Ph.D. Work

Abstract
The morphological changes of lipid membranes upon interaction with an amphiphilic polymer were studied experimentally and theoretically and various shape instabilities were observed. My study focused mainly on the pearling of hollow cylinders, the coiling of multilamellar tubes and the budding and fingering of highly oblate vesicles. A single model, based on the induction of local curvature by the polymer, is shown to agree with all the observed phenomena.

Experimental results

The experiment
As a first step towards describing biological membranes, we study a simplified model system composed of a lipid membrane interacting with an amphiphilic polymer. The polymer (yellow) inserts its hydrophobic side-groups (red) into the membrane (black).

Pearling
Snapshots of a multilamellar tubular vesicle of SOPC undergoing a pearling instability: (a) 0, (b) 70 sec. and (c) 150 sec. after the onset of pearling. (d) Image of an inhomogeneous pearled structure at late stages of the instability, 900 sec. after the onset of pearling. The polymer concentration is much larger than in (a-c). The scale bars represent 20 mm.

I. Tsafrir, D. Sagi, T. Arzi, M.A. Guedeau-Boudeville, V. Frette, D. Kandel,  and J. Stavans, Pearling instabilities of membrane tubes with anchored polymers, Phys, Rev. Lett, 86,1138-1141, 2001.

Budding
Buds form at the rim of highly oblate vesicles. This is probably due to the fact that the rim has the greatest curvature. At low polymer concentrations, the rim displays enhanced fluctuations (a)-(b). Nucleation of a bud suppresses rim fluctuations (c). The bud grows into a tube (d-f). The scale bar represents 10 mm.

Alternatively, flushing the cell with polymer solution of large concentration, induces the simultaneous formation of buds all along the rim (b). Buds grow into tubes which subsequently pearl (c). At high polymer concentrations on the membrane, small structures of high curvature result (d). The scale bar represents 10 m.

Local injection leads to local budding

I. Tsafrir, Y. Caspi, T. Arzi, M.A. Guedeau-Boudeville,  and J. Stavans, Budding and Tubulation in Highly Oblate Vesicles by Anchored Polymers, Phys, Rev. Lett, 91, 138102 (2003)

Coiling
The cylindrical multilamellar tubes, called myelin figures, consist of a large number of membrane bilayers reaching almost to the core. Due to the large number of concentric layers, these structures cannot pearl, instead, when hydrated with polymer they bend into coils. This system is unique in that coils form in the absence of twist.

V. Frette, M.A. Guedeau-Boudeville, I.Tsafrir, L. Jullien, D. Kandel, and J. Stavans., Coiling of cylindrical membrane stacks with anchored polymers, Phys, Rev. Lett, 83, 2465-2468, 1999.

I.Tsafrir, M.A. Guedeau-Boudeville, D.Kandel, and J.Stavans, Coiling instability of multilamellar membrane tubes with anchored polymers, Phys, Rev. E, 663,031603, 2001.

Theory

The model
The proposed model describing the system is based on the following assumptions: Membrane curvature and polymer concentration are coupled. The polymer induces a local spontaneous curvature H0. Polymers can diffuse on the membrane. The membrane is modeled as  two-dimensional lattice, where each site can be occupied by at most one polymer molecule. Microscopic Hamiltonian:

where a is the area of a site, k is the bending modulus of a lipid site, sij is 1 for a polymer site 0 otherwise, k' is the bending modulus of a polymer site, Hij is the curvature of site and H0 is the spontaneous curvature.

Pearling
Analysis of the instability near threshold, revealed that the ratio between the radius of the tube and the wavelength of the instability, 2pR0/P is characteristic of spontaneous curvature.

When the polymer is introduced locally, by using a micropipette, the instability propagates from the region where the polymer is added. When the pipette is removed, polymer continues to diffuse along the tube. The pearls open up sequentially. Two theoretical mechanisms, ADE and SC, were shown to be responsible. The timescale associated with ADE is roughly 50 times faster than the one associated with SC.

Coiling
The equilibrium state of a tube is calculated as follows: Pay energy for bending a tube, keeping polymer distribution homogeneous. Gain energy from inhomogeneous polymer distribution. Pay for loss of mixing entropy. The energy gain from an inhomogeneous polymer distribution depends linearly on H0, while the bending energy and the entropy do not . Thus for large enough values of H0 the free energy of an inhomogeneous bent tube is larger than that of a homogenous straight one.

The curvature at every point is determined by the center line:

The free energy of a thick tube with constant centerline curvature, after integrating over the entire stack, is:

This energy predicts that below a critical polymer concentration the equilibrium shape is that of a straight tube, while above it the tube form maximally curved structures.

Geometrical phase diagram for double helices, r0 is the outer radius, 2ph is the pitch of the central line, and r2 is the radius of the smaller central line.

Some comparisons between the theoretical models and actual observed coils: