Vesicle dynamics in elongation flow: Wrinkling instability and bud formation

We present experimental results on the relaxation dynamics of vesicles subjected to a time-dependent elongation flow. We observed and characterized a new instability, which results in the formation of higher order modes of the vesicle shape (wrinkles), after a switch in the direction of the gradient of the velocity. This surprising generation of membrane wrinkles can be explained by the appearance of a negative surface tension during the vesicle deflation, due to compression in a sign-switching transient. Moreover, the formation of buds in the vesicle membrane has been observed in the vicinity of the dynamical transition point.Comment: 4 pages, 4 figure

Effective Area-Elasticity and Tension of Micro-manipulated Membranes

We evaluate the effective Hamiltonian governing, at the optically resolved scale, the elastic properties of micro-manipulated membranes. We identify floppy, entropic-tense and stretched-tense regimes, representing different behaviors of the effective area-elasticity of the membrane. The corresponding effective tension depends on the microscopic parameters (total area, bending rigidity) and on the optically visible area, which is controlled by the imposed external constraints. We successfully compare our predictions with recent data on micropipette experiments.Comment: To be published in Phys. Rev. Let

The field theory of symmetrical layered electrolytic systems and the thermal Casimir effect

We present a general extension of a field-theoretic approach developed in earlier papers to the calculation of the free energy of symmetrically layered electrolytic systems which is based on the Sine-Gordon field theory for the Coulomb gas. The method is to construct the partition function in terms of the Feynman evolution kernel in the Euclidean time variable associated with the coordinate normal to the surfaces defining the layered structure. The theory is applicable to cylindrical systems and its development is motivated by the possibility that a static van der Waals or thermal Casimir force could provide an attractive force stabilising a dielectric tube formed from a lipid bilayer, an example of which are t-tubules occurring in certain muscle cells. In this context, we apply the theory to the calculation of the thermal Casimir effect for a dielectric tube of radius $R$ and thickness $\delta$ formed from such a membrane in water. In a grand canonical approach we find that the leading contribution to the Casimir energy behaves like $-k_BTL\kappa_C/R$ which gives rise to an attractive force which tends to contract the tube radius. We find that $\kappa_C \sim 0.3$ for the case of typical lipid membrane t-tubules. We conclude that except in the case of a very soft membrane this force is insufficient to stabilise such tubes against the bending stress which tend to increase the radius. We briefly discuss the role of lipid membrane reservoir implicit in the approach and whether its nature in biological systems may possibly lead to a stabilising mechanism for such lipid tubes.Comment: 28 pages, 2 figures, LaTe

Membrane shape as a reporter for applied forces

Recent advances have enabled 3-dimensional reconstructions of biological structures in vivo, ranging in size and complexity from single proteins to multicellular structures. In particular, tomography and confocal microscopy have been exploited to capture detailed 3-dimensional conformations of membranes in cellular processes ranging from viral budding and organelle maintenance to phagocytosis. Despite the wealth of membrane structures available, there is as yet no generic, quantitative method for their interpretation. We propose that by modeling these observed biomembrane shapes as fluid lipid bilayers in mechanical equilibrium, the externally applied forces as well as the pressure, tension, and spontaneous curvature can be computed directly from the shape alone. To illustrate the potential power of this technique, we apply an axial force with optical tweezers to vesicles and explicitly demonstrate that the applied force is equal to the force computed from the membrane conformation

A lattice model for the kinetics of rupture of fluid bilayer membranes

We have constructed a model for the kinetics of rupture of membranes under tension, applying physical principles relevant to lipid bilayers held together by hydrophobic interactions. The membrane is characterized by the bulk compressibility (for expansion), the thickness of the hydrophobic part of the bilayer, the hydrophobicity and a parameter characterizing the tail rigidity of the lipids. The model is a lattice model which incorporates strain relaxation, and considers the nucleation of pores at constant area, constant temperature, and constant particle number. The particle number is conserved by allowing multiple occupancy of the sites. An equilibrium ``phase diagram'' is constructed as a function of temperature and strain with the total pore surface and distribution as the order parameters. A first order rupture line is found with increasing tension, and a continuous increase in proto-pore concentration with rising temperature till instability. The model explains current results on saturated and unsaturated PC lipid bilayers and thicker artificial bilayers made of diblock copolymers. Pore size distributions are presented for various values of area expansion and temperature, and the fractal dimension of the pore edge is evaluated.Comment: 15 pages, 8 figure

Fluid-membrane tethers: minimal surfaces and elastic boundary layers

Thin cylindrical tethers are common lipid bilayer membrane structures, arising in situations ranging from micromanipulation experiments on artificial vesicles to the dynamic structure of the Golgi apparatus. We study the shape and formation of a tether in terms of the classical soap-film problem, which is applied to the case of a membrane disk under tension subject to a point force. A tether forms from the elastic boundary layer near the point of application of the force, for sufficiently large displacement. Analytic results for various aspects of the membrane shape are given.Comment: 12 page

Interactions between proteins bound to biomembranes

We study a physical model for the interaction between general inclusions bound to fluid membranes that possess finite tension, as well as the usual bending rigidity. We are motivated by an interest in proteins bound to cell membranes that apply forces to these membranes, due to either entropic or direct chemical interactions. We find an exact analytic solution for the repulsive interaction between two similar circularly symmetric inclusions. This repulsion extends over length scales of order tens of nanometers, and contrasts with the membrane-mediated contact attraction for similar inclusions on tensionless membranes. For non circularly symmetric inclusions we study the small, algebraically long-ranged, attractive contribution to the force that arises. We discuss the relevance of our results to biological phenomena, such as the budding of caveolae from cell membranes and the striations that are observed on their coats.Comment: 22 pages, 2 figure

Membranes by the Numbers

Many of the most important processes in cells take place on and across membranes. With the rise of an impressive array of powerful quantitative methods for characterizing these membranes, it is an opportune time to reflect on the structure and function of membranes from the point of view of biological numeracy. To that end, in this article, I review the quantitative parameters that characterize the mechanical, electrical and transport properties of membranes and carry out a number of corresponding order of magnitude estimates that help us understand the values of those parameters.Comment: 27 pages, 12 figure

Morphology and Interaction between Lipid Domains

Cellular membranes are a heterogeneous mix of lipids, proteins and small molecules. Special groupings of saturated lipids and cholesterol form a liquid-ordered phase, known as `lipid rafts,' serving as platforms for signaling, trafficking and material transport throughout the secretory pathway. Questions remain as to how the cell maintains heterogeneity of a fluid membrane with multiple phases, through time, on a length-scale consistent with the fact that no large-scale phase separation is observed. We have utilized a combination of mechanical modeling and in vitro experiments to show that membrane morphology can be a key player in maintaining this heterogeneity and organizing such domains in the membrane. We demonstrate that lipid domains can adopt a flat or dimpled morphology, where the latter facilitates a repulsive interaction that slows coalescence and tends to organize domains. These forces, that depend on domain morphology, play an important role in regulating lipid domain size and in the lateral organization of lipids in the membrane.Comment: 7 pages, 4 figure

Caspase-8 binding to cardiolipin in giant unilamellar vesicles provides a functional docking platform for bid

Caspase-8 is involved in death receptor-mediated apoptosis in type II cells, the proapoptotic programme of which is triggered by truncated Bid. Indeed, caspase-8 and Bid are the known intermediates of this signalling pathway. Cardiolipin has been shown to provide an anchor and an essential activating platform for caspase-8 at the mitochondrial membrane surface. Destabilisation of this platform alters receptor-mediated apoptosis in diseases such as Barth Syndrome, which is characterised by the presence of immature cardiolipin which does not allow caspase-8 binding. We used a simplified in vitro system that mimics contact sites and/or cardiolipin-enriched microdomains at the outer mitochondrial surface in which the platform consisting of caspase-8, Bid and cardiolipin was reconstituted in giant unilamellar vesicles. We analysed these vesicles by flow cytometry and confirm previous results that demonstrate the requirement for intact mature cardiolipin for caspase-8 activation and Bid binding and cleavage. We also used confocal microscopy to visualise the rupture of the vesicles and their revesiculation at smaller sizes due to alteration of the curvature following caspase-8 and Bid binding. Biophysical approaches, including Laurdan fluorescence and rupture/tension measurements, were used to determine the ability of these three components (cardiolipin, caspase-8 and Bid) to fulfil the minimal requirements for the formation and function of the platform at the mitochondrial membrane. Our results shed light on the active functional role of cardiolipin, bridging the gap between death receptors and mitochondria