes forming mechanosensitive (MS) channels embedded in the membrane provide an efficient system to rapidly convert mechanical tension into an electrical signal. So far, studies have mostly focused on nonoscillatory stretching of these channels. Here, we show that the plasma-membrane MS channel MscS-LIKE 10 (MSL10) from the model plant Arabidopsis thaliana responds to pulsed membrane stretching with rapid activation and relaxation kinetics in the range of 1 s. Under sinusoidal membrane stretching MSL10 presents a greater activity than under static stimulation. We observed this amplification mostly in the range of 0.3–3 Hz. Above these frequencies the channel activity is very close to that under static conditions. With a localization in aerial organs naturally submitted to wind-driven oscillations, our results suggest that the MS channel MSL10, and by extension MS channels sharing similar properties, represents a molecular component allowing the perception of oscillatory mechanical stimulations by plants.

Transduction of mechanical cellular oscillation by the plasma-membrane mechanosensitive channel MSL10

Throughout their life, plants are submitted to recurrent cyclic mechanical loading due to wind. The resulting passive oscillation movements of stem and foliage is an important phenomenon for biological and ecological issues such as photosynthesis optimization 1–3 and thermal exchange 4. The induced motions at plant scale are well described and analyzed, with oscillations at typically 1 to 3 Hz in trees 5–10. However, the cellular perception and transduction of such recurring mechanical signals remains an open question. Multimeric protein complexes forming mechanosensitive (MS) channels embedded in the membrane provide an efficient system to rapidly convert mechanical tension into electrical signal 11. Here we show that the plasma membrane mechanosensitive channel MscS-LIKE 10 (MSL10) from the model plant Arabidopsis thaliana responds to pulsed membrane stretching with rapid activation and relaxation kinetics in the range of one second. Under sinusoidal membrane stretching MSL10 presents a greater activity than under static stimulation and behaves as a large bandpass oscillation “follower” without filtering the signal in the range of 0.3 to 3 Hz. With a localization in aerial organs naturally submitted to oscillations, our results suggest that the mechanosensitive channel MSL10 represents a molecular component of a universal system of oscillatory perception in plants.

Temperature-sensitive mutants of MscL mechanosensitive channel

MscL is a mechanosensitive channel that undergoes a global conformational change upon application of membrane stretching. To elucidate how the structural stability and flexibility occur, we isolated temperature-sensitive (Ts) mutants of Escherichia coli MscL that allowed cell growth at 32°C but not at 42°C. Two Ts mutants, L86P and D127V, were identified. The L86P mutation occurred in the second transmembrane helix, TM2. Substitution of residues neighbouring L86 with proline also led to a Ts mutation, but the substitution of L86 with other amino acids did not result in a Ts phenotype, indicating that the Ts phenotype was due to a structural change of TM2 helix by the introduction of a proline residue. The D127V mutation was localized in the electrostatic belt of the bundle of cytoplasmic helices, indicating that stability of the pentameric bundle of the cytoplasmic helix affects MscL structure. Together, this study described a novel class of MscL mutations that were correlated with the thermodynamic stability of the MscL structure.

On the Role of Fluid–Structure Interaction on Structural Loading by Pressure Waves in Air

This study investigates the significance of fluid–structure interaction (FSI) effects on structural response to pressure wave and shock wave loading. Finite element (FE) simulations and one-dimensional (1D) analytical models are used to compare the responses of simple structures in presence and absence of FSI. Results are provided in nondimensional form and allow rapid estimation of the significance of FSI. The cases of a square elastic plate in bending and a square rigid-perfectly plastic plate undergoing membrane stretching are discussed in detail. We deduce simple formulae to identify scenarios in which effects of FSI can be neglected.

Propagation of a viscous thin film over an elastic membrane

We study the buoyancy-driven spreading of a thin viscous film over a thin elastic membrane. Neglecting the effects of membrane bending and the membrane weight, we study the case of constant fluid injection and obtain a system of coupled partial differential equations to describe the shape of the air–liquid interface, and the deformation and radial tension of the stretched membrane. We obtain self-similar solutions to describe the dynamics. In particular, in the early-time period, the dynamics is dominated by buoyancy-driven spreading of the liquid film, and membrane stretching is a response to the buoyancy-controlled distribution of liquid weight; the location of the liquid front obeys the power-law form $r_{f}(t)\propto t^{1/2}$. However, in the late-time period, the system is quasi-steady, the air–liquid interface is flat, and membrane stretching, due to the liquid weight, causes the spreading of the liquid front; the location of the front obeys a different power-law form $r_{f}(t)\propto t^{1/4}$ before the edge effects of the membrane become significant. In addition, we report laboratory experiments for constant fluid injection using different viscous liquids and thin elastic membranes. Very good agreement is obtained between the theoretical predictions and experimental observations.

A phosphoinositide-binding cluster in cavin1 acts as a molecular sensor for cavin1 degradation

Caveolae are abundant surface organelles implicated in a range of cellular processes. Two classes of proteins work together to generate caveolae: integral membrane proteins termed caveolins and cytoplasmic coat proteins called cavins. Caveolae respond to membrane stress by releasing cavins into the cytosol. A crucial aspect of this model is tight regulation of cytosolic pools of cavin under resting conditions. We now show that a recently identified region of cavin1 that can bind phosphoinositide (PI) lipids is also a major site of ubiquitylation. Ubiquitylation of lysines within this site leads to rapid proteasomal degradation. In cells that lack caveolins and caveolae, cavin1 is cytosolic and rapidly degraded as compared with cells in which cavin1 is associated with caveolae. Membrane stretching causes caveolar disassembly, release of cavin complexes into the cytosol, and increased proteasomal degradation of wild-type cavin1 but not mutant cavin1 lacking the major ubiquitylation site. Release of cavin1 from caveolae thus leads to exposure of key lysine residues in the PI-binding region, acting as a trigger for cavin1 ubiquitylation and down-regulation. This mutually exclusive PI-binding/ubiquitylation mechanism may help maintain low levels of cytosolic cavin1 in resting cells, a prerequisite for cavins acting as signaling modules following release from caveolae.

An elastocapillary model of wood-fibre collapse

An elastocapillary model for drying-induced collapse is proposed. We consider a circular elastic membrane with a hole at the centre that is deformed by the capillary pressure of simply and doubly connected menisci. The membrane overlays a cylindrical cavity with rigid walls, trapping a prescribed volume of water. This geometry may be suitable for studying structural failures and stiction in micro-electromechanical systems during wet etching, where capillary surfaces experience catastrophic transitions. The dry state is determined using the dihedral-angle and volume-turning-point stability criteria. Open and collapsed conformations are predicted from the scaled hole radius, cavity aspect ratio, meniscus contact angle with the membrane and cavity walls, and an elastocapillary number measuring the membrane stretching rigidity relative to the water surface tension. For a given scaled hole radius and cavity aspect ratio, there is a critical elastocapillary number above which the system does not collapse upon drying. The critical elastocapillary number is weakly influenced by the contact angle over a wide range of the scaled hole radius, thus indicating a limitation of surface hydrophobization for controlling the dry-state conformation. The model is applied to the drying of wood fibres above the fibre saturation point, determining the conditions leading to collapse.