SYNOPSIS OF

Eect of in-plane order and activity on

vesicular morphology
A THESIS
to be submitted by
N. RAMAKRISHNAN
for the award of the degree
of
DOCTOR OF PHILOSOPHY
DEPARTMENT OF PHYSICS
INDIAN INSTITUTE OF TECHNOLOGY MADRAS.
August 2011
1 Introduction
Lipids are one amongst the four building blocks of biology, with the other three being
amino acids, nucleic acids and sugar molecules[1]. These fatty acids, which are carboxyl
group containing hydrocarbons, are the most abundant molecules in the cell, numbering
over a thousand in both eukaryotes and prokaryotes [2]. From a biological point of view,
supramolecular organization of lipids have low functionality compared to biopolymers like
proteins and DNA/RNA, which are poly-amino acids and poly-nucleotides respectively. A
lipid molecule can either be polar or apolar depending on the chemical moieties attached
to the carboxyl group. Apolar lipids, in a polar solvent like water, aggregates into lipid
droplets that are the known to be the energy store of a cell [3] and are more interesting
from a functional point of view.
The structurally important, polar lipid molecules are characterized by a hydrophilic
part called the head and a hydrophobic chain called the tail. This is illustrated in Fig.1(a)
and (b) for the case of a DMPC lipid. When in an aqueous environment, the amphiphilic
lipids, driven by hydrophobic forces, spontaneously aggregates into a structural entity
called membrane. The simplest known membrane is a lipid bilayer, see Fig.1(c) and
(d), in which the lipid molecules are organized in a manner such that the oily tails are
shielded from the solvent by the hydrophilic head groups. Biological membrane denes the
boundary and shape of every existing cell and its organelles. These omnipresent structures
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(c) Ilat bilayer
(d) closed bilayer
10nm
Hydrophilic
(a) 14:0 dimyristoyl-phosphodylcholine
Hydrophobic
(b)
Head
Tail
Figure 1: A lipid molecule and some of its self organized structures. Shown are (a) the
chemical composition of a DMPC lipid, with the hydrophobic and hydrophilic
regions shaded dierently, (b) the representative model of a lipid, with head
and tail groups marked, (c) a cross section of a membrane bilayer and (d) a
closed bilayer.
exist in a variety of conformations ranging from simple spherical, as the plasma membrane
1
enclosing the cytoplasm, to complex convoluted shapes, seen in case of cell organelles like
Endoplasmic Reticulum(ER) and Golgi.
In this thesis, with the use of computational tools and phenomenology based physical
models we attempt to throw some light on one of the most intriguing questions of biology
: How does a cell membrane generate and stabilize curvature ?.
1.1 Membranes - A physicist description
Being self assembled structures, with no chemical links between the constituent molecules,
membranes are a class of soft materials, which have low energy densities with thermal
energy k
B
T being a relevant energy scale [4]. The thermally induced membrane shape
uctuations are pronounced enough to be visible under a microscope, which is a rst
indication of its softness. A membrane respond to changes in its physical and chemical
environment with a smooth change in its shape, with features reminiscent of an elastic
sheet. Further, as revealed by single molecule tracking experiments[5], membranes are
uid surface and hence do not support tangential shear. The self assembled nature of a
membrane makes it tensionless, a key feature that distinguishes it from other thin shell
systems, like a soap bubble.
Biological membranes have widely separated length scales and time scales. It measures
a few nanometers in the transverse direction and extends to the order of microns laterally.
Similarly, the timescale of in-plane diusion is much faster ( s) compared to that of
lipid translocation between the monolayers, which ranges between hours to days. In this
limit, a lipid bilayer can be represented as a thin, exible, uid sheet, of constant area
at length scales matching its maximum dimension. The sheet represents the mid plane
of the membrane since the structural behaviour of the individual monolayers, coupled to
each other by the strong hydrophobic association among their lipid tails, closely follows
the midplane. The thermodynamic properties of this 2D uid sheet, henceforth be called
the membrane, embedded in three dimensional space is given by the elastic energy,
H
sur
=

dx
1
dx
2

2
(H C
0
)
2
+
G
R

dV, (1)
due to Canham and Helfrich [6, 7]. The surface of the membrane is dened by its internal
2
coordinates [x
1
, x
2
] and the chemistry of the lipid molecules are recast in terms of the
phenomenological parameters the bending rigidity and
G
the Gaussian rigidity. H and
R are the gauge invariant measures dened on the membrane surface, namely the mean
and Gaussian curvatures. C
0
is the spontaneous curvature induced in the membrane due to
a multitude of reasons like lipid geometry, asymmetric lipid distribution in the monolayers,
etc.. A uid membrane is semi-permeable since it allows only selected molecules to diuse
through it. Such a barrier for a closed membrane surface, with volume V , results in an
osmotic pressure dierence, p = p
in
p
out
, between the interior and exterior of the
membrane. Within this model, the equilibrium conformations of a membrane, with xed
topology, is entirely determined by the paramters and p. A detailed review of various
physical models and properties of uid membranes can be found in [4].
1.2 Triangulated surfaces : Discrete model for membranes
The scope of the continuum approach is limited to perturbation studies of surfaces that
can be parameterized as a single valued function of the inplane coordinates (x
1
, x
2
). As an
alternative, computational approach has emerged as a powerful tool to probe membrane
properties in the non-perturbative limit. Among all discrete representations of continuous
surfaces, the triangulated surface model is best suited for modelling macroscopic mem-
branes. A triangulated surface is constituted of T triangles (faces) connected through
N vertices (nodes), with each triangle representing a at region of the membrane. The
topology of the surface is determined using its Euler characteristics, = 2(1 g) h,
where g and h are the number of handles and holes in the surface. The number of faces,
nodes and edges together dene the topology of the surface as, = N + T L. The
elastic energy of the membrane sin the discrete limit is given by,
H
sur
=
N

v=1

c
1
(v) +c
2
(v)
2
C
0

2
A(v) p V. (2)
The geometry of a membrane is dened by the local surface quantiers namely the prin-
cipal curvatures, c
1
(v) and c
2
(v), and the principal directions. These quantities, dened
on the vertices of a triangulated surface, are computed using techniques introduced in
our article [8]. Chapter 2 of the thesis will contain all details pertaining to simulations
3
using triangulated surfaces and will lay the framework for its applications described in
later chapters.
1.3 Anisotropy in membranes
The equilibrium solutions of Eqn.(1), representing an isotropic membrane, are minimal
surfaces that rules out the occurrence of any of the highly curved shapes found in bi-
ological systems and described earlier. This underlines the fact that the properties of
the membrane are dependent on some parameters that have been integrated out in the
process of coarse graining. Anisotropy in the elastic behaviour of membranes is one such
parameter, which has been recognized for long to inuence membrane shapes[9].
A membrane exhibits anisotropic behaviour when it develops an internal local order,
due to constraints on its molecular degrees of freedom in the form of lipid tilt, lipid rota-
tion about its own axis and chirality[10]. The origin of anisotropy can also be external,
as seen in the case of membranes interacting with external macromolecules like proteins.
Recent experiments have identied a large set of peripheral membrane proteins, like BAR
domain containing proteins[1114], Dynamin superfamily proteins [15], Nexins, epsins,
reticulon containing Dp1/Yop1[16] etc., to spontaneously deform a membrane. In addi-
tion, anisotropy can also arise from the geometry of lipid molecules, as observed in the
case of the dimeric head Gemini surfactant [17].
In our studies, the anisotropic local order is treated as a vector eld conned to
the tangent plane of the membrane. Emergence of highly curved and biologically rele-
vant membrane shapes due to the collective eect of an achiral nematic in-plane eld is
demonstrated in Chapter 3. In addition, the importance of topological defects and their
role in driving membrane tubulation has also been detailed.
1.4 Activity : a source of membrane remodelling
Budding and fusion of vesicles are membrane mediated transport mechanisms involved
in cargo intake and waste segregation, that are utterly essential to maintain cellular
compositions suitable for its functionality. These mechanisms, wherein small transport
4
vesicles encapsulating the cargo fuses or pinches o from a at membrane, also play an
important role in the exchange of material between cell organelles. Fission and fusion of
vesicles leads to large scale remodelling in the shape of membrane, primarily driven by
protein machineries. All membranous structures in a cell are subjected to various levels
of material ux, hence the observed shapes of cell organelles are also dependent on the
ux generated active forces. With the ssion and fusion being non equilibrium events,
driven by ATP hydrolysis, it would be instructive to treat the shapes of most transport
intensive cellular structures as steady states rather than at equilibrium.
The steady state nature of cell organelles is well demonstrated in the ER-Golgi secre-
tory pathway. The Golgi, an interconnected stack of membranes, is subjected to inbound
and outbound transport vesicles from and to the Endoplasmic Reticulum(ER). It frag-
ments into spherical and tubular vesicles in the mitotic phase[2], of cell division, during
which all usual interphase cell functions including intracellular transport are inhibited.
The dependence of Golgi structure on the vesicular transport has also been demonstrated
in many other experiments. We investigate the role of the bidirectional transport in shap-
ing closed membrane compartments using a polymer loop, the 2D version of a membrane,
in Chapter 4. As expected, interesting shapes, with equally interesting statistics, are
stabilized when material is added and removed in a non-equilibrium manner.
The Endoplasmic reticulum (ER) serves as the entry point to many macromolecules,
like peptides and water soluble proteins, destined to cell organelles or to the plasma
membrane. A cytosolic protein, for instance, remains attached to the rough ER while
being maturated by membrane bound ribosomes. On maturation, it diuses around
and escapes into the ER lumen through a translocator protein. Interactions of these
macromolecules, that bind and unbind to the membrane in a stochastic manner, induce
local membrane deformations vital to ER morphogenesis. More examples of such protein
induced membrane deformations are also seen in mitochondria and choloroplasts [2].
In chapter 5 of the thesis, this phenomenon is investigated with the aid of a model for
protein binding and unbinding activity, on a triangulated surface. ER relevant membrane
shapes like discs and tubes are the natural steady states of this model. It has also been
shown that active forces tend to renormalize the osmotic pressure of a vesicle, resulting
in an enhanced deating pressure.
5
2 Motivation, Objective and Scope
Biologist have started to view lipid membranes as a vital component for cell function and
the view of it as the dull passive barrier, not important for cell function, has changed.
Emerging experimental evidences indicate the role of membranes in many biological events
as a barrier, carrier and target [1]. It has been found, of late, that a large class of proteins,
called curvactants, readily remodels the shape of the membrane they are attached to.
The recruitment of these proteins and the subsequent deformation of the membrane are
precursor to many endocytic events. The mechanisms of membrane moulding, by BAR
domain containing proteins, have been investigated in many microscopic and mesoscopic
studies[18, 19]: all of which have highlighted the role of anisotropy.
Our interest in membranes with anisotropic elastic behaviour stems from the following
reasons : (a) To develop a macroscopic model that accounts for the dierent classes of
curvactant proteins and also the resulting membrane shapes, (b) anisotropic membranes
being paradigm systems for many other classes of soft matter, like nematic shells, ne-
matic elastomers, etc., a study of these surfaces can be extremely useful in investigating
their properties, (c) to understand the structure and dynamics of topological defects on
deformable surfaces, of which little has been known.
Disassembly of Golgi apparatus on inhibition of vesicular transport, thus establish-
ing the steady state nature of transport intensive cell organelle shapes, has been known
for long. But,the number of studies that account for the eect of this activity on mem-
brane shapes are minimal. We make an attempt, in this thesis, to look into the problem
of membranes subjected to non-equilibrium activity arising from the material transport
and also from the stochastic binding and unbinding of curvactants, generally seen in the
Endoplasmic Reticulum.
6
3 Summary of the Research Work
3.1 Random surface with and without in-plane order
Discretized uid membranes are modelled as triangulated surfaces, with its elastic energy
given by Eqn.(2). Investigations into the anisotropic behaviour of membranes, described
in Sec.1.3, requires the complete description of the triangulated surface, in terms of its
principal curvatures and directions. Starting with techniques to determine the surface
quantiers, chapter 1 lays down the computational framework used in all problems de-
scribed later.
To characterize the simulation scheme, we rst look at the isotropic membranes . The
results obtained here are qualitatively similar to that of earlier studies[20, 21]. Here,
absence of a crumpled phase, for 0, is characterized by linear scaling of the radius of
gyration, R
2
G
N. The specic heat, computed from the uctuations of the elastic energy
given by Eqn.(2) and shown in Fig.2(a), shows a peak that saturates in the asymptotic
limit, with values

and C
max
being 4.4 and 1.4 respectively. The presence and saturation
of the peak indicates a smooth transition between the branched polymer and extended
phase of the membrane. Persistence length of the membrane, calculated from nite size
scaling of the volume, shows an behaviour of the form exp(). We observe, as in Fig.2(b),
a low kappa regime with = /6 for < 3.0 that crosses over to a higher value of , as
expected by theory, for biologically relevant values of (10 20k
B
T).
0 2 4 6
0
0.5
1
1.5
77
110
245
302
677
2030
2702
3677
0 0.002 0.004
1
2
3
4
1
2
3
4

C
(
N
,

)
N
1
C
m
a
x

(a)
0 1 2 3 4 5
1
10
100

p
exp

(b)
Figure 2: (a) Specic heat C(N, ) versus for varying N. The position of C
max
(N, )
(circles tted with solid line) and

(N)(squares tted with dotted line) are

shown in the inset. (b)Persistence length,
p
, as a function of
We then come to membranes with anisotropic elasticity. Local order developed in the
7
membrane is represented as a vector eld dened at the vertices of the triangulated surface
as m(v) = cos (v)

t
1
+ sin (v)

t
2
, where

t
1
and

t
2
are the principal directions at vertex v
and (v) the angle between the in-plane eld and

t
1
. A nematic eld in the ordered phase,
with self interactions given by the Lebwohl-Lasher Hamiltonian [22], stabilizes four +1/2
disclinations when ordered on a surface of spherical topology of which one is shown in
Fig.3(a). When on a non-deformable sphere the defects arrange themselves in a baseball
(a) (b)
(c)
Figure 3: (a) A +1/2 disclination is shown, (b) the baseball texture of a nematic eld on
a rigid sphere, with the shaded regions marking the defect core, (c) a deformed
surface with the defects on the vertices of a tetrahedron, for J
2
= 5k
B
T.
texture, see Fig.3(b), while a deformable surface take shapes that preserve the tetrahedral
arrangement of defects, as in Fig.3(c).
3.2 Nematic membranes
Most of the observed curvature inducing membrane proteins have nematic symmetry.
While some of these proteins could have an explicit chiral center they are not observed
to induce chiral deformations on the membrane. For reasons of biological relevance and
simplicity, we consider only achiral nematic membranes with anisotropic bending energy
of the form,
H
anis
=
N

v=1
A(v)

[H

(v) c

0
]
2
+

[H

(v) c

0
]
2

. (3)
This model is studied in detail in Chapter 2. Here,
()
and c
()
0
are the direc-
tional bending rigidities and spontaneous curvature parallel(perpendicular) to the eld
m. H

(v) = c
1
(v) cos
2
(v) +c
2
(v) sin
2
(v) is the directional curvatures of the membrane
along m. A variety of shapes can now be obtained as the directional spontaneous curvature
and the stiness is varied. Fixing = 10,

= 5,

= 0 and J
2
= 3, all in units of k
B
T,
8
the obtained membrane conformations as a function of directional curvature C

0
are shown
in Fig.4. In the absence of any stiness-orientation coupling, the +1/2 disclinations have
(a) Oblate (b) Tubular (c) Disc (d) Inner tubes
(e) Tubular (f) Corkscrew (g) Branched (h) Branched
Figure 4: Membrane conformations for C

0
= 0.0 (a), -0.2(b), -0.4(c), -0.6(d), 0.2(e),
0.4(f), 0.5(g) and 0.6(h) with = 10,

= 5,

= 0 and J
2
= 3.
a repulsive interaction leading to them being equally spaced, shown as a single peak in the
measured distribution of geodesic distances between the defects in Fig.5(a). Directional
bending stiness (

= 0) alters this distribution considerably, since curvature inuences

the interaction between defects[23]. When C

0
> 0 two of the +1/2 defects move towards
each other to form pairs and the vesicle deforms into tubes: these are shown in the lower
row of Fig.4. The short distance peak, in Fig.5(a), for C

0
= 0.2 0.4 corresponds to the
defect pairs at the end of the tube. The equilibrium value of the principal curvatures is
set by C

0
, a distribution of which is plotted in Fig.5(b). As illustrated in panel III of
Fig.5(b), when

= 0 and C

0
> 0 the maximum principal curvature takes values such
that c
1
C

0
as 0. Tubes and discs are stabilized for small and intermediate values
of C

0
< 0, but the self avoidance constraint inuences the shape of membranes with large
negative directional spontaneous curvature. Caveola like structures, shown in Fig.4(d),
are a result of this constraint and these shapes are characterized by a bimodal distribution
of the minimum principal curvature, c
2
.
9
0 25 50 75 100 125
0
0.01
0.02
0.03
0.04
0.05
Separation between
far away defects.
Defects at equal distance
on a tetrahedron.
Geodesic distance,
P
(

= 0 ; C

0
= 0

= 5 ; C

0
= 0.2

= 5 ; C

0
= 0.3

= 5 ; C

0
= 0.4

= 5 ; C

0
= 0
(a)
0
0.005
0.01
-1 -0.5 0 0.5 1
0.2
0.5
0.7
0
0.005
0.01
-0.2
-0.5
-0.7
-1 -0.5 0 0.5 1
0
0.005
0.01
(I)
(II)
(III)
c
1
&c
2
P
(
c
1
)
P
(
c
2
)
P
(
c
1
,

)
= 5
= 10
= 20
(b)
Figure 5: (a) Distribution of the geodesic distance between the defects. (b) The equi-
librium statistics of minimum (c
2
) and maximum (c
1
) principal curvatures, are
shown in panel I and II respectively. It can be seen from panel III that c
1
C

0
as 0.
Defect dynamics play an important role in remodelling membranes. We have identied
a new mechanism of membrane tubulation wherein proliferation of additional defect pairs
drives a spherical membrane into a tube. The process of tubulation with all intermediate
steps is illustrated in Fig.6.
e) Defect pair d) Unlike defect repel c) Defect Proliferation b) Deformation a) Sphere with
isotropic nematics by thermal fluctuations of membrane leading to tube formation annihilation
+1/2 and 1/2
defect pair
Figure 6: Kinetics of the spiral formation driven by proliferation of additional defect
pairs.
3.3 Active membranes in two dimensions
A ring polymer represents a cross section of a vesicle. Modelling the self avoiding ring
polymer as beads, of diameter a, interconnected by tethers, with length a < l <

3a, we
study the eect of active forces, resulting from the membrane mediated material transport.
Activity is introduced in the model by adding and removing beads in a non equilibrium,
curvature dependent manner.
As shown in Fig.7(a), a semi-exible polymer ( 120) when subjected to activity of
10
2 5 10 15 20 25 30
2
5
10
15
20
25
30
2 5 10 15 20 25 30
2
5
10
15
20
25
30

add
r
e
m

E
x
t
e
n
d
e
d

S
h
a
p
e
s
Collapsed looped structures
(a)
0 200 400 600 800
0
0.0005
0.001
0.0015
-2 -1 0 1 2
10
-4
10
-3
10
-2
Geometric distance, r(s)

(
0
)

(
r
)

= 0 ; Inactive
= 120 ; Inactive
= 120 ; Active
q

(b)
Figure 7: (a) The phase diagram of a semi-exible polymer in the
add
-
rem
parameter
space, for = 120 and activity rate

= 0.1N/MCS. (b) Density-density

correlation shows that an active polymer is more compact than a self avoiding
random walk. Data shown for
add
= 30 and
rem
= 10.
addition and removal of monomers stabilizes into a looped structure above some values
of the chemical potentials to add (
add
) and remove (
rem
) a bead from a region of unit
curvature. These looped structures are true steady states, which is conrmed by their
reversal to circular structure on inhibition of activity. As is evident from the density
density correlation (0)(r), in Fig.7(b), the looped polymer is more compact compared
to a self avoiding random walk, that leads to spatial proximity of beads far separated
along the contour of the polymer.
Two beads separated by s along the contour are said to be in contact when their
geometric distance is such that 1 < r(s) <

3. An estimate of the contact probability

for a self avoiding walk (SAW) and an active semiexible polymer are given in Fig.8(a).
The observed value P(r) s
2.80.2
, for a SAW, is very close to the theoretical estimate
of s
2.6875
[24]. The active polymer, on the other hand, shows a linear regime for s < 40
and exhibits a power law decay with P(r) s
1.550.1
for higher values of s. This scaling
behaviour holds for all looped structures stabilized at dierent values of
add
and
rem
.The
presence of activity sets a length scale in the polymer that is reected in the loop size
distribution, given in Fig.8(b). The loop length shows a non Gaussian distribution, peaked
at s 40, and is invariant under change in system size.
The steady state shapes of two dimensional vesicles or ring polymers, arising out of
active addition and removal of monomers, is shown to stabilize highly curved shapes,
usually associated with transport intensive organelles like the Golgi. Further, the contact
statistics of the active polymers closely resemble the hierarchical structures of DNA or-
ganization in chromatin, believed to be generated by binding of histone proteins. Hence
11
10 100 1000
1e-06
1e-05
0.0001
0.001
0.01 5-5
5-10
10-5
10-10
20-5
Random walk
Contour length, s
P
(
1
<
r
(
s
)
<

3
)
s
2.80.2
s
1.550.1
(a)
0 20 40 60 80 100
Contour length, s
0
0.01
0.02
0.03
0.04
L
o
o
p

l
e
n
g
t
h

d
i
s
t
r
i
b
u
t
i
o
n
,

P
(
s
)
2500
5000
10000
Average polymer size
(b)
Figure 8: (a) Contact probability distribution for a SAW and active an polymer. (b) The
polymer size invariant, non-Gaussian, distribution of loop length.
active polymers, described in Chapter 4 of this thesis, can also be used a model system
to understand the organization of histone bound DNA.
3.4 Activity induced remodelling of membrane in three dimen-
sions
Membrane deformations caused by the constant ux of water-soluble proteins and the
resulting macroscopic organization has been investigated in Chapter 5. We use a semi-
grand ensemble model with a triangulated surface for the membrane and an Ising variable,
dened on its vertices, denoting protein bound ( = 1) and depleted ( = 1) regions.
This two state curvature active eld {} is swapped from = 1 = 1 when a
protein unbinds from the membrane and vice-versa, in a non-equilibrium manner at a
dened activity rate

. The energy of such a membrane, with N

1
bound ( = 1) and
N
1
unbound ( = 1) sites, is given by,
H
active
=
N

v=1

H(v)
(1 +
v
)
2
C
0

2
p V J
N

v=1

v
, (4)
where J is the strength of protein self interaction. The shapes of the membrane with the
steady state active eld composition N
1
= 0.1N, at a xed rate of binding(unbinding)
activity

= 0.1N/MCS and = 120, is studied as a function of the spontaneous curva-

ture C
0
, in Fig.9. The spherical shape of the membrane is stable to small deformations
induced by binding and unbinding of the active eld for spontaneous curvature in the
range C
0
= 0.0 0.6. The eect of activity is seen at higher values of C
0
as the steady
12
state membrane conformations change to tubular and disc like. Such systematic changes
in shapes are also observed by varying N
1
or

.
(a) Spherical (b) Tubular (c) Disc & tubes
Figure 9: Steady state shapes of an active membrane for (a) C
0
= 0.0 0.6, (b) C
0
=
0.6 0.85 and (c) C
0
= 0.9 with J = 0.0, = 20, N
1
= 0.1N and

=
0.1N/MCS. Speckled regions indicate the spatial locations of the active eld.
Analysis of these transformations indicates that the volume undergoes a sharp transi-
tion, as shown in the main plot of Fig.10(a), when the membrane goes from a spherical to
other non-trivial shapes. With the surface area of the membrane remaining nearly con-
stant, its aspect ratio changes with change in the enclosed volume. The range of shapes
are qualitatively similar to those predicted theoretically for dierent aspect ratios. Activ-
ity on a membrane manifests as an additional osmotic pressure dierence p
R
< 0, which
in turn drives the volume change in the membrane. The renormalization of pressure for
dierent values of

and C
0
are shown in Fig.10(b), with the width of the lower plateau
being a measure of p
R
.
0 0.5 1 1.5
2600
2800
3000
3200
0 0.5 1 1.5 2
5e+03
1e+04
2e+04
Quasi Spherical
Tubes, discs and Stomatocytes

C
0
C0
= 0.1 N/MCS
= 0.15 N/MCS
= 0.2 N/MCS
= 1.0 N/MCS
(a)
0 2 4
0.5
0.6
0.7
0.8
0.9
1
Sphere of Volume V
0
p
0
V
(

,
C
0
)
V
0
= 0.1 N/MCS
= 0.0 N/MCS
= 10 N/MCS
= 1 N/MCS
= 5 N/MCS
(b)
Figure 10: (a) Average volume (main) and area (inset) of an active vesicle as a function
of C
0
for = 20, J = 0.0 and N
1
= 0.1N. (b) Rescaled volume of a membrane
subjected to activity as a function of bare osmotic pressure p
0
for = 20,
C
0
= 0.8, J = 0.0 and A
0
= 0.1N.
Studies of membrane with spatially localized activity (data not presented here) shows
that the active elds are conned to the highly curved regions of the membrane while the
inactive regions remain at. This is a conclusive proof implicating the role of active forces
in generating highly curved membrane shapes.
13
4 Conclusions
This thesis looks into the various aspects of membrane morphogenesis from a physical
point of view. In particular, the role of anisotropy, due to a local order in the membrane,
and active forces, arising from material transport and binding and unbinding of proteins,
have been looked into in detail. The key achievements are as follows,
1. Computational techniques to handle vector elds on membrane surface has been
introduced.
2. Presence of anisotropic elastic behaviour with a nematic symmetry is shown to
drive the membrane into biologically relevant shapes like tubes, branches, discs and
caveola like. Further, the proliferation and dynamics of topological defects denes
a new mechanism for membrane tubulation.
3. A ring polymer subjected to non-equilibrium curvature dependent activity stabi-
lizes compact looped structures. The contact probability as a function of contour
length scales as s
1.550.1
for an active polymer compared to s
2.80.2
for a self avoid-
ing walk. Activity denes a length scale in the polymer which is reected in the
invariance of the loop size distribution.
4. It is shown that, stochastic binding and unbinding dynamics remodels a uid mem-
brane into non-trivial shapes. This activity manifest as a deating pressure that
drives the conformational change.
To conclude, we point out that anisotropic elastic behaviour and active forces can be
key parameters stabilizing highly curved membrane morphologies. Hence, they should be
an integral part of any theory aiming to explain the morphogenesis of biological mem-
branes.
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5 Proposed Contents of the Thesis
The outline of the thesis is as follows:
Chapter 1 Introduction to membranes
Chapter 2 A model for uid surfaces with and without in-plane order
Chapter 3 Nematic membranes
Chapter 4 Active membranes in two dimensions
Chapter 5 Active membranes in three dimensions
Chapter 6 Conclusions
6 Publications
6.1 Papers in Refereed Journals
1. Monte Carlo Simulations of vesicles with in plane orientational order
N. Ramakrishnan, P. B. Sunil Kumar, John H. Ipsen
Phys. Rev. E, 81, 041922(2010)
2. Modelling anisotropic elasticity of membranes
N. Ramakrishnan, P. B. Sunil Kumar, John H. Ipsen
Macromolecular Theory and Simulations, DOI: 10.1002/mats.201100002, 2011
6.2 Presentations in Conferences
1. N. Ramakrishnan, P. B. Sunil Kumar, John H. Ipsen
Poster entitled In-plane order and membrane morphology presented at
School and Conference on Mathematics and Physics of soft and Biological matter, The
Abdus Salam ICTP, 2-13 May 2011
2. N. Ramakrishnan, P. B. Sunil Kumar, John H. Ipsen
Poster entitled Anisotropic elasticity of uid membranes presented at
Novel Simulation Approaches to Soft Matter Systems, MPI Dresden, Germany, 20 - 24
Sep, 2010.
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