Actin filaments and their interactions with cell surface molecules have key roles in tissue cell behaviour. Axonal pathfinding during embryogenesis, an especially complex cell behaviour, is based on the migration of nerve growth cones. We have used fluorescence immunocytochemistry to examine the distribution in growth cones, their filopodia and lamellipodia of several actin-associated proteins and nerve cell adhesion molecules. The leading margins of chick dorsal root ganglion nerve growth cones and their protrusions stain strongly for f-actin, filamin, alpha-actinin, myosin, tropomyosin, talin and vinculin. MAP2 is absent from DRG growth cones, and staining for spectrin fodrin extends into growth cones, but not along filopodia. Thus, organization of the leading margins of growth cones may strongly resemble the leading lamella of migrating fibroblasts. The adhesion-mediating molecules integrin, L1, N-CAM and A-CAM are all found on DRG neurites and growth cones. However, filopodia stain relatively more strongly for integrin and L1 than for A-CAM or N-CAM. In fact, the 180 X 10(3) Mr form of N-CAM may be absent from most of the length of filopodia. DRG neurones cultured in cytochalasin B display differences in immunofluorescence staining which further emphasize that these adhesion molecules interact differentially with the actin filament system of migrating growth cones. Several models for neuronal morphogenesis emphasize the importance of regulation of the expression of adhesion molecules. Our results support hypotheses that cellular distribution and transmembrane interactions are key elements in the functions of these adhesion molecules during axonal pathfinding.

BAMBURG
J. R.
,
BRAY
D.
(
1987
).
Distribution and cellular localization of actin-depolymerizing factor.
J. Cell Biol.
105
,
2817
2825
BENTLEY
D.
,
CAUDY
M.
(
1983
).
Pioneer axons lose directed growth after selective killing ofguidepost cells.
Nature, Lond.
304
,
62
65
BENTLEY
D.
,
KESHISHIAN
H.
(
1982
).
Pathfinding byperipheral pioneer neurons ingrasshoppers.
Science
218
,
1082
1088
BERNSTEIN
B.W.
,
BAMBURG
J. R.
(
1982
).
Tropomyosin binding to f-actin protects the f-actin from disassembly bybrain actin depolymerizing factor (ADF).
CellMotility
2
,
1
8
BINDER
L. I.
,
FRANKFURTER
A.
,
KIM
H.
,
CACERES
A.
,
PAYNE
M. R.
,
REHBUN
L.L.
(
1984
).
MAP2 heterogeneity duringrat brain development.
Proc. natn. Acad. Sci. U.S.A.
81
,
5613
5617
BIXBY
J. L.
,
PRATT
R. S.
,
LILIEN
J.
,
REICHARDT
L. F.
(
1987
).
Neurite outgrowth onmuscle cell surfaces involves extracellular matrix receptors aswell asCa-dependent and independent cell adhesion molecules.
Proc. natn. Acad. Sci. U.S.A.
84
,
2555
2559
BOZYCZKO
D.
,
HORWITZ
A. F.
(
1986
).
The participation of a putative cell surface receptor forlaminin and fibronectin in peripheral neurite extension.
J. Neurosci.
6
,
1241
1251
BURKIDGE
K.
,
FERAMISCO
J. R.
(
1982
).
Alpha-actinin and vinculin from non-muscle cells: calcium-sensitive interactions with actin.
Cold SpringsHarbor Symp. quant. Biol.
46
,
587
597
BURRIDGE
K.
,
KELLY
T.
,
MANGEAT
P.
(
1982
).
Nonerythrocyte spectrins: Actin-membrane attachment proteins occurringin many cell types.
J. Cell Biol.
95
,
478
486
BURRIDGE
K.
,
MOLONY
L.
,
KELLY
T.
(
1987
).
Adhesion plaques; sites of transmembrane interaction between theextracellular matrix and theactin cytoskeleton.
J. Cell Sci.
(
Suppl. 8
) pp.
211
230
BUCK
C.A.
,
HORWITZ
A.F.
(
1987
).
Integrin, a transmembrane glycoprotein complex mediating cell-substratum adhesion.
J. Cell Sci.
(
Suppl. 8
) pp.
231
250
CHANG
S.
,
RATHJEN
F. G.
,
RAPER
J. A.
(
1987
).
Extension of neurites onaxons is impaired by antibodies against specific neural cell surface glycoproteins.
J. Cell Biol.
104
,
355
362
CHUONG
C.-M.
,
CROSSIN
K. L.
,
EDELMAN
G. M.
(
1987
).
Sequential expression and differential function of multiple adhesion molecules during formation of cerebellar cortical layers.
J. Cell Biol.
104
,
331
342
EDELMAN
G.M.
(
1988
).
Morphoregulatory molecules.
Biochem.
27
,
3533
3543
FUJIWARA
K.
,
POLLARD
T. D.
(
1976
).
Fluorescent antibody localization of myosin inthecytoplasm, cleavage furrow and mitotic spindle of human cells.
J. Cell Biol.
79
,
268
275
GEIGER
B.
,
VOLK
T.
,
VOLBERG
T.
,
BENDORI
R.
(
1987
).
Molecular interactions in adherens-type junctions.
J. Cell Sci.
(
Suppl. 8
) pp.
251
272
GRUMET
M.
,
HOFFMAN
S.
,
EDELMAN
G. M.
(
1984
).
Two antigenically related neuronal CAMs of different specificities mediate neuron-neuron andneuron-glia adhesion.
Proc. natn. Acad. Sci. U.S.A.
81
,
267
271
HALEGOUA
S.
(
1987
).
Changes inthephosphorylation and distribution of vinculin during nerve growth factor induced neurite outgrowth.
Devi Biol.
121
,
97
104
HAMMAMACK
J. A.
,
MCCARTHY
J. B.
,
PALM
S. L.
,
FURCHT
L. T.
,
LETOURNEAU
P.C.
(
1988
).
Growth cone guidanceby substrate-bound laminin pathways iscorrelated with neuron-to-pathway adhesivity.
Devi Biol
126
,
29
39
HATTA
K.
,
OKADA
T. S.
,
TAKEICHI
M.
(
1985
).
A monoclonal antibody disrupting calcium-dependent cell-cell adhesionof brain tissues. Possible role of its target antigen in animal pattern formation.
Proc. natn. Acad. Sci. U.S.A.
82
,
2789
2793
HATTA
K.
,
TAKAGI
S.
,
FUJISAWA
H.
,
TAKEICHI
M.
(
1987
).
Spatial and temporal expression pattern of N-cadherin adhesion molecules correlated with morphogenetic processes ofchicken embryos.
Devi Biol.
120
,
215
227
HIRANO
S.
,
NOSE
A.
,
HATTA
K.
,
KAWAKAMI
A.
,
TAKEICHI
M.
(
1987
).
Calcium-dependent cell-cell adhesion molecules (cadherins): subclass specificities and possible involvementof actin bundles.
J. Cell Biol.
105
,
2501
2510
HORWITZ
A.
,
DUGGAN
K.
,
BUCK
C.
,
BECKERLE
M.
,
BURJUDGE
K.
(
1986
).
Interaction of plasma membrane fibronectin receptor with talin-a transmembrane linkage.
Nature, Lond.
320
,
531
533
HYNES
R.O.
(
1987
).
Integrins: a family ofcell surface receptors.
Cell
48
,
549
554
JOCKUSCH
H.
,
JOCKUSCH
B. M.
(
1981
).
Structural proteins in the growth cone ofcultured spinal cord neurons.
Expl Cell Res.
131
,
345
352
KEILHAUSER
G.
,
FAISSNER
A.
,
SCHACHNER
M.
(
1985
).
Differential inhibition of neurone-neurone, neurone-astrocyte, and astrocyte-astrocyte adhesion byLI, L2,and N-CAM antibodies.
Nature, Lond.
316
,
728
730
KOENIG
E.
,
KINSMAN
S.
,
REPASKY
E.
,
SULTZ
L.
(
1985
).
Rapid mobility of motile varicosities and inclusions containinga-spectrin, actin and calmodulin in regenerating axons invitro.
J. Neurosci.
5
,
715
729
KOENIG
E.
,
REPASKY
E.
(
1985
).
A regional analysis of a-spectrin i n the isolated Mauthner neuron and inisolated axons of the goldfish and rabbit.
J. Neurosci.
5
,
705
714
KUCZMARSKI
E. R.
,
ROSENBAUM
J. L.
(
1979
).
Studies on the organization and localization of actin and myosin inneurons.
J. Cell Biol.
80
,
356
371
LAGENAUR
C.
,
LEMMON
V.
(
1987
).
An Ll-like molecule, the 8D9 antigen, isa potent substrate forneurite extension.
Proc. natn. Acad. Sci. U.S.A.
84
,
7753
7757
LEMMON
V.
,
MCLOON
S. C.
(
1986
).
The appearance of anLl-like molecule inthe chick primary visual pathway.
J. Neurosci.
6
,
2987
2994
LEMMON
V.
,
STAROS
E. B.
,
PERRY
H. E.
,
GOTTLIEB
D. I.
(
1982
).
Amonoclonal antibody which binds tothe surfaceof chick brain cells and myotubes: cell selectivity and the properties of the antigen.
Dev.Brain Res.
3
,
349
360
LETOURNEAU
P.C.
(
1975
).
Possible roles for cell-to-substratum adhesion in neuronal morphogenesis.
Devi Biol.
44
,
77
91
LETOURNEAU
P.C.
(
1979
).
Cell-substratum adhesion of neurite growth cones and its role inneurite elongation.
Expl Cell Res.
124
,
127
138
LETOURNEAU
P.C.
(
1981
).
Immunocytochemical evidencefor colocalization in neurite growth cones of actin and myosin and their relationship tocell-substratum adhesion.
Devi Biol.
85
,
113
122
LETOURNEAU
P.C.
(
1983
).
Differences in the organization ofactin i n the growth cones compared with the neurites of cultured neurons from chick embryos.
J. Cell Biol.
97
,
963
973
LETOURNEAU
P.C.
(
1985
).
Axonal growth and guidance.
In
MolecularBasesofNeural Development
(ed.
Edelman
G.M.
,
Gall
W. E.
,
Cowan
W. M.
), pp.
269
294
.
NewYork
:
John Wiley and Sons
.
LETOURNEAU
P. C.
,
PECH
I. V.
,
ROGERS
S. R.
,
PALM
S. L.
,
MCCARTHY
J. B.
,
FURCHT
L.T.
(
1988
).
Growth cone migration across extracellular components depends on integrin, but migration across glioma cells does not.
J.Neurosci. Res.
(in press).
LETOURNEAU
P. C.
,
SHATTUCK
T. A.
,
RESSLER
A. H.
(
1986
).
Branching ofsensory and sympathetic neurites is inhibitedby treatment with taxol.
J. Neurosci.
6
,
1912
1917
LEVINE
J.
,
WILLARD
M.
(
1981
).
Fodrin: axonally transported polypeptides associated with the internal periphery of many cells.
J. Cell Biol.
90
,
631
643
LUDUENA
M. A.
(
1973
).
Nerve cell differentiation invitro.
Devi Biol.
33
,
268
284
MARSH
L.
,
LETOURNEAU
P.C.
(
1984
).
Growth of neurites without filopodial or lamellipodial activity inthe presence of cytochalasin B.
J. Cell Biol.
99
,
2041
2047
MATSUMARA
F.
,
YAMASHIRO-MATSUMURA
S.
,
LIN
J.-C.
(
1983
).
Isolation and characterization of tropomyosin-containing micTofilaments from cultured cells.
J.biol. Chem.
258
,
6636
6644
PALM
S.L.
,
FURCHT
L.T.
(
1983
).
The production of laminin and fibronectin by Schwannoma cells: Cell-protein interactionsin vitro, and protein localization in peripheral nerve in vivo.
J. Cell Biol.
96
,
1218
1226
PENG
I.
,
BINDER
L. I.
,
BLACK
M. M.
(
1986
).
Biochemical and immunological analyses ofcytoskeletal domains of neurons.
J. Cell Biol.
102
,
252
262
PFENNINGER
K. H.
,
MAYUE-PFENNINGER
M. F.
(
1981
).
Lectin labeling ofsprouting neurons. II.Relative movement and appearance ofglyconjugates during plasmalemmal expansion.
J. Cell Biol.
89
,
547
559
PFENNINGER
K. H.
,
ELLIS
L.
,
JOHNSON
M. P.
,
FRIEDMAN
L. B.
,
SOMLO
S.
(
1983
).
Nerve growth cones isolated from fetalrat brain. I. Subcellular fractionation and characterization.
Cell
35
,
573
584
POLLARD
T.D.
,
COOPER
J. A.
(
1986
).
Actin and actin-binding proteins. Acritical evaluation ofmechanisms and functions.
Ann. Rev.Biochem.
55
,
987
1035
POLLERBERG
G. E.
,
BURRJDGE
K.
,
KREBS
K. E.
,
GOODMAN
S. R.
,
SCHACHNER
M.
(
1987
).
The 180-kD component ofthe neural cell adhesion molecule isinvolved incell contacts and cytoskeleton-membrane interactions.
Cell TissueRes.
250
,
227
236
POLLERBERG
G. E.
,
SADOUL
R.
,
GOURIDIS
C.
,
SCHACHNER
M.
(
1985
).
Selective expression ofthe 180-kD component of neural cell adhesion molecule N-CAM during development.
J. CellBiol.
101
,
1921
1929
POLLERBERG
G. E.
,
SCHACHNER
M.
,
DAVOUST
J.
(
1986
).
Differentiation state-dependent surface mobilities oftwo forms of the neural cell adhesion molecule.
Nature, Lond.
324
,
462
465
RAPER
J. A.
,
BASTIANI
M. J.
,
GOODMAN
C. S.
(
1983a
).
Pathfinding by neuronal growth cones in grasshopper embryos. I. Divergent choices made by thegrowth cones of sibling neurons.
J. Neurosci.
3
,
20
30
RAPER
J. A.
,
BASTIANI
M. J.
,
GOODMAN
C. S.
(
19836
).
Pathfinding by neuronal growth cones ingrasshopper embryos. II. Selective fasciculation onto specific axonal pathways.
J. Neurosci.
3
,
31
41
RAPER
J. A.
,
BASTIANI
M. J.
,
GOODMAN
C. S.
(
1984
).
Pathfinding by neuronal growth cones ingrasshopper embryos. IV. Theeffects ofablating the AandP axons upon behaviour of the Ggrowth cone.
J. Neurosci.
4
,
2329
2345
RATHJEN
F. J.
,
SCHACHNER
M.
(
1984
).
Immunocytochemicaland biochemical characterization ofa newneuronal cell surface component (LI antigen) which isinvolved incell adhesion.
EMBO J.
3
,
1
10
RATHJEN
F. G.
,
WOLFF
J. M.
,
FRANK
R.
,
BONHOEFFER
F.
,
RUTISHAUSER
U.
(
1987
).
Membrane glycoproteins involvedin neurite fasciculation.
J. Cell Biol.
104
,
343
353
ROGERS
S. L.
,
LETOURNEAU
P. C.
,
PALM
S. L.
,
MCCARTHY
J. B.
,
FURCHT
L.T.
(
1983
).
Neurite extension by peripheral and central nervous system neurons in response to substratum-bound fibronectin and laminin.
Devi Biol.
98
,
212
220
ROISEN
F.
,
INCZEDY-MARCSEK
M.
,
HSU
L.
,
YORKE
W.
(
1978
).
Myosin: Immunofiuorescent localization in neuronal and ghal cultures.
Science
199
,
1445
1448
SHAW
G.
,
OSBORN
M.
,
WEBER
K.
(
1981
).
Arrangement of neurofilaments, microtubules and microfilament-associated proteins incultured dorsal root ganglia cells.
Eur. J. CellBiol.
24
,
20
27
SHEIR-NEISS
G.
,
LAI
M. H.
,
MORRIS
N. R.
(
1978
).
Identification of a gene for β-tubulin in Aspergillus nidulans.
Cell
15
,
639
647
SMALL
R. P.
,
BLANK
M.
,
GHEZ
R.
,
PFENNINGER
K. H.
(
1984
).
Components of the plasma membrane ofgrowing axons.II. Diffusion of membrane protein complexes.
J. Cell Biol.
98
,
1434
1443
STALLCUP
W. B.
,
BEASLEY
L.
(
1985
).
Involvement ofthe nerve-growth factor-inducible large external glycoprotein (NILE)i n neurite fasciculation inprimary cultures ofratbrain.
Proc.natn. Acad. Sci. U.S.A.
82
,
1276
1280
STOSSEL
T.P.
(
1984
).
Contribution ofactin tothe structure of the cytoplasmic matrix.
J. CellBiol.
99
,
15s
22s
TAKEICHI
M.
(
1988
).
The cadherins: cell-cell adhesion molecules controlling animal morphogenesis.
Development
102
,
639
656
TOMASEK
J. J.
,
HAY
E. D.
,
FUJIWARA
K.
(
1982
).
Collagen modulates cell shape and cytoskeleton of embryo corneal and fibroma fibroblasts: Distribution of actin, a-actinin, and myosin.
Devi Biol.
92
,
107
122
TOMASELU
K.
,
NEUGEBAUER
K. M.
,
BIXBY
J. L.
,
LILIEN
J.
,
REICHARDT
L. F.
(
1988
).
N-cadherin and integrins: two receptor systems that mediate neuronal process outgrowth on astrocyte surfaces.
Neuron
1
,
33
43
TOMASELLI
K. J.
,
REICHARDT
L. F.
,
BIXBY
J. L.
(
1986
).
Distinct molecular interactions mediate neuronal process outgrowth on non-neuronal cell surfaces and extracellular matrices.
J. Cell Biol.
103
,
2659
2672
TOSNEY
K. W.
,
LANDMESSER
L. T.
(
1985
).
Development of the major pathways for neurite outgrowth in the chick limb.
Devi Biol.
109
,
193
214
TOWBIN
H.
,
STAEHELIN
T.
,
GORDON
J.
(
1979
).
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications.
Proc. natn. Acad. Sci. U.S.A.
76
,
4350
4354
VALLEE
R. B.
,
BLOOM
G. S.
,
THEURKAUF
W. E.
(
1984
).
Microtubule-associated proteins: subunits of the cytomatrix.
J. Cell Biol.
99
,
38s
46s
VAN DEN POL
A. N.
,
Di PORZIO
U.
,
RUTISHAUSER
U.
(
1986
).
Growth cone localization of neural cell adhesion molecule on central nervous system neurones in vitro.
J. Cell Biol.
102
,
2281
2294
This content is only available via PDF.