We have used a quantitative cell attachment assay to compare the interactions of cranial and trunk neural crest cells with the extracellular matrix (ECM) molecules fibronectin, laminin and collagen types I and IV. Antibodies to the beta 1 subunit of integrin inhibited attachment under all conditions tested, suggesting that integrins mediate neural crest cell interactions with these ECM molecules. The HNK-1 antibody against a surface carbohydrate epitope under certain conditions inhibited both cranial and trunk neural crest cell attachment to laminin, but not to fibronectin. An antiserum to alpha 1 intergrin inhibited attachment of trunk, but not cranial, neural crest cells to laminin and collagen type I, though interactions with fibronectin or collagen type IV were unaffected. The surface properties of trunk and cranial neural crest cells differed in several ways. First, trunk neural crest cells attached to collagen types I and IV, but cranial neural crest cells did not. Second, their divalent cation requirements for attachment to ECM molecules differed. For fibronectin substrata, trunk neural crest cells required divalent cations for attachment, whereas cranial neural crest cells bound in the absence of divalent cations. However, cranial neural crest cells lost this cation-independent attachment after a few days of culture. For laminin substrata, trunk cells used two integrins, one divalent cation-dependent and the other divalent cation-independent (Lallier, T. E. and Bronner-Fraser, M. (1991) Development 113, 1069–1081). In contrast, cranial neural crest cells attached to laminin using a single, divalent cation-dependent receptor system. Immunoprecipitations and immunoblots of surface labelled neural crest cells with HNK-1, alpha 1 integrin and beta 1 integrin antibodies suggest that cranial and trunk neural crest cells possess biochemically distinct integrins. Our results demonstrate that cranial and trunk cells differ in their mechanisms of adhesion to selected ECM components, suggesting that they are non-overlapping populations of cells with regard to their adhesive properties.
REFERENCES
Belkin
V. M.
, Belkin
A. M.
, Koteliansky
V. E.
(1990
) Human smooth muscle VLA-1 integrin: Purification, substrate specificity, localization in aorta, and expression during development.
J. Cell Biol
111
, 2159
–2170
Bronner-Fraser
M.
(1985
) Alterations in neural crest migration by a monoclonal antibody that affects cell adhesion.
J. Cell Biol
101
, 610
–617
Bronner-Fraser
M.
(1986
) An antibody to a receptor for fibronectin and laminin perturbs cranial neural crest development in vivo.
Dev. Biol
117
, 528
–536
Bronner-Fraser
M.
(1987
) Perturbation of cranial neural crest migration by the HNK-1 antibody.
Dev. Biol
123
, 321
–331
Bronner-Fraser
M.
(1988
) Distribution and function of tenascin during cranial neural crest development in the chick.
J. Neurosci. Res
21
, 135
–147
Bronner-Fraser
M.
, Lallier
T.
(1988
) A monoclonal antibody against a laminin-heparin sulfate proteoglycan complex perturbs cranial neural crest migration in vivo.
J. Cell Biol
106
, 1321
–1329
Buck
C. A.
, Shea
E.
, Duggan
K.
, Horwitz
A. F.
(1986
) Integrin (the JG22 antigen): Functionality requires oligomeric integrity.
J. Cell Biol
103
, 2421
–2428
Burnette
W. M.
(1981
) Western Blotting: Electrophoretic transfer of protein from sodium dodecyl sulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
Anal. Biochem
112
, 195
–302
Carter
W. G.
, Wayner
E. A.
, Bouchard
T. S.
, Kaur
P.
(1990
) The role ofand in cell-cell and cell-substratum adhesion of human epidermal cells.
J. Cell Biol
110
, 1387
–1404
Charo
I. F.
, Nannizzi
L.
, Smith
J. W.
, Cheresh
D. A.
(1990
) The vitronectin receptorv binds fibronectin and acts in concert within promoting cellular attachment and spreading on fibronectin.
J. Cell Biol
111
, 2795
–2800
Cheresh
D. A.
, Pytela
R.
, Pierschbacher
M. D.
, Klier
F. G.
, Ruoslahti
E.
, Reisfeld
R. A.
(1987
) An arg-gly-asp-directed receptor on the surface of human melanoma cells exists in a divalent cation-dependent functional complex with the disialoganglioside GD2.
J. Cell Biol
105
, 1163
–1173
Dedhar
S.
, Saulnier
R.
(1990
) Alterations in integrin receptor expression on chemically transformed human cells: Specific enhancement of laminin and collagen receptor complexes.
J. Cell Biol
110
, 481
–491
Dedhar
S.
, Gray
V.
(1990
) Isolation of a novel integrin receptor mediating arg-gly-asp-directed cell adhesion to fibronectin and type I collagen from human neuroblastoma cells. Association of a novel-related subunit with v.
J. Cell Biol
110
, 2185
–2193
Duband
J. L.
, Thiery
J. P.
(1987
) Distribution of laminin and collagens during avian neural crest development.
Development
101
, 461
–478
Edwards
J. G.
, Robson
R. T.
, Campbell
G.
(1987
) A major difference between serum and fibronectin in the divalent cation requirement for adhesion and spreading of BHK21 cells.
J. Cell Sci
87
, 657
–665
Edwards
J. G.
, Hameed
H.
, Campbell
G.
(1988
) Induction of fibroblast spreading by Mn2+: a possible role for unusual binding sites for540divalent cations in receptors for proteins containing Arg-Gly-Asp.
J. Cell Sci
89
, 507
–513
Elices
M. J.
, Hemler
M. E.
(1989
) The human integrin VLA-2 is a collagen receptor on some cells and a collagen/laminin receptor on others.
Proc. Natl. Acad. Sci. USA
86
, 9906
–9910
Elices
M. J.
, Urry
L. A.
, Hemler
M. E.
(1991
) Receptor functions for the integrin VLA-3: fibronectin, collagen, and laminin binding are differentially influenced by ARG-LY-ASP peptide and by divalent cations.
J. Cell Biol
112
, 169
–181
Erickson
C. A.
(1988
) Control of pathfinding by the avian trunk neural crest.
Development
103
, 63
–80
Fauquet
M.
, Smith
J.
, Ziller
C.
, LeDouarin
N. M.
(1981
) Acetylcholine and catecholamine synthesis in neural crest cells cultured in vitro.
J. Neurosci
1
, 478
–492
Gailit
J.
, Ruoslahti
E.
(1988
) Regulation of the fibronectin receptor affinity by divalent cations.
J. Biol. Chem
263
, 12927
–12932
Hall
D. E.
, Reichardt
L. F.
, Crowley
E.
, Holley
B.
, Moezzi
H.
, Sonnenberg
A.
, Damsky
C. H.
(1990
) Theand integrin heterodimers mediate cell attachment to distinct sites on laminin.
J. Cell Biol
110
, 2175
–2184
Hamburger
V.
, Hamilton
H. L.
(1951
) A series of normal stages in the development of the chick embryo.
J. Morph
88
, 49
–92
Horwitz
A. F.
, Duggan
K.
, Greggs
R.
, Decker
C.
, Buck
C.
(1985
) The cell substratum attachment (JG22) antigen has properties of a receptor for laminin and fibronectin.
J. Cell Biol
101
, 2134
–2144
Howard
M.
, Bronner-Fraser
M.
(1985
) The influence of neural tube derived factors on differentiation of neural crest cells in vitro. Histochemical study on the appearance of adrenergic cells.
J. Neurosci
5
, 3302
–3309
Hynes
R. O.
(1987
) Integrins: a family of cell surface receptors.
Cell
48
, 549
–554
Ignatius
M. J.
, Reichardt
L. F.
(1988
) Identification of a neuronal laminin receptor: An Mr 200K/120K integrin heterodimer that binds laminin in a divalent cation-dependent manner.
Neuron
1
, 713
–725
Kahn
C. R.
, Coyle
J. T.
, Cohen
A. M.
(1980
) Head and trunk neural crest in vitro: Autonomic neuron differentiation.
Dev. Biol
77
, 340
–348
Kirchhofer
D.
, Languino
L. R.
, Ruoslahti
E.
, Pierschbacher
M.
(1990
) integrins from different cell types show different binding specificities.
J. Biol. Chem
265
, 615
–618
Kirchhofer
D.
, Grzesiak
J.
, Pierschbacher
M.
(1991
) Calcium as a potential physiological regulator of integrin-mediated cell adhesion.
J. Biol. Chem
266
, 4471
–4477
Kramer
R. H.
, Cheng
Y.-F.
, Clyman
R.
(1990
) Human microvascular endothelial cells useand integrin receptor complexes to attach to laminin.
J. Cell Biol
111
, 1233
–1243
Krotoski
D.
, Domingo
C.
, Bronner-Fraser
M.
(1986
) Distribution of a putative cell surface receptor for fibronectin and laminin in avian embryo.
J. Cell Biol
103
, 1061
–1071
Laemmli
U. K.
(1970
) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227
, 680
–685
Lallier
T. E.
, Bronner-Fraser
M.
(1991
) Avian neural crest cell adhesion to laminin: Involvement of divalent cation dependent and independent integrins.
Development
113
, 1069
–1081
Leblanc
G. G.
, Epstein
M. L.
, Bronner-Fraser
M.
(1990
) Differential development of cholinergic neurons from cranial and trunk neural crest cells in vitro.
Dev. Biol
137
, 318
–330
LeDouarin
N. M.
, Teillet
M. A.
(1974
) Experimental analysis of the migration and differentiation of neuroblasts of the autonomic nervous system and of neuroectodermal mesenchymal derivatives, using a biological cell marking technique.
Dev. Biol
41
, 162
–184
LeDouarin
N. M.
, Renaud
D.
, Teillet
M. A.
, Le Douarin
G. H.
(1975
) Cholinergic differentiation of presumptive adrenergic neuroblasts in interspecific chimeras after heterotopic transplantation.
Proc. Natl. Acad. Sci. USA
72
, 728
–732
Lee
E. D.
, Lotz
M. M.
, Steele
G. D.
, Mercurio
A. M.
(1992
) The integrinis a laminin receptor.
J. Cell Biol
117
, 671
–678
LeLievre
C. S.
, Schweizer
G. G.
, Ziller
C. M.
, LeDouarin
N. M.
(1980
) Restrictions of developmental capabilities in neural crest cellderivatives as tested by in vivo transplantation experiments.
Dev. Biol
77
, 362
–378
Lotz
M. M.
, Korzelius
C. A.
, Mercurio
A. M.
(1990
) Human colon carcinoma cells use multiple receptors to adhere to laminin: involvement ofand integrins.
Cell Regul
1
, 249
–257
McClay
D. R.
, Wessel
G. M.
, Marchase
R. B.
(1981
) Intercellular recognition: Quantitation of initial binding events.
Proc. Natl. Acad. Sci. USA
78
, 4975
–4979
Nakamura
H.
, Ayer-LeLievre
C. S.
(1982
) Mesoectodermal capabilities of the trunk neural crest of birds.
J. Embryol. Exp. Morph
70
, 1
–18
Newgreen
D. F.
, Thiery
J. P.
(1980
) Fibronectin in early avian embryos: Synthesis and distribution along the migration pathways of neural crest cells.
Cell Tissue Res
211
, 269
–291
Niu
M. C.
(1947
) The axial organization of the neural crest, studied with particular reference to its pigmentary component.
J. Exp. Zool
105
, 79
–114
Perris
R.
, Krotoski
D.
, Lallier
T.
, Domingo
C.
, Sorrell
J. M.
, Bronner-Fraser
M.
(1991
) Spatial and temporal changes in the distribution of proteoglycans during avian neural crest development.
Development
111
, 583
–599
Perris
R.
, Krotoski
D.
, Bronner-Fraser
M.
(1991
) Collagens in avian neural crest development: Distribution in vivo and migration-promoting ability in vitro.
Development
113
, 969
–984
Pomeranz
H. D.
, Gershon
M. D.
(1990
) Colonization of the avian hindgut by cells derived from the sacral neural crest.
Dev. Biol
137
, 378
–394
Rossino
P.
, Gavazzi
I.
, Timpl
R.
, Aumailley
M.
, Abbadini
M.
, Giancotti
F.
, Silengo
L.
, Marchisio
P. C.
, Tarone
G.
(1990
) Nerve growth factor induces ncreased expression of a laminin-binding integrin in rat phenochromocytoma PC12 cells.
Exp. Cell Res
189
, 100
–108
Serbedzija
G.
, Fraser
S. E.
, Bronner-Fraser
M.
(1991
) Vital dye labelling demonstrates a sacral neural crest contribution to the enteric nervous system of chick and mouse embryos.
Development
111
, 859
–866
Shaw
L. M.
, Messier
J. M.
, Mercurio
A. M.
(1990
) The activation dependent adhesion of macrophages to laminin involves cytoskeletal anchoring and phosphorylation of theintegrin.
J. Cell Biol
110
, 2167
–2174
Shimizu
Y.
, Van Seventer
G. A.
, Horgan
K. J.
, Shaw
S.
(1990
) Costimulation of proliferative responses of resting CD4+ T cells by the interaction of LA-4 and VLA-5 with fibronectin of VLA-6 with laminin.
J. Immunol
145
, 59
–67
Smith
J. W.
, Cheresh
D. A.
(1988
) The arg-gly-asp binding domain of the vitronectin receptor. Photoaffinity cross-linking implicates amino acid residues 61–203 of thesubunit.
J. Biol. Chem
263
, 18726
–18731
Sonnenberg
A.
, Modderman
P. W.
, Hogervorst
F.
(1988
) Laminin receptor on platelets is the integrin VLA-6.
Nature
336
, 487
–489
Sonnenberg
A.
, Linders
C. J. T.
, Modderman
P. W.
, Damsky
C. H.
, Aumailley
M.
, Timpl
R.
(1990
) Integrin recognition of different cell-binding fragments of laminin (P1, E3, E8) and evidence thatbut not functions as a major receptor for fragment E8.
J. Cell Biol
110
, 2145
–2155
Staatz
W. D.
, Rajpara
S. M.
, Wayner
E. A.
, Carter
W. G.
, Santoro
S. A.
(1989
) The membrane glycoprotein Ia-IIa (VLA-2) complex mediates the Mg++ -dependent adhesion of platelets to collagen.
J. Cell Biol
108
, 1917
–1924
Staatz
W. D.
, Walsh
J. J.
, Pexton
T.
, Santoro
S. A.
(1990
) The2 1integrin cell surface collagen receptor binds to the a1(I)-CB3 peptide of collagen.
J. Biol. Chem
265
, 4778
–4781
Tan
S. S.
, Crossin
K. L.
, Hoffman
S.
, Edelman
G. M.
(1987
) Asymmetric expression in somites of cytotactin and its proteoglycan ligand is correlated with neural crest cell distribution.
Proc. Natl. Acad. Sci. USA
84
, 7977
–7981
Teillet
M. A.
, Kalcheim
K.
, LeDouarin
N. M.
(1987
) Formation of the dorsal root ganglia in the avian embryo: segmental origin and migratory behavior of neural crest progenitor cells.
Dev. Biol
120
, 329
–347
Tomaselli
K. J.
, Damsky
C. H.
, Reichardt
L. F.
(1988
) Purification and characterization of mammalian integrins expressed by a rat neural cell line (PC12): Evidence that they function asheterodimeric receptors for laminin and type IV collagen.
J. Cell Biol
107
, 1241
–1252
Turner
D. C.
, Flier
L. A.
, Carbonetto
S.
(1987
) Magnesium-dependent attachment and neurite outgrowth by PC12 cells on collagen and laminin substrata.
Dev. Biol
121
, 510
–525
Vogel
B. E.
, Tarone
G.
, Giancotti
F. G.
, Gailit
J.
, Ruoslahti
E.
(1990
) A novel fibronectin receptor with an unexpected subunit composition (v).
J. Biol. Chem
265
, 5934
–5937
von Boxberg
Y.
, Wutz
R.
, Schwarz
U.
(1990
) Use of the biotin-avidin system for the labelling, isolation and characterization of neural cell surface proteins.
Eur. J. Biochem
190
, 249
–256
Wayner
E. A.
, Carter
W. G.
(1987
) Identification of multiple cell adhesion receptors for type IV collagen and fibronectin in human fibrosarcoma cells possessing uniqueand subunits.
J. Cell Biol
105
, 1873
–1884
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