2710 Biophysical Journal Volume 106 June 2014 2710–2719

Drag of the Cytosol as a Transport Mechanism in Neurons

Matan Mussel,† Keren Zeevy,† Haim Diamant,‡ and Uri Nevo†§*

†
Department of Biomedical Engineering, Tel Aviv University, Tel Aviv, Israel; ‡School of Chemistry, Tel Aviv University, Tel Aviv, Israel; and
§
Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel

ABSTRACT Axonal transport is typically divided into two components, which can be distinguished by their mean velocity.
The fast component includes steady trafficking of different organelles and vesicles actively transported by motor proteins.
The slow component comprises nonmembranous materials that undergo infrequent bidirectional motion. The underlying mech-
anism of slow axonal transport has been under debate during the past three decades. We propose a simple displacement
mechanism that may be central for the distribution of molecules not carried by vesicles. It relies on the cytoplasmic drag
induced by organelle movement and readily accounts for key experimental observations pertaining to slow-component trans-
port. The induced cytoplasmic drag is predicted to depend mainly on the distribution of microtubules in the axon and the organ-
elle transport rate.

INTRODUCTION
Neurons are highly polarized cells with axons capable of
extending long distances. Individual axons are microscopic
in diameter (typically of the order of 1 mm), whereas their
lengths may span as much as one meter in humans (e.g.,
axons in the sciatic nerve and spinal cord) and even more
in large animals such as giraffes or whales. Since axons
are cellular structures with a unique morphology, the
morphogenesis, integrity, and function of neurons are partic-
ularly dependent on active intracellular transport, also called
axonal transport (1,2).
There are three known major constituents of the neuronal
cytoskeleton: microtubules, microfilaments, and neurofila-
ments (2). In the axon and dendrites, microtubules and
neurofilaments are the major longitudinal cytoskeletal
filaments. In the synaptic regions, such as the presynaptic
terminals and postsynaptic spines, microfilaments and
microtubules form the major cytoskeletal architecture (3).
There are also microfilaments along the axon, which mainly
provide a scaffold beneath the axonal plasma membrane and
around microtubules (4,5).
The lines of transport, communication, and control in the
cell are provided by microtubules. These dynamic polymers
create a network of tracks for motor proteins that carry
various vesicles and organelles (referred to hereafter as
cargoes), with diameters of the order of 10–100 nm (6).
Microtubules appear as bundles of tubes dispersed across
the axon and are aligned mostly along the axon axis to allow
long-range transport (see, e.g., Fig. 8-1 in Siegel (2)). In
comparison to the two other intracellular cytoskeletal
filaments, microtubules are the most rigid, thus ensuring
mechanical stability under the forces of large cargo
movement (7).
Submitted January 7, 2014, and accepted for publication April 28, 2014.
*Correspondence: nevouri@tau.ac.il
Matan Mussel and Keren Zeevy contributed equally to this work.
Editor: David Odde.
Ó 2014 by the Biophysical Society
0006-3495/14/06/2710/10 $2.00
Axonal transport plays a few major roles. 1), It is respon-
sible for bidirectional molecular signaling over long dis-
tances. 2), It provides essential cellular organelles and
materials from the cell body to the entire axon (anterograde
transport by the kinesin motor protein). This supply of
newly synthesized proteins and lipids to the distal synapse
maintains axonal activity. 3), It causes molecules destined
for degradation to return from the periphery to the lyso-
somes in the soma (retrograde transport by the dynein motor
protein) (8,9). The modes of transport through the axon are
typically divided into two categories: a fast component and
a slow component (1). The components are distinct in their
mean velocity, as measured in studies of radioactively
labeled proteins (10–15).
The fast component is a directed movement of mem-
branous cargoes. Small cargoes move continuously in one
direction and maintain almost uniform velocity that ranges
between 0.4 and 3 mm/s, whereas larger cargoes exhibit
irregular saltatory motion (16–21). The fast transport is
carried primarily by molecular motor proteins that bind
directly to the cargo surface (22–24). The directionality of
the cargo depends on the motor type and the polarity of
the microtubule tracks (1).
The slow component (SC), which is the focus of this work,
is usually divided into two subgroups with distinct overall
rates: SCa and SCb (1). SCa transports cytoskeletal proteins,
whereas SCb transports hundreds of different proteins that
are important for synaptic and axonal maintenance,
including metabolic enzymes, chaperons, presynaptic pro-
teins, and cytoskeletal proteins (see Lasek and colleagues
(25,26), Baitinger and Willard (27), Nixon et al. (28), Yuan
et al. (29), Bourke et al. (30), and Roy et al. (31) and refer-
ences therein). Key experimental observations concerning
both SCa and SCb are as follows (31–35). 1), Transported
soluble molecules spend most of their time pausing, and their
movement appears bidirectional. 2), They are not attached to
vesicles but homogeneously distributed throughout the entire
http://dx.doi.org/10.1016/j.bpj.2014.04.037
Cytosol Drag in Neurons
cytoplasm. However, their transport was found to depend on
motor proteins (35). 3), They move at broadly distributed
velocities, ranging from close to zero to the velocity of the
fast component, i.e., a few mm/s. 4), Slow-component pro-
teins were even observed to move side by side with fast-
component cargoes (34).
In contrast to the transport mechanism of the fast compo-
nent, that of the SC is still under debate (36). Blum and Reed
(37) suggested a set of interactions between soluble proteins
and motor proteins. When bound to the motor, the soluble
proteins move, and when not bound, they are stationary.
Additional stochastic models have also been suggested
(38–41) in which soluble proteins bound to motors move
in the cytoplasm at different discrete velocity states and in
which the transition from motion to halt is a probabilistic
one. Scott et al. (35) proposed that soluble proteins move
in the cytoplasm through both passive diffusion and a direct
interaction with hypothetical mobile units. In that model,
the assembly and disassembly from the mobile units is prob-
abilistic. Miller and Heidmann (36) offered a qualitative
description according to which soluble protein dynamics
is governed by several mechanisms, including diffusion
from high to low concentrations, vesicles dispersing soluble
proteins as they travel, and displacement induced by the
elongation of axons.
In this work, we propose an alternative, simpler, mecha-
nism for SC axonal transport. It relies on the indirect hydro-
dynamic interaction (flow-induced drag) that exists between
soluble particles and actively transported cargoes along the
axon. We show, without further conjecture, that this simple
mechanism is sufficient to account for the most relevant
experimental observations mentioned above. An illustrative
metaphor for the suggested mechanism is that of log driving,
i.e., the transportation of logs down a river. However, in the
case of intracellular drag, the river flows intermittently,
being driven by anterogradely and retrogradely moving
vesicles, rather than by a steady external field (gravity).
Earlier theoretical works addressing cytoplasmic flow
along the elongated cells of Characean algae (42–45) are
worthy of mention here. Although those studies also consid-
ered flows induced by actively moving organelles, their flow
scenarios differ qualitatively in their geometry from that
considered here.
The outline of the article is as follows. In the next section,
we describe the model and its assumptions. Technical
methods used to analyze the model are explained in the
Methods section. In the Results section, we provide detailed
data from the analysis in different scenarios. In the Discus-
sion section, we discuss specific implications of the analysis
for actual axonal transport, as well as further implications.
MODEL
The system under consideration is schematically shown in
Fig. 1.
2711
a b
FIGURE 1 (a) Diagram highlighting key features in axonal cross section.
Black dots represent microtubules going through the plane of the figure and
distributed uniformly over the cross section. One cargo is attached to a
microtubule at the upper right corner of the axon. For more details, see
Bray and Bunge (46). (b) Schematic diagram of the model. Spheres of
radius a move at constant velocity along the axial direction of a cylindrical
tube of radius R and length L.
We model the axon as a rigid, fluid-filled, cylindrical tube
of length L and radius R. The problem is treated using cylin-
drical coordinates r ¼ (r, 4, z), where the tube axis lies
along the bz direction. The cargoes are modeled as rigid
spheres of radius a moving inside and along the tube. Effects
related to shape changes of both the axon and the cargoes, as
well as the internal dynamics inside the cargoes, are thus
neglected. We assume that each cargo moves at its own con-
stant velocity, Ui, parallel to the axis of the tube (21). The
tube is filled with a viscous Newtonian fluid at zero
Reynolds number (47) having viscosity h. This may seem
a crude oversimplification, given the complexity of the cyto-
plasm. However, based on our results, the detailed response
of the fluid should not have a strong effect on the long-range
transport through the tube. We return to this issue in the
Discussion. The hydrodynamics of particles moving
through fluid-filled tubes have been widely studied (see,
e.g., Brenner and colleagues (47–49), Liron and Shahar
(50), and Blake (51)). Here, we focus on the fluid displace-
ment in the Lagrangian framework, i.e., the displacement of
fluid particles resulting from the active motion of the
cargoes. We consider the transport of the SC as a passive
entrainment of soluble proteins by these active flows. This
assumption relies on the fact that the proteins (a few nano-
meters in diameter) are much smaller than the cargoes (with
radius a on the order of 10–100 nm).
Flow problems in such confined geometries are highly
dependent on boundary conditions (52). For example, when
a sphere moves an infinite distance in an unbounded quies-
cent fluid, the net displacement of any fluid particle is infinite
(53). This idealized result arises from the slow 1/r decay of
fluid velocity with distance r away from the moving sphere.
The opposite example is of a sphere moving in a closed tube
with no-slip boundary conditions. The fluid velocity induced
by the sphere decays exponentially with distance (50), result-
ing in a finite displacement of fluid particles.
A key question is the extent of solvent permeation
through the boundaries along the axon. It is known that fluid
diffuses through the membrane, e.g., via aquaporin channels
Biophysical Journal 106(12) 2710–2719
2712
(54). However, how appreciable such permeation is as a
result of cargo movement remains unclear. Hence, we treat
two simplified limits for the boundary conditions to be taken
at the tube edges. 1), For the case of negligible permeation,
we model the axon as a closed tube of a given length, L. 2),
If permeation is significant, we replace the leaky axon with
an open-ended impermeable tube of effective length L
related to the membrane permeability, m. The correspon-
dence between these two systems is found by a simple anal-
ysis of a pressure-driven flow through a permeable tube,
pffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
yielding a decay length of L ¼ R3 =ð8hmÞ. (One can
model the axon more accurately as an actual permeable
tube, yet we prefer to defer such a detailed analysis to a
future study.) We note that the flow patterns in these two
limits are quite different. When a sphere moves in a closed
tube in the axial direction, the fluid is quiescent far away
from the sphere, whereas near the sphere, some fluid trajec-
tories must go against the direction of motion due to mass
conservation (50). By contrast, a similar sphere moving
through an open-ended tube creates a Poiseuille flow far
from the sphere in the direction of motion (55).
An additional property that may influence the fluid
behavior is the friction at the tube walls. For example, obser-
vations in the giant internodal cells of Characean algae
showed that the fluid slides on the inner surface of the cell
(56). Accordingly, slip boundary conditions were used in
modeling that system (43). The appropriate boundary condi-
tions applied by the axonal membrane and the thick protein
layer adjacent to it are unknown. Therefore, we consider the
entire range of possibilities, from the no-slip boundary con-
dition through partial slip to full slip.
METHODS
We numerically solve the Navier-Stokes equations for an incompressible
fluid in the limit of zero Reynolds number (Stokes equations),
Vp þ hV2 v ¼ 0
; (1)
Vv ¼ 0
for a fluid confined inside a tube. In Eq. 1, v(r, 4, z) and p(r, 4, z) are the
fluid velocity and pressure fields. The calculation is done using a finite-
a b
FIGURE 2 (a) Typical trajectories of fluid particles due to the motion of a single cargo along the axis of a cylindrical tube. Backward particle trajectories
are marked in red. (Inset) Enlargement of particle trajectories with positive, zero, and negative net displacement. (b) The displacement profile. (c) Depen-
dence of the fluid displacement on a/R for four radial positions. Physical parameters are a/R ¼ 0.1 (in a and b), L/R ¼ 10. To see this figure in color, go online.
Biophysical Journal 106(12) 2710–2719
Mussel et al.
element software (COMSOL, version 4.2a). For a single sphere moving
at velocity U, the calculation is done in the reference frame of the
sphere, i.e., the sphere is at rest and the fluid flows past it with a far velocity
of U. Trajectories of fluid particles are subsequently calculated by
integrating the velocity field and transforming back to the fluid reference
frame. For several spheres, we employ a superposition approximation after
confirming its validity for a number of spheres moving at the same velocity
(see Results for details).
Equation 1 is Galilean-invariant and, therefore, the same in the fluid and
sphere reference frames. The entire system, however, is not Galilean-
invariant, which is reflected in different boundary conditions. We begin
with the boundary conditions at the edge of the tube. For a closed
tube the boundary conditions are zero fluid velocity, v(r, 4, 0) ¼
v(r, 4, L) ¼ 0, which in the reference frame of the sphere turn into condi-
tions of constant velocity, vðr; 4; 0Þ ¼ vðr; 4; LÞ ¼ Ubz . For an open-
ended tube, the boundary conditions at the edges, in both reference frames,
are zero pressure difference, p(r, 4, 0) ¼ p(r, 4, L), such that the flow is
caused solely by the motion of the sphere.
Concerning the boundary conditions along the tube walls, r ¼ R, no-slip
boundary conditions in the fluid frame, v(R, 4, z) ¼ 0, turn into
vðR; 4; zÞ ¼ Ubz in the sphere frame. Full-slip boundary conditions,
vr(R, 4, z) ¼ vrv4(R, 4, z) ¼ vrvz(R, 4, z) ¼ 0, remain the same in both
reference frames. Partial-slip boundary conditions in the fluid frame read,
vr(R, 4, z) ¼ 0 and v4(R, 4, z) þ ‘vrv4(R, 4, z) ¼ vz(R, 4, z) þ
‘vrvz(R, 4, z) ¼ 0, where ‘ is the slip length. In the sphere frame, the right
hand side of the last two conditions is changed from zero to Ubz . The actual
application of the partial-slip conditions requires further consideration due to
software limitations. We use the method presented in Wolff et al. (43): a thin
fluid layer of width e ¼ 0.001R is added around the tube. The additional layer
has a different viscosity, he < h. The boundary conditions are no-slip at the
external radius, R þ e, and full-slip at R. The relation between the slip length
and these parameters is ‘ ¼ e(h/he 1) þ O(e2) (43).
RESULTS
Single sphere in a closed tube
Typical trajectories of fluid particles dragged by a moving
sphere along the central axis (r ¼ 0) of a closed tube
with no-slip boundary conditions are depicted in Fig. 2 a.
At a particular radial position, r ¼ r0, fluid particles
move outward and circle back to their initial position
(Fig. 2 a, inset). Fluid particles located near the center
line (0 % r < r0) are dragged forward, whereas those at
the periphery (r0 < r < R) move backward. The value of
r0/R depends on a/R and L/R.
c
Cytosol Drag in Neurons
We define a net fluid displacement, d(r), as the total
distance traversed by a fluid particle that started at
position r, for given initial and final positions of the moving
sphere, zi and zf. The inset of Fig. 2 a shows three trajec-
tories with positive, zero, and negative net displacements.
The displacement profile corresponding to the trajectories
of Fig. 2 a is shown in Fig. 2 b. The displacement is
zero at r ¼ r0, and for a no-slip tube, it becomes zero again
at r ¼ R.
We now discuss additional general properties of the net
displacement profile. 1), It is independent of U and h.
This can be shown by dimensional arguments and is also
verified numerically. 2), If the initial and final positions of
the sphere, and the initial position of the fluid particle, are
all well separated, zi  z  zf, with jzf  zij [ R, the
displacement profile becomes independent of axial position
d(r) ¼ d(r,4). This results from the fact that the fluid
velocity field decays exponentially with z/R away from the
moving sphere (50). In addition, in the case of azimuthal
symmetry (the sphere moving along the central axis), the
profile depends only on the radial position of the fluid par-
ticle, d(r) ¼ d(r). 3), For the same reason, unless the fluid
particle is arbitrarily close to the center line, the net
displacement is achieved during a finite period of time.
This local response in space and time, resulting from the
confinement of the fluid, allows us to restrict the discussion
hereafter to spheres that move along the entire tube (zi ¼ 0,
zf ¼ L [ R). In this limit, d/R becomes a function of r/R
and a/R only. 4), For sufficiently small values of a/R and a
finite value of r/R, we recover the displacement profile
in an unbounded fluid (53), with d proportional to a,
as shown in Fig. 2 c. For small values of both r/R and
a/R, the displacement profile has the form d ¼ c1a2/r
c2aln(r/R) (53). 5), In a closed tube, due to mass con-
servation, the integral of the displacement profile vanishes,
! d (r, 4) rdrd4 ¼ 0.
The shape of d(r) (Fig. 2 b) may be divided into three
regions. For r(a, the displacement is governed by the
near field—the stage of motion when the sphere and fluid
particle were close together. This behavior is similar to
that in an unbounded fluid, with a displacement proportional
a b
FIGURE 3 (a) Displacement profile for no-slip (black), full-slip (red), and partial-slip tubes with several ‘ values (blue). (b) Trajectory of a fluid particle,
initially located at r‘, for no-slip (black), full-slip (red), and several finite values of slip length, ‘. Parameters: a/R ¼ 0.3, L/R ¼ 30. To see this figure in color,
go online.
2713
to a times a function that diverges as r / 0. For a(r(r0,
the displacement is proportional to R, arising mainly from
the stages outside the near field. Finally, for r0 (r<R, the
(negative) displacement is governed by the opposite flow
due to mass conservation.
We now consider the effect of partial slip at the tube
walls. Fig. 3 a depicts the displacement profiles for several
values of the slip length ‘, compared against the profile of a
no-slip (‘ ¼ 0) and full-slip (‘ / N) tube.
An important observation is that the effect of wall slip on
the displacement profile is minor. In addition, we find a
special radial position, r ¼ r‘, at which the fluid displace-
ment is a constant independent of ‘ (see Fig. 3 a, inset).
Closer to the axis, r < r‘, the net displacement increases
as ‘ is increased, whereas closer to the boundary, r > r‘,
the net displacement becomes more negative with the slip.
We note that the insensitivity to ‘ is restricted to the total
displacement. The detailed trajectories of fluid particles
significantly depend on slip, as demonstrated in Fig. 3 b.
Another way to examine fluid drag is through the drift
volume (57), defined as the volume enclosed between the
initial and final positions of a marked fluid surface—here
taken as the cross section of the tube. In the case of a closed
tube, described here, the drift volume vanishes due to mass
conservation, as explained above. We define the positive and
negative contributions to the drift volume, Vpos and Vneg,
which are the integrals of the displacement profile, d(r),
for 0 % r % r0 and r0 % r % R, respectively, such that
Vpos ¼ jVnegj. Fig. 4 shows the change in these partial
volumes as the slip length, ‘, is varied. Although they
increase monotonically with ‘, the increase is not very
significant, reflecting again the insensitivity of the net fluid
motion to the boundary conditions at the tube wall.
Many spheres in a closed tube
For a system with N simultaneously moving spheres in a
closed tube, we introduce an approximate solution and
verify it as valid if the separation between the spheres is
larger than the tube radius R. Due to hydrodynamic
screening, the fluid trajectories and the displacement profile
Biophysical Journal 106(12) 2710–2719
2714
FIGURE 4 Change in the partial volumes, Vpos ¼ jVnegj, as ‘ is varied.
Parameters: a/R ¼ 0.01, L/R ¼ 6.
can be calculated by superimposing the motions induced by
each sphere separately. Furthermore, for trajectories and
displacements occurring sufficiently far away from the
tube walls, one gets good estimates by considering the
much simpler situation, where each sphere moves along
the symmetry axis of the tube. One subsequently shifts the
resulting fluid motion back to the actual cross-sectional
position of the sphere before superimposing the effects of
all the spheres. This approximation does not satisfy the
boundary conditions at the tube walls, yet the error is found
to be small. For example, the total fluid displacement profile
is calculated according to
X
N
dN ðx; yÞy d1 ðx  xi ; y  yi Þ; (2)
i¼1
where (xi, yi) denotes the cross-sectional position of the ith
sphere, and d1 is the displacement induced by a single
sphere moving along the central axis as obtained, for
example, numerically.
We demonstrate the superposition approximation for 11
spheres arranged as shown in Fig. 5, and Fig. 6 compares
the displacement profiles, d(r), as obtained from the full
numerical solution and from the superposition approxima-
tion in Fig. 5. The approximation error is observable but
small, on the order of a few percent.
FIGURE 5 Geometry of 11 spheres arranged at five different initial
radial positions. (a) Side view. (b) Cross-sectional view. Parameters:
a/R ¼ 0.1, L/R ¼ 12.
Biophysical Journal 106(12) 2710–2719
Mussel et al.
Open-ended tube
In an open-ended tube with zero pressure difference
between the tube ends, the sphere movement generates in
the entire tube a z- and 4-independent global flow, vðrÞ, in
addition to a fast-decaying local flow. The latter screened
contribution is very similar to the flow produced by the
sphere in a closed tube, as described in the previous section.
This is demonstrated in Fig. 7.
Consequently, the superposition approximation for N
spheres in an open-ended tube takes the form
X
N
dN ðx; yÞy ½d1 ðx  xi ; y  yi Þ þ vi ðrÞti ; (3)
i¼1
where d1 is the single-sphere displacement profile discussed
in the previous section, and ti ¼ L/Ui is the total time of
motion of sphere i through the tube. Since vi is proportional
to Ui/L (58), the contributions of the global flows, vi ti , add
to the net displacement a constant, dðrÞ, independent of
Ui and L.
The contribution of the global flow to the net displace-
ment in an open-ended tube, d, makes it always larger
than the net displacement in the corresponding closed
tube. In addition, it makes the displacement sensitive to
the boundary conditions on the tube wall. Both effects can
be seen in Fig. 8. As the slip length is increased, the net
displacement increases because of the reduced friction at
the walls. Its profile flattens with increasing ‘, as the global
flow changes from parabolic to plug-shaped. For full-slip
boundary conditions, the net displacement becomes propor-
tional to the system size, L. Without the effect of the global
flow, the curves shown in Fig. 8 become similar to those in
Fig. 3 a.
DISCUSSION
Consequences for cytoplasmic drag in axons
Transport due to the motion of a single cargo
Let us estimate the mean displacement of a soluble molecule
due to the motion of a single cargo. We assume a uniform
distribution of ~100 microtubules over an axonal cross sec-
tion of ~1 mm2 (46,59). Each soluble molecule is therefore
within a 50 nm distance of a microtubule. We consider the
displacement of the molecule due to the motion of a cargo
on the nearest microtubule only. Including cargoes moving
farther away will only increase the mean displacement,
and the estimate given below, therefore, is a lower bound.
The mean displacement induced by a cargo moving along
the nearest microtubule (i.e., averaged over 0 < r <
50 nm) is proportional to the cargo radius (see Results).
Because of the slow radial decay, the proportionality factor
is large (of order 10), as demonstrated in Fig. 9. The conse-
quence is that a single cargo induces a ballistic motion of the
Cytosol Drag in Neurons
fluid near the microtubule to a distance an order of magni-
tude larger than the cargo itself, which is of order 1 mm.
Since the cargo moves at a typical velocity of 1 mm/s, this
motion should last for a few seconds.
The rate of cargoes traveling along an axon was
measured to be ~1–100 cargoes/min in axons with a cross
section of 1 mm2 (60,61). This is a lower-bound estima-
tion, since not all cargoes are visible in such experiments.
We therefore consider a cargo rate of the order of
100 cargoes/min. Assuming that the cargoes are uniformly
distributed among the microtubules, the rate of cargoes on
a single microtubule, g, is ~1 cargo/min. Thus, on average,
a soluble molecule is dragged ballistically for a few
seconds every minute. In addition to the significant ballis-
0.05
1.5
0
1
-0.05
0.4 0.5 0.6
0.5
ρ0/R
-0.50 0.2 0.4 0.6 0.8
FIGURE 7 Fluid trajectories due to the motion of a single sphere in
no-slip closed and open-ended tubes. Red trajectories are for an open-ended
tube after subtraction of the global (Poiseuille) flow, and black trajectories
are for a closed tube. (Inset) Enlargement of trajectories near r0.
Parameters: a/R ¼ 0.1, L/R ¼ 10. To see this figure in color, go online.
2715
FIGURE 6 (a) Displacement profile of the
system described in Fig. 5 as obtained numerically.
(b) Approximate displacement profile as obtained
from superposition. (c) Difference between the
two displacement profiles. To see this figure in
color, go online.
tic motion, the soluble molecule is expected to undergo
smaller drags in both directions due to cargoes on more
distant microtubules.
In summary, our lower-bound estimate yields an occa-
sional micron-scale ballistic motion (stop-and-go motion)
of a soluble molecule, lasting a few seconds. These
results are consistent with those from experiments by
Roy et al. (31,34). (Note that in those experiments, the
analysis was deliberately restricted to proteins moving
along large distances of >10 mm.) For illustration,
Fig. 10 shows two such intermittent trajectories of a
soluble molecule due to cargo motion that bear qualita-
tive resemblance to trajectories reported in the Roy
et al. studies (31,34).
FIGURE 8 Displacement profiles induced by a single sphere in an open-
ended tube for several slip lengths. The case of a no-slip closed tube is
shown for reference. Parameters: a/R ¼ 0.1, L/R ¼ 10.
Biophysical Journal 106(12) 2710–2719
2716 Mussel et al.

complete unidirectional transport (e.g. for Macropinosomes

0 < ρ/R ≤ 0.05 or mitochondria in specific regions) (64). As an example, a
4
0 < ρ/R ≤ 0.1
10% bias would give a mean fluid displacement of
0.1hdi/min, which is of the order of 0.1–1 mm/day for a
3
closed tube. This is a conservative estimate; describing the
axon as an open-ended tube with partial-slip boundary
2 conditions would yield a larger net flow (note the effect in
Fig. 8). These values are not inconsistent with the
1 experimentally observed overall rate of SC particles: 0.1–
1 mm/day for SCa and 1–10 mm/day for SCb (1).
0
0 0.05 0.1 0.15 0.2
Further implications

FIGURE 9 Mean fluid displacement within a given distance from the
microtubule (rmax/R ¼ 0.05 or 0.1) as a function of a/R in a closed tube.
Parameter: L/R ¼ 15.
Overall transport
We now turn to the overall transport of soluble molecules
due to the motion of many cargoes. This transport is
crucially dependent on the existence, or absence, of bias
in the direction of cargo motion.
Let us begin by assuming no such bias. In such a case,
fluid particles are transported randomly in both directions.
Over a very long time ([g1, i.e., many minutes), their
motion will become diffusive. The effective diffusion coef-
ficient is Deff ~ ghdi2 ~ 1010  109 cm2/s. This is much
smaller than the diffusion coefficient of water molecules in-
side cells (106  105 cm2/s (62)), and still smaller than
that of proteins (~108 cm2/s (63)) as measured in nerve
cells. Thus, in the absence of bias, the cargo-induced motion
considered here does not contribute appreciably to the over-
all transport over very long times. However, as discussed in
the preceding section, it can occasionally make molecules
move ballistically over micron-scale distances—a type of
transport impossible to achieve through thermal diffusion.
On the other hand, a bias in the direction of cargo motion
along the microtubules will create a net drift resulting in the
accumulation of net displacement in the direction of the
bias. Experimentally, a previous report indicates significant
biases ranging from 10–30% for some cargoes, and up to
In this article, we propose that drag induced by active cargo
motion may be a plausible mechanism contributing to the
motion of SC proteins. The implications for axonal transport
are discussed above. The essential conclusion is that fluid
particles and soluble molecules should be dragged in a
stop-and-go bidirectional motion along micron-scale dis-
tances during times on the order of seconds. This conclusion
is in line with the known experimental facts concerning SC
transport. Although additional mechanisms influencing the
dynamics of SC proteins are not ruled out, the mechanism
proposed here relies on simple physical effects without the
need to add extra elements such as binding of the molecules
to motor proteins (35).
Our analysis is based on two main assumptions. The first
is that transported molecules are assumed to be well dis-
solved in the medium and therefore carried along with it
as it flows. The theory does not hold for molecules that do
not conform to this assumption, e.g., proteins that bind to
intracellular structures.
The second major assumption is that the axonal medium
is fluid, i.e., it should flow in response to the active motion of
the cargo. Such flows were observed in response to the
forced motion of magnetic particles (65). Evidently, the
actual axoplasm is crowded, viscoelastic, and anisotropic
(65). We have focused in this work on the simple question
of whether the passive drag due to cargo motion is at all rele-
vant or negligible as regards the SC of axonal transport.
Within this modest goal, the orders of magnitude presented
above should be valid for actual axons, based on the

a 0.35 b 0.7
0.6
0.3 #2
0.5
FIGURE 10 Two demonstrative trajectories of
0.25
#3 0.4 fluid particles along the axonal direction, as
0.3 induced by the motions of three cargoes. Parame-
z [μm]

0.2
z [μm]

#1 #3
0.2 ters: R ¼ 1 mm, a/R ¼ 0.1; cargo coordinates:
0.15
0.1 (r1, 41) ¼ (0.5R, 0), (r2, 42) ¼ (0.1R, p) moving
0.1 0
#1 in the opposite direction, and (r3, 43) ¼ (0.4R,
-0.1 p); fluid particle coordinates: (ra, 4a) ¼ (0.345R,
0.05 #2
-0.2 0), (rb, 4b) ¼ (0.375R, p). To see this figure in
0 -0.3 color, go online.
0 2 4 6 8 10 12 0 2 4 6 8 10 12
time [s] time [s]

Biophysical Journal 106(12) 2710–2719

Cytosol Drag in Neurons
following arguments. 1), The complex forces exerted on
the cargoes by molecular motors, and their resulting veloc-
ities—the well-documented fast component of axonal trans-
port—are not addressed theoretically but are rather treated
as external biological constraints imposed on our calcula-
tions of the drag of the much smaller dissolved proteins.
2), It is well established that confinement in a long tube
must screen out hydrodynamic interactions over long dis-
tances—an effect that is directly related to the lack of
momentum conservation in the confined fluid and remains
valid regardless of the complexity of the fluid. 3), The envi-
ronmental response to an object moving through the axon
depends on size. For objects of molecular size, such as pro-
teins, the medium’s response was found to be close to that of
a simple solvent (66). 4), Concerning the long-distance flow
induced by cargo motion, viscoelastic effects should set in
at sufficiently large length- and time-scales. The only mea-
surement of axoplasm viscoelasticity of which we are aware
(65) gave comparable values for the storage and loss moduli
over timescales of order 103 s. Measurements in the cyto-
plasm of other cells (macrophages) (67) yielded a storage
modulus of the same order as, or one order of magnitude
larger than, the loss modulus for timescales of order 10 s.
Related materials such as F-Actin networks are known to
have a storage modulus that increases with time, where
the crossover from viscous to elastic behavior takes place
at ~1–10 s (68). Thus, the relevant timescale of cargo mo-
tion, ~1 s, may be around that crossover; we expect visco-
elastic effects to be present but not very large. Clearly,
more accurate experimental data for the viscoelasticity
of the axoplasm is required. We further note that in the
noninertial (zero Reynolds number) limit considered here
for given cargo velocities, the results for the displace-
ments (rather than stresses) of the dragged molecules are
independent of the viscosity. Qualitatively, in a viscoelastic
medium, they should be similarly insensitive to the fre-
quency-dependent moduli.
Another issue, addressed in detail in the Results section,
is the effect of boundary conditions along the axonal mem-
brane, namely, whether or not there is significant slip at the
boundary. We find that slip may be either unimportant or
important for axonal transport, depending on whether the
axon should be described as effectively closed or open-
ended, respectively (compare Figs. 3 a and 8). This issue,
in turn, depends on the amount of water exchange across
the axonal membrane. Thus, the issues of boundary slip
and membrane permeability are intertwined and should be
clarified in future experiments to obtain a more quantitative
description of the transport studied here.
An important implication of this study relates to the
interpretation of nonlocal measurements of intracellular
diffusion of various proteins, such as by FRAP or diffu-
sion-weighted NMR (69). It should be taken into account
that intracellular diffusion coefficients extracted from such
measurements in vital cells may be overestimated due to
2717
the contribution of active mechanisms that may induce
microstreaming. Experiments that involve fixation of cells
or blockage of various possible active processes (35) can
provide the needed discrimination between diffusion and
microstreaming. Only with additional experiments of this
sort will we be able to quantify the relative contribution of
thermal diffusion and active mechanisms in defining the
SC and intracellular displacement of soluble proteins.
The implications of water displacement in neurons extend
beyond the basic understanding of transport and axonal
physiology. Diffusion tensor imaging (DTI) uses water
displacement in the brain as a unique tool for diagnosis
and research (70). Water displacement in neuronal projec-
tions is anisotropic (with preferred displacement in the lon-
gitudinal direction), and significantly drops during events of
metabolic or oxidative stress, as occurs, for example,
in cerebral artery occlusion (71). DTI measures water
displacement over typical timescales of 1–100 ms. Our
calculation implies that, contrary to a previous suggestion
(72), within these timescales, the displacement of water
molecules, caused by drag due to transported cargoes, is
too minor to be detected by DTI. On a more general level,
it should be noted that some of the displacement of water
molecules, as well as the drop in displacement detected after
tissue insults, may be due to blockage of intracellular micro-
streaming. The existence of such active microstreaming, as
well as its biophysical basis, should be further investigated.
Transport mechanisms similar to those proposed here
might be relevant to the mixing and distribution of solutes
in other scenarios. A particular example is that of cyto-
plasmic streaming in plant cells (56,73–75). Organelle-
induced flow is observed in many plant cells in which
different flow patterns with variable velocity distributions
have been identified. Nonetheless, organelle-induced flow
is expected to be most prominent in cells of anisotropic
cytoskeleton, where there are displacements over long
distances. These conditions are met in neurons.
We thank S. Bhattacharya, A. Leshansky and E. Perlson for helpful
discussions, and A. Liberman for his assistance.
H.D. acknowledges support from the Israel Science Foundation (grant no.
8/10). U.N. acknowledges support from a European Union Marie Curie
Institutional Research grant (MMDTIAN) and an Israel Science Foundation
grant (1156/12).
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