Text of the sticky note comment/annotations in the pdf.

(in case you cannot open the notes)

(page 1 note)This simple, descriptive paper is a landmark because it documents a type of cell
motility (rearward movement of particles on the cell surface) which is still under active
investigation today. Please
Note the two possible mechanisms discussed (in discussion) because we will later show
evidence that their preferred mechanism is wrong, and the other, less likely one turns out to be
correct.
(Comment re material and methods) For all the papers we will read, the materials and methods
section is
critical.
Queries above Fig. 1.
What is a fibroblast and what is its function in vivo
In these phase contrast LM images, consider which regions of the cell imaged has a) lowest
refractive index, b) intermediate refractive index and c) highest refractive index.
All the figures in this paper will be discussed. Ask what is the question asked, what does the
data show, and when relevant, what control experiments were performed.
(Query re. Fig. 4: Why is this a critical/key result?
Enzyme treatments discussed at end of results section) Look up these enzymes and discuss
why they were chosen; although there is no clear answer, think about why collagenase might
increase frequency of particle attachment/transport (note not the speed--just frequency)
pg 395: this is the key section where they discuss possible mechanisms for rearward particle
movement

Experimental

THE LOCOMOTION
III.

Cell Research 62 (1970) 389-398

OF FIBROBLASTS

IN CULTURE

Movements of Particles on the Dorsal Surface

of the Leading Lamella

M. ABERCROMBIE,

JOAN E. M. HEAYSMAN

and SUSAN M. PEGRUM

Department of Zoology, University College, London, London WCI, UK

SUMMARY
A fibroblast, when moving on a plane substratum and colliding with a small particle adherent
to the substratum, commonly picks up the particle and transports it backwards. Such particles
while moving back are attached to the dorsal surface of the cell, i.e. to the side exposed to the
medium. On-the average, in about 8 min they travel back aboui 14 pm with reference to the
front edge of the cell and about 10 ,um with reference to the substratum. showing little fluctuation in s-Fed or direction. They thin go into random oscillation or mo;e sideways. The backward speed of particles is approximately equal to that of ruffles in a similar situation, but it
is significantly slower than that of large pinocytotic vacuoles. It is proposed that the lamellipodia at the front end of a cell result from outbursts of assembly of new surface there, producing folding, the excess surface steadily flowing backwards on the dorsal side of the cell, where
it is unimpeded by adhesions to the substratum, carrying the particles with it.

At the front end of the leading lamella of a

fibroblast during its locomotion on a plane
substratum there occur thin mobile lamelliform projections which we have termed
lamellipodia
[2]. They show consistent patterns of protrusion, withdrawal and bending
[l, 2, 71 which give rise to fluctuations in the
position of the anterior edge of the cell and
to the phenomenon of ruffling. When such
a lamellipodium
forming the front end of a
fibroblast collides with a small particle loosely adhering to the substratum on which
the fibroblast is moving, the particle may be
detached and moved on to the cells dorsal
surface (i.e. the surface exposed to the liquid
medium). The present paper analyses the
movement of such particles. The analysis
leads us to propose that lamellipodia
are
regions of rapid assembly of new surface.

MATERIAL

AND METHODS

A drop of culture medium [l] containing a very

little carbon (Gurr India Ink for injection purposes,
diluted to 1O-6 and the large particles then removed
by centrifugation),
or a very little finely ground
molybdenum disulphide [5], was spread on each of
a series of coverslips. The preparation was stood for
several hours with the drop above the coverslip.
Then the medium with any particles that had not
settled was removed, and fresh medium was added,
with a fragment of heart from a 7-9 day old chick
embryo. The culture was subsequently incubated as
a hanging drop, so that only adherent particles remained on the coverslip. The carbon particles averaged 0.7 pm in diameter (range 0.5 to l.O), the molybdenum disulphide particles averaged 1.0 pm (range
0.5 to 1.6). Both kinds of particles behaved alike,
and they are not distinguished in our analysis;
debris, consisting probably of fragments of cells [16],
also behaved in the same way as the artificial particles. After overnight incubation, when fibroblast-like
cells had emerged from the explant, observations were
made of collisions between cells and particles. Some
collisions were filmed, with time-lapse methods
previously described [l]. In these films the movements
of three points on the front edge of each cell were
analysed by projecting the filmed cell on to three
Exptl

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390

M. Abercrombie et al.

Fig. I. Stills at 75 set intervals from a film. In frame 1 contact between cell and particle (arrciw) has not been
made. In frame 2 contact is made but the particle has not moved. In subsequent frames the right-hand arrow
indicates the original position of the particle; the left-hand arrow the present position. In the later frames the
particle moves out of focus as it reaches the thicker posterior part of the leading lamella. This particle ultimately reached the side of the cell, where it could be seen in profile. x 850.

lines lying parallel to the direction of cell movement

and at a distance apart representing 6.25 pm [l]. One
of the reference lines was made to coincide with the
particle in the frame where collision occurred. The
positions of the cell edge where it cut the lines, and of
the particle, were recorded every half-minute.
Exprl

Cell Res 62

RESULTS
The experimental arrangement consisted of
chick heart fibroblasts moving on a glass
substratum, to which particles were adher-

Movements of particles on fibroblasts.

ent. As a fibroblast moved it collided from

time to time with a particle. Two hundred
such collisions were observed in a qualitative
way. In 86 instances, the particle remained
where it was, the cell evidently moving over
it. In 4, the particle was detached from the
substratum by the collision, and immediately,
since the culture was a hanging drop, fell
off into the medium. In 1, the particle was
pushed forwards. In the remaining 109, very
soon after visible contact between cell and
particle, the particle started to move in relation to both cell and substratum, travelling
backwards while apparently adhering to the
dorsal surface of the cell. An example is
illustrated in fig. 1. Only one of these particles
became detached from the cell and fell into
the medium during the 5 to 10 min that each
was observed.
It is backward movement of these particles
that we propose to analyse in this paper in
relation to the other features of cell movement previously described [l, 21. It is first
necessary to consider whether the cells used
for this analysis were similar in their movement to the unmarked cells previously described. or whether the marking had altered
them or the sampling had been different.
Fourteen particles on twelve cells were filmed.
The average speed of translocation,
with
standard error, of these cells was 0.55kO.12
,um/min; that of the 13 unmarked chick heart
cells previously analysed [l, 21 was 0.60*
0.09; the difference is obviously not significant. The front edge of the marked cells
spent a somewhat higher proportion of the
time stationary (59 * 4 %, as against 50 * 3 %
in the unmarked cells; for the difference t =
1.89 degrees of freedom (d.f.) 25, 0.1 > P>
0.05). As a result, the frequency of the protrusion-withdrawal
fluctuations of the edge
[l] was a little lower in the marked cells
(18.1 k2.5 fluctuations/100 min in the marked
cells against 23.4k2.5 in the unmarked;

III

391

i
2

3
2

4
3

)2

222

1
\2

1Opm

Fig. 2. Pathways of a sample of particles, recorded

at half-minute intervals. The arrow represents the
direction of cell locomotion in all c&es; particle
movement is opposite. The numerals indicate points
where the particle was recorded for the indicated
number of readings, and was therefore stationary.

for the difference t = 1.50, d.f. 25, 0.2> P>

0.1). The average amplitude of the fluctuations did not differ between the marked and
unmarked groups. The only significant difference detected was that the marked cells
had many fewer ruffles than the unmarked.
This difference was due to sampling, not to
the experimental conditions. Cells with few
or no ruffles were at first chosen for recording because on them the particle was more
clearly visible, and because we wished to
eliminate the possibility that the ruffles by
their own backward movement [2] transported the particles. We later added for
comparison a small group of cells in which
ruffling was active.
The movement of each of the particles in
the 14 specimens analysed was followed for
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392

M. Abercrombie

et al.

a somewhat arbitrary time, averaging 8 min,

during which it proceeded fairly directly
backwards; when movement of this kind
ceased analysis was stopped. Specimens of
the tracks of particles, recorded at halfminute intervals, are shown in fig. 2. Ten of
the 14 particles travelled with only momentary deviations directly backwards during the
period of measurement, so that their mean
track was almost exactly parallel to the cells
direction (which did not change during this
time). The other 4 moved diagonally backwards, at between 20 and 30 to the course
of the cell, again with only minor deviations
of direction. Two of these, and one of those
that travelled straight back, changed direction at the end of the period of measurement
through almost a right angle, and proceeded
to move approximately transversely to the
cell. The rest went into roughly random
oscillations, or, in one case, fell off the cell.
During the period of relatively direct backward movement, the particles travelled back
on average about 10 pm with reference to
the substrate, while during the same period
the front edge of the cell advanced on average about 4 pm. When backward movement
ceased the particles were between 6 and 25
,um behind the front edge, where they usually remained, in a region in front of the
nucleus where the leading lamella merges
into the thicker part of the cell body.
The average speed of travel of the 14 particles with reference to the substratum, regardless of direction, was 2.03 ,um/min
(SD = 1.00). The average of the speeds of
the backward component of the particles
movement, that is, the movement diametrically opposite to the direction of movement
of the cell, was 1.69 pm/min (S.D. =0.75)
(2.24, S.D. =0.78, with reference to the front
edge of the cell). The predominance of the
backward direction is obvious: the net backward movement represented 84 % (S.D. =9)
ExprI

Cd

Res 62

of the total distance travelled. The maximum

speed in the backward direction recorded
over a half-minute interval was 6.8 ,um/min.
The speed of the particle was clearly correlated with the amount of ruffling of the cell.
In the relatively unruffled group of cells (9
specimens of which 5 showed no ruffle on
the three reference lines during the movement
of the particle, the others having up to 13 %
of readings manifesting a ruffle) the mean
particle backward speed was 1.32 pm/min
(S.D. =0.58). In the group of 5 cells chosen
to examine the effect of ruffling (which was
manifested in 18 to 90 % of readings) the mean
particle speed was 2.34,um/min (S.D. =0.59).
The difference between the two groups is significant (t=3.13; degrees of freedom (d.f.) =
12; PCO.01).
In the half-minute immediately following
apparent contact with the cell, the particle
was stationary in 5 cases, and moved slightly
forwards in 1; in the other 8 it had already
begun its backwards movement. During this
same half-minute the part of the front edge
of the cell that makes contact with the particle
was usually stationary. In only 2 of the 14
cases did the edge continue to move forwards immediately after the contact, while
in 1 it reversed. The rather high proportion
that came substantially to a standstill suggests that the apparent contact was a reality.
Where the particle immediately
began its
backward travel, the edge simultaneously
moved forward in 2 instances, backward in 1
and was stationary in 5.
After the frequently occurring initial hesitation, the particles showed no significant
change in their speeds of movement, either
gross speeds or net backward speeds, until
near the end of their travel when irregular
movement starts. The edge of the cell, after
its corresponding initial hesitation, resumed
its normal fluctuations (fig. 3). During the
period of direct backward movement of the

Movements

of particles on fibroblasts.

III

393

still not significant. But the correlation is

significant when confined to the ruffled group
during this initial period (r = + 0.397; d.f. =
30; 0.05>P>O.O2).

10.0

Fig. 3. Abscissa: min; ordinate: ,um before and behind point of collision.
Movement of the cell edge in relation to movement of the particle in a typical instance. The upper
line ( x - x - x ) represents the position of the front
edge of the cell, undergoing its usual fluctuations.
The lower line (O-O-O)
represents the more regular movement of the particle.

particles, 23 % of the half-minute intervals

showed the particle stationary; it was rarely
stationary for more than one interval in succession. This is a much lower proportion of
time stationary than is manifested by the
front end of the cell (59 % of intervals), but
the recording of movement of the particle is a
good deal more sensitive than the recording
of movement of the cell edge. The much
greater consistency of backward movement
of the particle is best demonstrated by the
forward movements, which take up 24% of
the time in the case of the cell edge, but only
3 % in the case of the particles. The pattern
of movement is indeed totally different, and
highly significantly so when tested by chisquare. The correlation
between particle
backward speed and the simultaneous backward speed of the cell edge directly in front
is quite non-significant (r = + 0.023; d.f. =
229). If the correlation is confined to the
first 4 min of particle movement (after the
initial half-minute), when the particle is still
near the edge, r = +O. 127, d.f. = 88, which is

The evidence that during their backward

movement the particles were on the dorsal
surface of the cell is as follows. (1) Towards
the end of its direct movement, when particles were situated where the cell was thick,
their plane of focus with a phase contrast
objective of N.A. 1.4 indicated a situation
on the dorsal surface. (2) In two of the instances analysed in detail, the particle was carried to the side of the cell, after its backward
movement, and it there appeared in profile
projecting from the cell margin. (3) A particle was observed by Ingram [7] during his
study of fibroblasts in side view, and was
clearly on the dorsal surface of the cell. (4)
Particles have been found in electronmicrographs of sections of cells known to be transporting particles backwards in the usual
way; they were on the dorsal surface (fig. 4),
separated by several hundred Angstroms
from the unit membrane.
How does the particle movement compare
with other backward movements recorded in
this region of the cell? The withdrawal movement of the front edge previously analysed
[l] is clearly a good deal faster.- In the cells
bearing the particles the mean speed of each
of 69 withdrawal phases of the protrusionwithdrawal fluctuation was measured; the
mean of these was 4.9 pm/min (S.D.=2.0).
The backward movement of ruffles when they
lie behind the front edge of the cell, and
therefore in the region in which the particles
are moving at least at the beginning of their
travel, was previously [l] found to average
2.8 pm/min (N=27; S.D. =3.1). This is similar to the mean of 2.3 pm/min recorded
for particles on cells showing ruffling; the
maximum speed was 7.5 pm/min for ruffles,
6.8 pm/min for particles. Too few data are
Exptl

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0.5

,um.

available to estimate the movement of ruffles

while behind the edge in the cells actually
used for particle analysis, but the general
backward movement of ruffles on these cells
averages 3.2 pm/min
(N=17;
S.D.=3.7).
While mean particle speed does not differ
significantly from these mean ruffle speeds,
the variances do differ significantly; the standard deviation of particle speed is only 0.6.
Another conspicuous backward movement in
some cells is that of large pinocytotic vacuoles, which are taken into the cell amongst the
ruffles near the front edge, and move back
a short distance in the leading lamella before disappearing from view. In the cells
bearing the particles, 23 such vesicles were
measured. Excluding one exceptionally fast
vesicle (which moved 17 pm in 1.5 min),
the average distance moved with reference to
substratum was 5.8 pm (S.D.=2.3) at an
average speed of 3.4 ,um/min (S.D. =0.9).
This is significantly faster than the movement
of particles on ruffle-bearing cells (t =2.52;
d.f. = 26; 0.02 > P > 0.01). A generalised
backward movement is sometimes visible in
time-lapse films of the anterior end of fibroblasts (and other kinds of cells), unmarked
by particles. What one sees is a series of
indefinite shadows chasing each other steadily backwards from the edge in the region
where particles move; but it proved impossible to put any reliable estimates of speed
on the movement, though it is not widely
Exptl

CeN Res 62

different from that of the particles. Finally,

we have noticed that on the strands of cytoplasm radiating from a cell that has rounded
up during mitosis, swellings can often, but
not always, be seen travelling centripetally.
Measurements on 12 of these showed a mean
speed of 17 pm/min, a different order of
magnitude from that of the particles.
We briefly mention here that we have
made a number of attempts to interfere with
the adhesion of the particles to the cells, and
hence with their transport, by exposing the
cells to enzymes in the liquid medium while
they were moving amongst particles. Taking
as an index the frequency with which collision with a particle was followed by its backward transport, we found hyaluronidase
(Schwarz BioResearch, 200 USP units/ml)
and phospholipase C [8] (Koch-Light, 0.2
units/ml) without effect. Collagenase (Schwarz
BioResearch, 14 units/ml) significantly increased the frequency of particle transport.
DISCUSSION
We have presented evidence that when the
lamellipodium
at the front edge of a moving fibroblast collides with a particle that is
sufficiently loosely adherent to the substratum, the particle is transferred to the dorsal
surface of the cell and there moves for some
distance steadily backwards, not only in relation to the cell, which continues to move

Movements of particles on fibroblasts.

forwards, but in relation to the substratum.

It may be noted that a backward movement
appearing as a steady flow over a distance
similar to that travelled by the particles, can
sometimes be seen in time-lapse films at
the front end of a cell without any artificial
marking. We have not, however, been able
to measure the speed of this unmarked movement, so we cannot safely equate it with the
particle movement.
Does the particle movement represent a
bodily movement of the cell surface, or is the
particle somehow propelled independently?
The particles are of course strongly adherent
to the cell surface. These were hanging drop
preparations, and yet particles as dense as
molybdenum sulphide almost never fell into
the medium while moving backwards. A
sliding adhesion is however conceivable. If
the particle moves independently of transport
of surface material, it is most likely to be by
the agency of backward-moving waves of deformation in and vertical to the cell surface.
Could the ruffles represent such waves? The
waves would need to move backwards a good
deal faster than the particles; and since the
displacement of the particle would occur in
a series of backwards and forwards oscillations and such oscillations are not observed,
the frequency of the waves must be high.
The visible ruffles themselves could not be
the waves involved, since particle movement
occurs without them. But neither are the
ruffles momentary exaggerations of invisibly
small waves that cause the movement: their
speed of propagation is hardly different from
that of the particles. If the waves exist, they
have so far given no other sign.
We provisionally conclude that the most
likely hypothesis is that the cell surface to
which the particles stick moves bodily along
with the particles. It is evident that such surface movement does not merely reflect the
protrusion-withdrawal
fluctuation
of the

III

395

front edge of the cell: forward movements

of the particle are extremely rare. It seems
that, granted surface flow, new surface must
appear in front of the particle; and if this is
so one is led to the tentative conclusion that
new assembly of surface is continually occurring in front of the region where the ventral
side of the cell adheres to the substratum,
that is, in the immediate neighbourhood of
the front edge of the cell, where the lamellipodia protrude.
We therefore propose the general hypothesis that lamellipodia,
the thin, veil-like,
transitory, shifting projections of cells, wherever they appear, are regions of assembly of
excess surface material, both unit membrane
and coat, which is occurring sufficiently
fast to produce folding of the surface; the
excess surface so formed flowing away, as a
rather coherent sheet, from the place of its
assembly to an unknown sink. According to
this hypothesis the protrusion-withdrawal
fluctuations of the anterior, approximately
horizontal, lamellipodium
represent random
and localised fluctuations in the rate of surface assembly; perhaps also to some extent
in the rate of flow away, though the particle
movement suggests this is rather constant,
continuing often during protrusion of the
edge, occurring in fact with a speed uncorrelated with edge movement (when ruffles are
absent). The flow away is asymmetrical, i.e.
it predominantly involves the dorsal surface,
because the adhesions to the substratum,
which lie behind the lamellipodium,
will tend
to block it on the ventral side. The flow evidently occurs at least from the extreme tip
of the horizontal lamellipodium,
since a
particle touched by the tip is moved back;
we assume that the flow, at least during protrusion, is probably forwards on the ventral
side of the lamellipodium.
If protrusion of the front edge is due to
an excess of assembly over flow away, and
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M. Abercrombie

et al.

its withdrawal is due to the converse, the

rate of flow must at least be sufficient to carry
away the amount of surface material that
moves backwards during a withdrawal. For
the period of observation of each particle,
an estimate of the amount of surface that has
to be moved back was obtained from the
total distance of the withdrawal movement
of the cell edge (at a point directly in front
of the position of the particle), doubled because the lamellipodium
is a fold and it is
assumed there is no flow away on the ventral
surface. The resultant prediction of minimum
surface backward movement has a mean of
1.36 pm/min, which is comfortably less than
the mean particle speed (1.69 pm/min);
so
that the necessary amount of surface can be
transported back at the particle speed, provided the flow continues during both protrusion and withdrawal. Rather striking is
the correlation between the predicted minimum surface flow rate and the mean particle
speed in each of the 14 observations. The correlation coefficient is 1-0.75, which is highly
significant (P < 0.01).
We can tentatively interpret ruffling, the
bending dorsally of the terminal horizontal
lamellipodium
[2, 71, on the same basis. Because of the asymmetry of the flow away,
we can assume that surface pressure is
higher on the undersurface of the terminal
lamellipodium
than on its upper surface.
Consequently the lamellipodium
will tend to
curl upwards; and if the pressure gradient
becomes sufficiently steep a ruffle may form.
It is perhaps because of this curling up that
the terminal lamellipodium
does not readily
form an adhesion to the substrate near its
tip [l]; it may in fact also frequently miss
particles that lie in the cells path, allowing
the cell to override them. Curling up may be
exacerbated too by excess feeding of new
surface into the lower as compared to the
upper aspect of the terminal lamellipodium.
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Cdl Res 62

This might happen because the initial curling

up exposes the lower aspect more to impinging surface material moving forwards in the
cytoplasm, which would make the curling
up self-reinforcing, and thus account for the
extreme to which the curling so regularly
goes, turning the horizontal lamellipodium
through 90. We suggest such asymmetrical
assembly in the later stages of curling up
because of the particularly frequent occurrence of a vertical ruffle at the end of a withdrawal phase, just as a new protrusion starts
[2]. Vigour of protrusion is correlated with
amount of ruffling [2], and so is the speed
of particle movement, which is consistent
with the suggestion that rapid surface assembly is concerned in ruffle production.
The new assembly of surface, on this hypothesis, finally becomes concentrated at the
base of the now vertical lamellipodium,
and
a new horizontal lamellipodium
is formed.
The vertical lamellipodium
is then either
flattened by the backward flow (perhaps accompanied by some disassembly of surface),
or is swept bodily back by it before flattening. The backward speed of ruffles when they
are behind the front edge is satisfactorily
similar to that of particles on a ruffled cell;
its higher variance may come from wavering
out of the vertical, or from vagarise of continued feeding of new surface into the base
of the ruffle.
It should be added that the curling up and
other movements of lamellipodia
may be
assisted by some rigidity from an internal
skeleton, as in some filopodial extensions
(microspikes [12, 131); though microtubules
have not yet been identified in lamellipodia.
In chick heart fibroblasts, cultured on a
plane surface, filopodia are uncommon [ll],
though they occur. They too may involve
surface assembly.
It would be tempting to include the backward movement of pinocytotic vacuoles in

Movements of particles on fibroblasts.

the general hypothesis that we propose, by

suggesting that they move through retaining some connection with the moving dorsal surface. Their significantly faster mean
speed, however, imposes caution.
A movement analogous to that of our particles was recorded by Marcus [lo]. HeLa
and chick cells, after infection with Newcastle disease virus so that they acquire a
haemagglutinin,
first show adhesion to red
cells at their edges; and recovery from the
abolition of adhesiveness to red cells produced by specific antiserum first occurs in
the same situation, These results are consistent with the appearance of new cell surface
in positions that may well correspond to
lamellipodia.
Marcus was further able to
show that red cells adhering to the newly
recovered edge subsequently moved towards
the nucleus on the dorsal surface of the cell.
The hypothesis we have proposed is that the
full thickness of the surface, unit membrane
and coat at least, is involved in the assembly
and disassembly. This follows from the fact
that the full thickness is folded, and from the
suggestion that the ruffles are sometimes carried bodily backwards by the flow. Equally
the full thickness of the surface must then be
lost somewhere. The loss is not necessarily
sharply localised where the particles cease
their consistent backward movement; there
may be earlier loss not reflected in particle
velocities because the narrowing of the cell
behind the leading edge reduces the area of
moving surface (by perhaps 50 %, at a rough
estimate). How the loss is accomplished
must be left undiscussed. The dynamics
proposed are however probably compatible
with turn-over studies of the plasma membrane of fibroblasts, given the possibility of
a recirculation of surface components [15].
Measurements by planimetry of the approximate total area of the cell surface (dorsal
plus ventral) and of the region involved in

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397

particle movement show the latter to be

about 20 % of the former (mean of 10 measurements). According to our interpretation,
this proportion needs to be replaced in less
than 10 min; the rest of the cell surface may
of course be highly stable [9], as long as it
does not form lamellipodia.
This hypothesis of the dynamics of lamellipodia can, we believe, explain much of the
data of the present and preceding papers
[l, 21. It would be premature to attempt this
in detail. We should, however, mention two
points where the hypothesis, even if substantially correct, possibly or probably requires supplementing.
(1) The withdrawal phases of the fluctuation of the front edge, and the ruffling that
accompanies them, may not be wholly explicable in terms of surface flow. Ingram [7]
has suggested that contraction within the
cytoplasm is involved, and particularly in
view of the role that such contraction seems
likely to play in cell locomotion, we should
not wish to exlude this possibility.
(2) We have not adequately accounted for
the location and shape of lamellipodia. There
appear at present to be two possibilities, not
mutually exclusive. Perhaps the affinity of
particular parts of the existing surface for
new surface material determines where surface assembly shall occur; and these particular parts may themselves be determined
by the position of adhesions to the substratum (and, in the case of phagocytic cells, to
the object phagocytosed [4]). A second possibility is that the cytoplasm contains a
mechanism for driving new surface material,
and perhaps cytoplasmic material, in particular directions, and this determines where
surface assembly shall occur. We believe
that the recent observations of Vasiliev and
colleagues [14], suggesting a role of microtubules in establishing where lamellipodia form,
may be an important clue to this problem.
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hf. Abercrombie

et al.

The formation and withdrawal of surface

has been proposed as the central feature of
the mechanism of cell locomotion [3]. It will
be apparent that if a source of new surface
is situated in front of an area of adhesion to
the substratum, and a sink behind it; and if
the new surface forms new adhesions, and
the sink removes the surface that made the
old adhesions; then the rest of the cell can in
principle be entrained in a forward movement. We have presented evidence that new
surface is formed at the front end of a moving cell. This may be a phenomenon quite independent of locomotion. But since the position of lamellipodia is so strongly correlated
with the direction of cell movement, and the
vigour of lamellipodial
protrusion and withdrawal is correlated with the speed of movement of the cell [l], it seems reasonable to
suggest that the new surface at the front end,
by forming new adhesions to the substratum,
accounts for the forward spreading of the
fibroblast, and in general for its ability to
spread on a substratum. It is evident that it
spreads rather inefficiently in the circumstances that we have investigated; new adhesions must usually form well behind the
tip of the protrusion to allow the subsequent
relatively free withdrawal. But though we
suggest that ground is gained by the leading

Exptl

Cell Res 62

end of the cell in this way, it seems to us probable that the contractile system of the cell,
rather than the removal of the surface that
has formed the adhesions, plays the major
part in drawing the rest of the cell forwards
16 7, 171.
REFERENCES
1. Abercrombie, M, Heaysman, J E M & Pegrum,
S M, Exptl cell res 59 (1970) 393.
2. - Ibid 60 (1970) 437.
3. Bell, L G E, J theoret biol 1 (1960) 104.
4. Brewer, D B, J path01 bacterial 86 (1963) 299.
5. Clarris, B J & Fraser, J R E, Exptl cell res 49
(1968) 181.
6. Gustafson, T & Wolpert, L, Biol rev 42 (1967)
442.
7. Ingram, V M, Nature 222 (1969) 641.
8. Lesseps, R J, J cell biol 34 (1967) 173.
9. Lucy, J A, J theoret bio17 (1964) 360.
10. Marcus, P I, Cold Spring Harbor symp quant
biol 27 (1962) 351.
11. Overman, J R & Eiring, A G, Proc sot exptl biol
med 107 (1961) 812.
12. Taylor, A C, J cell bio128 (1966) 155.
13. Taylor, A C & Robbins, E, Dev biol 7 (1963)
660.
14. Vasiliev, J M, Gelfand, I M, Domnina, L V,
Ivanova, 0 Y, Komm, S G & Olshevskaja, L V.
J embryo1 exptl morphol. In press.
15. Warren, L & Glick, M C, J cell biol 37 (1968)
729.
16. Weiss, L & Lachmann, P J, Exptl cell res 36
(1964) 86.
17. Wolpert, L, Symp sot gen microbial 15 (1965)
270.
Received April 9, 1970