Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis.

Compensatory endocytosis keeps the membrane surface area of secretory cells constant following exocytosis. At chemical synapses, clathrin-independent ultrafast endocytosis maintains such homeostasis. This endocytic pathway is temporally and spatially coupled to exocytosis; it initiates within 50 ms at the region immediately next to the active zone where vesicles fuse. However, the coupling mechanism is unknown. Here, we demonstrate that filamentous actin is organized as a ring, surrounding the active zone at mouse hippocampal synapses. Assuming the membrane area conservation is due to this actin ring, our theoretical model suggests that flattening of fused vesicles exerts lateral compression in the plasma membrane, resulting in rapid formation of endocytic pits at the border between the active zone and the surrounding actin-enriched region. Consistent with model predictions, our data show that ultrafast endocytosis requires sufficient compression by exocytosis of multiple vesicles and does not initiate when actin organization is disrupted, either pharmacologically or by ablation of the actin-binding protein Epsin1. Our work suggests that membrane mechanics underlie the rapid coupling of exocytosis to endocytosis at synapses.

Lipid flip-flop and desorption from supported lipid bilayers is independent of curvature.

Flip-flop of lipids of the lipid bilayer (LBL) constituting the plasma membrane (PM) plays a crucial role in a myriad of events ranging from cellular signaling and regulation of cell shapes to cell homeostasis, membrane asymmetry, phagocytosis, and cell apoptosis. While extensive research has been conducted to probe the lipid flip flop of planar lipid bilayers (LBLs), less is known regarding lipid flip-flop for highly curved, nanoscopic LBL systems despite the vast importance of membrane curvature in defining the morphology of cells and organelles and in maintaining a variety of cellular functions, enabling trafficking, and recruiting and localizing shape-responsive proteins. In this paper, we conduct molecular dynamics (MD) simulations to study the energetics, structure, and configuration of a lipid molecule undergoing flip-flop and desorption in a highly curved LBL, represented as a nanoparticle-supported lipid bilayer (NPSLBL) system. We compare our findings against those of a planar substrate supported lipid bilayer (PSSLBL). Our MD simulation results reveal that despite the vast differences in the curvature and other curvature-dictated properties (e.g., lipid packing fraction, difference in the number of lipids between inner and outer leaflets, etc.) between the NPSLBL and the PSSLBL, the energetics of lipid flip-flop and lipid desorption as well as the configuration of the lipid molecule undergoing lipid flip-flop are very similar for the NPSLBL and the PSSLBL. In other words, our results establish that the curvature of the LBL plays an insignificant role in lipid flip-flop and desorption.

Formation and Properties of a Self-Assembled Nanoparticle-Supported Lipid Bilayer Probed through Molecular Dynamics Simulations.

We have carried out coarse-grained molecular dynamics (MD) simulations to study the self-assembly procedure of a system of randomly placed lipid molecules, water beads, and a nanoparticle (NP). The self-assembly results in the formation of the nanoparticle-supported lipid bilayer (NPSLBL), with the self-assembly mechanism being driven by events such as the formation of small lipid clusters, merging of the lipid clusters in the vicinity of the NP to form NP-embedded vesicle with a pore, and collapsing of that pore to eventually form the equilibrated NPSLBL system overcoming a large free-energy barrier. Subsequently, we quantify the properties and the configurations of this NPSLBL system. We reveal that unlike our proposition of an equal number of lipid molecules occupying the inner and outer leaflets in a recent report studying the properties of a preassembled lipid bilayer, the equilibrated self-assembled NPSLBL system demonstrates a much larger number of lipid molecules occupying the outer leaflet as compared to the inner leaflet. Second, the thickness of the water layer entrapped between the NP and the inner leaflet shows similar values as predicted by experiments and our previous study. Finally, we reveal that, similar to our previous study, the diffusivity of the lipid molecules in the outer leaflet is larger than that in the inner leaflet but, due to higher temperature employed during our simulations, are even larger than that predicted by our previous study.

Interactions of gold and silica nanoparticles with plasma membranes get distinguished by the van der Waals forces: Implications for drug delivery, imaging, and theranostics.

Making a nanoparticle (NP) approach and interact with a plasma membrane (PM) through the receptor-ligand interaction is key for applications like targeted drug delivery, cellular imaging, and theranostics. In this paper, we show that the van der Waals (vdW) interactions dominate the electrostatics ensuring that a gold NP approached the PM more spontaneously as compared to a silica NP. The negative σ (charge density) of a PM induces a negative electrostatic potential at the surface of the approaching gold NP and the silica NP; however, there is very little difference between these induced values due to a small electric double layer at the physiological salt concentration (c∞). Hence there is very little difference in the electrostatic repulsion between the two cases, while the PM-NP vdW attraction is much more for the gold NP as a result of a larger Hamaker constant. Therefore, in comparison to the gold NP, the silica NP would (a) undergo a promotion of the specific adhesion and a prevention of the non-specific adhesion simultaneously for a larger σ - c∞ phase space including the physiological conditions, (b) necessitate a larger length of the ligands to trigger spontaneous receptor-ligand interactions, and (c) require a larger driving force for force-driven receptor-ligand interactions.

Nanovesicles Versus Nanoparticle-Supported Lipid Bilayers: Massive Differences in Bilayer Structures and in Diffusivities of Lipid Molecules and Nanoconfined Water.

We carry out molecular dynamics (MD) simulations to compare the equilibrium architecture and properties of nanoparticle-supported lipid bilayers (NPSLBLs) with the free vesicles of similar dimensions. Three key differences emerge. First, we witness that for a free vesicle, a much larger number of lipid molecules occupy the outer layer as compared to the inner layer; on the other hand, for the NPSLBL the number of lipid molecules occupying the inner and outer layers is identical. Second, we witness that the diffusivities of the lipid molecules occupying both the inner and the outer layers of the free vesicles are identical, whereas for the NPSLBLs the diffusivity of the lipid molecules in the outer layer is more than twice the diffusivity of the lipid molecules in the inner layer. Finally, the NPSLBLs entrap nanoscopic thin water film between the inner lipid layer and the NP and the diffusivity of this water film is nearly 1 order of magnitude smaller than the diffusivity of the bulk water; on the other hand, the water inside the free vesicles has a diffusivity that is only slightly lower than that of the bulk water. Our findings, possibly the first probing the atomistic details of the NPSLBLs, are anticipated to shed light on the properties of this important nanomaterial with applications in a large number of disciplines ranging from drug and gene delivery to characterizing curvature-sensitive molecules.

Highly enhanced liquid flows via thermoosmotic effects in soft and charged nanochannels.

Enhancing nanoscale liquid flows remains an existing challenge in nanofluidics. Here we propose the generation of highly augmented thermoosmotic (TOS) liquid flows in soft nanochannels (or nanochannels functionalized by grafting with end-charged polyelectrolyte or PE brushes) by employing an axial temperature gradient. The TOS transport is a combination of the induced-electric-field electroosmotic (EOS) transport and a thermo-chemioosmotic (TCOS) transport with the latter resulting from an induced pressure gradient on account of the changes associated with the imposition of the axial temperature gradient. The end-charged brushes develop an electric double layer (EDL) localized at the charged, non-grafted brush end. Depending on the system parameters, this EDL localization massively augments the influence of the EOS body force and the induced pressure-gradient resulting in a TOS transport in soft nanochannels that can be more than one order of magnitude larger than that in brush-free nanochannels. Given the existing notion that the presence of the brushes invariably reduces the flow strength, this result of massive flow augmentation is extremely significant and non-trivial serving as a paradigm shift in the study of liquid transport in brush-grafted nanochannels.

Ion at Air-Water Interface Enhances Capillary Wave Fluctuations: Energetics of Ion Adsorption.

Recent simulations provide the energetics of ion adsorption at the air-water (a/w) interface: The presence of the ion at the interface suppresses the fluctuations of the capillary waves (CWs) reducing the entropy and displaces the weakly interacting water molecules to the bulk causing a reduction in the enthalpy. Here, we provide atomistic simulation-based evidence that the inferences of the existing studies stem from considering a small simulation volume that pins the CWs. For an appropriate size of the simulation system, an ion at the a/w interface enhances the CW fluctuations. Furthermore, we discover that the characteristics of the waves governing these enhanced CW fluctuations ensure a significant decrease in the pressure-volume work causing the enthalpy decrease, while the same wave characteristics of the CWs become responsible for an entropy decrease. Overall, the paper revisits the free energy picture of this fundamental problem of ion adsorption at the a/w interface.

Ionic Diffusoosmosis in Nanochannels Grafted with End-Charged Polyelectrolyte Brushes.

In this paper, we develop a theory to study the imposed axial salt-concentration-gradient-driven ionic diffusioosmosis (IDO) in soft nanochannels or nanochannels grafted with end-charged polyelectrolyte (PE) brushes. Our analysis first quantifies the diffusioosmotically induced electric field, which is primarily dictated by the imposed concentration gradient (CG) with little contribution of the induced osmosis. This induced electric field triggers an electroosmotic (EOS) transport, while the net diffusioosmotic (DOS) transport results from a combination of this EOS transport and a chemiosmotic (COS) transport arising from the pressure gradient induced by the applied CG. Our results demonstrate that the DOS transport is massively enhanced in nanochannels grafted with PE brushes with weak grafting density stemming from the significantly enhanced EOS transport caused by the localization of the EOS body force away from the nanochannel walls. This augmentation is even stronger for cases where the COS transport aids the EOS transport. On the other hand, the DOS transport gets severely reduced in nanochannels grafted with dense PE brushes owing to the severity of the brush-induced additional drag force. We anticipate that these findings will help to unravel an entirely new understanding of induced electrokinetic transport in soft nanochannels.

Theory of diffusioosmosis in a charged nanochannel.

We probe the diffusioosmotic transport in a charged nanofluidic channel in the presence of an applied tangential salt concentration gradient. Ionic salt gradient driven diffusioosmosis or ionic diffusioosmosis (IDO) is characterized by the generation of an induced tangential electric field and a diffusioosmotic velocity (DOSV) that is a combination of an electroosmotic velocity (EOSV) triggered by this electric field and a chemiosmotic velocity (COSV) triggered by an induced tangential pressure gradient. We explain that unlike the existing theories on IDO, it is more appropriate to apply the zero net current conditions (formulation F2) and not more restrictive zero net local flux conditions (formulation F1) particularly for the case where one considers a nanochannel connected to two reservoirs. We pinpoint limitations in the existing literature in correctly predicting the diffusioosmotic behavior even for the case where formulation F1 is used. We address these limitations and establish that (a) the induced electric field is an interplay of the differences in ionic diffusivity, the EDL-induced imbalance in ion concentrations, and the advection effects, (b) formulation F1 may overpredict or underpredict the electric field and the EOSV leading to an overprediction/underprediction of the DOSV and (c) formulation F2 demonstrates remarkable fluid physics of localized backflows owing to a dominant local influence of the COSV, which is missed by formulation F1. We anticipate that our theory will provide the first rigorous understanding of nanofluidic IDO with applications in multiple areas of low Reynolds number transport such as biofluidics, microfluidic separation, and colloidal transport.

Electric double layer electrostatics of lipid-bilayer-encapsulated nanoparticles: Toward a better understanding of protocell electrostatics.

Lipid-bilayer-encapsulated nanoparticles (LBLENPs) or NP-supported LBL systems, such as protocells (which are lipid bilayer encapsulated mesoporous silica nanoparticles or MSNPs) have received extensive attention for applications like targeted drug and gene deliveries, multimodal diagnostics, characterization of membrane-geometry sensitive molecules, etc. Very often electrostatic-mediated interactions have been hypothesized to play key roles in the functioning of these LBLENPs. Despite that, very little has been done to theoretically quantify the fundamental electric double layer (EDL) electrostatics of such LBLENPs. In this study, we develop an EDL theory to describe the electrostatics of such LBLENPs. We show that the electrostatics is a manifestation of the charged/dielectric nature of the NP, LBL structure and charging, and the ionic environment in which the LBLENPs are present. We also establish that for certain conditions of charging of the NP one witnesses a most remarkable charge inversion like electrostatics within the LBL membrane or the NP itself. We anticipate that our findings will provide an extremely useful platform for better understanding the fabrication and functioning of such LBLENPs and discuss examples where our theory can be useful.

Elasto-electro-capillarity: drop equilibrium on a charged, elastic solid.

We present a study here on elasto-electro-capillarity - for the first time, the matter of drop equilibrium on a soft (elastic and incompressible) and charged solid has been studied. Charges on the elastic solid induce an electric double layer or EDL at the solid-drop interface. Our analysis accounts for the electrostatic wetting contribution of the EDL in the overall energy balance. Our results reveal that (a) with an increase in "softness", the equilibrium solid-liquid contact angles show transition from the EDL-modified Young's law (rigid limit) to the EDL-modified Neumann's law (soft limit); (b) the EDL effects invariably enhance solid deformation and lower the apparent contact angle made by the drop with the undeformed solid; (c) the solid contact angles increase and the cusp made by the deformed solid undergoes enhanced rotation due to the EDL effects; and (d) the EDL effects are more prominent for the case where the solid-vapor surface energy exceeds the solid-liquid surface energy. The fact that the EDL effects invariably decrease the overall wetting energy of the system, thereby supporting a larger increase in the elastic energy associated with a larger solid deformation, explains all these findings and establishes that surface charges enhance the "softness" of a soft surface in the context of elastocapillarity.