Cultured rat Schwann cells were stimulated to deposit fibrillar extracellular matrix by treatment with ascorbic acid in the absence of nerve cells. Immunofluoresence staining of the matrix showed that it contains collagens types I and IV, fibronectin and perlecan but not laminin. Collagen type IV, fibronectin and perlecan co-distributed completely in the matrix fibrils, whereas collagen type I was present in only a subset of these fibrils. Time course studies indicated that collagen type I fibrils appear at late stages of matrix formation. Digestion of Schwann cell extracellular matrix with collagenase effectively disrupted most of the matrix including fibronectin fibrils. This was in contrast with fibroblasts, where collagenase treatment removed collagen with no visible effect on fibronectin fibrils. alpha5 integrin was expressed on the cell surface of Schwann cells and partially codistributed with fibronectin-containing fibrils. This suggests that the inability of Schwann cells to deposit fibronectin-containing matrix through a conventional, collagen-independent mechanism was not due to the lack of fibronectin-binding integrins on their cell surface. Polyclonal anti-fibronectin antibodies inhibited the deposition of fibronectin into the matrix fibrils, whereas collagen type IV fibrils were generally unaffected. Growth of Schwann cells on collagen type IV-coated substrate in the absence of ascorbate induced deposition of fine fibronectin fibrils. These results suggest that Schwann cells use an apparently novel, collagen type IV-dependent mechanism for the deposition of fibronectin into their extracellular matrix.

REFERENCES

Akiyama
S. K.
,
Yamada
S. S.
,
Chen
W.-T.
,
Yamada
K. M.
(
1989
).
Analysis of fibronectin receptor function with monoclonal antibodies: roles in cell adhesion, migration, matrix assembly, and cytoskeletal organization.
J. Cell Biol
109
,
863
875
Baron-VanEvercooren
A.
,
Kleinman
H. K.
,
Seppa
H. E. J.
,
Rentier
B.
,
Dubois-Dalcq
M.
(
1982
).
Fibronectin promotes rat Schwann cell growth and motility.
J. Cell Biol
93
,
211
216
Baron-VanEvercooren
A.
,
Gansmuller
A.
,
Gumpel
M.
,
Baumann
N.
,
Kleinman
H. K.
(
1986
).
Schwann cell differentiation in vitro: extracellular matrix deposition and interaction.
Dev. Neurosci
8
,
182
196
Bunge
M. B.
,
Williams
A. K.
,
Wood
P. M.
,
Uitto
U.
,
Jeffrey
J. J.
(
1980
).
Comparison of nerve cell and nerve cell plus Schwann cell cultures, with particular emphasis on basal lamina and collagen formation.
J. Cell Biol
84
,
184
202
Bunge
M. B.
,
Williams
A. K.
,
Wood
P. M.
(
1982
).
Neuron-Schwann cell interaction in basal lamina formation.
Dev. Biol
92
,
449
460
Carey
D. J.
,
Todd
M. S.
,
Rafferty
M. C.
(
1986
).
Schwann cell myelination: induction by exogenous basement membrane-like extracellular matrix.
J. Cell Biol
102
,
2224
2263
Chen
H.
,
Mosher
D. F.
(
1996
).
Formation of sodium dodecyl sulfate-stable fibronectin multimers: Failure to detect products of thiol-disulfide exchange in cyanogen bromide or limited acid digests of stabilized matrix fibronectin.
J. Biol Chem
271
,
9084
9089
Chen
L. B.
,
Murray
A.
,
Segal
R. A.
,
Bushnell
A.
,
Walsh
M. L.
(
1978
).
Studies on intercellular LETS glycoprotein matrices.
Cell
14
,
377
391
Chernousov
M. A.
,
Faerman
A. I.
,
Frid
M. G.
,
Printseva
O. Yu.
,
Koteliansky
V. E.
(
1987
).
Monoclonal antibody to fibronectin which inhibits extracellular matrix assembly.
FEBS Lett
217
,
124
128
Chernousov
M. A.
,
Fogerty
F. J.
,
Koteliansky
V. E.
,
Mosher
D. F.
(
1991
).
Role of the I-9 and III-1 modules of fibronectin in formation of an extracellular fibronectin matrix.
J. Biol. Chem
266
,
10851
10858
Chernousov
M. A.
,
Stahl
R. C.
,
Carey
D. J.
(
1996
).
Schwann cells secrete a novel, collagen-like adhesive protein that binds N-syndecan.
J. Biol. Chem
271
,
13844
13853
Choi
M. G.
,
Hynes
R. O.
(
1979
).
Biosynthesis and processing of fibronectin in NIL. 8 hamster cells.
J. Biol. Chem
254
,
12050
12055
Cornbrooks
C. J.
,
Carey
D. J.
,
McDonald
J. A.
,
Timpl
R.
,
Bunge
R. P.
(
1983
).
In vivo and in vitro observations on laminin production by Schwann cells.
Proc. Nat. Acad. Sci. USA
80
,
3850
3854
Darribere
T.
,
Koteliansky
V. E.
,
Chernousov
M. A.
,
Akiyama
S. K.
,
Yamada
K. M.
,
Thiery
J. P.
,
Boucaut
J.-C.
(
1992
).
Distinct regions of human fibronectin are essential for fibril assembly in an in vivo developing system.
Dev. Dynam
194
,
63
70
DeClerck
Y. A.
,
Jones
P. A.
(
1980
).
The effect of ascorbic acid on the nature and production of collagen and elastin by rat smooth-muscle cells.
Biochem. J
186
,
217
225
Dzamba
B. J.
,
Peters
D. M.
(
1991
).
Arrangement of cellular fibronectin in noncollagenous fibrils in human fibroblast cultures.
J. Cell Sci
100
,
605
612
Dzamba
B. J.
,
Wu
H.
,
Jaenisch
R.
,
Peters
D. M.
(
1993
).
Fibronectin binding site in type I collagen regulates fibronectin fibril formation.
J. Cell Biol
121
,
1165
1172
Fogerty
F. J.
,
St. Akiyama
K.
,
Yamada
K. M.
,
Mosher
D. F.
(
1990
).
Inhibition of binding of fibronectin to matrix assembly sites by anti-integrin (5 1) antibodies.
J. Cell Biol
111
,
699
708
Furcht
L. T.
,
Smith
D.
,
Wendelschafer-Crabb
G.
,
Mosher
D. F.
,
Foidart
J. M.
(
1980
).
Fibronectin presence in native collagen fibrils of human fibroblasts: immunoperoxidase and immunoferritin localization.
J. Histochem. Cytochem
28
,
1319
1333
Ide
C.
(
1996
).
Peripheral nerve regeneration.
Neurosci. Res
25
,
101
121
Koteliansky
V. E.
,
Arsenyeva
E. L.
,
Bogacheva
G. T.
,
Chernousov
M. A.
,
Glukhova
M. A.
,
Ibraghimov
A. R.
,
Metsis
M. L.
,
Petrosyan
M. N.
,
Rokhlin
O. V.
(
1982
).
Identification of the species-specific antigenic determinant(s) of human plasma fibronectin by monoclonal antibodies.
FEBS Lett
142
,
199
202
Lefcort
F.
,
Venstrom
K.
,
McDonald
J. A.
,
Reichardt
L. F.
(
1992
).
Regulation of expression of fibronectin and its receptor,5 1 during development and regeneration of peripheral nerve.
Development
116
,
767
782
McDonald
J. A.
,
Kelley
D. G.
,
Broekelmann
T. J.
(
1982
).
Role of fibronectin in collagen deposition: Fab' to the gelatin-binding domain of fibronectin inhibits both fibronectin and collagen organization in fibroblast extracellular matrix.
J. Cell Biol
92
,
485
492
McDonald
J. A.
(
1994
).
Extracellular matrix assembly.
Meth. Enzymol
245
,
518
531
McGarvey
M. L.
,
Baron-VanEvercooren
A.
,
Kleinman
H. K.
,
Dubois-Dalcq
M.
(
1984
).
Synthesis and effects of basement membrane components in cultured rat Schwann cells.
Dev. Biol
105
,
18
28
McKeown-Longo
P. J.
,
Mosher
D. F.
(
1983
).
Binding of plasma fibronectin to cell layers of human skin fibroblasts.
J. Cell Biol
97
,
466
472
McKeown-Longo
P. J.
,
Mosher
D. F.
(
1984
).
Mechanism of formation of disulfide-bonded multimer of plasma fibronectin in cell layers of cultured human fibroblasts.
J. Biol. Chem
259
,
12210
12215
Mosher
D. F.
,
Sottile
J.
,
Wu
C.
,
McDonald
J. A.
(
1992
).
Assembly of extracellular matrix.
Curr. Opin. Cell Biol
4
,
810
818
Murad
S.
,
Tajima
S.
,
Johnson
G. R.
,
Sivarajah
A.
,
Pinnell
S. R.
(
1983
).
Collagen synthesis in cultured human skin fibroblasts: effect of ascorbic acid and its analogs.
J. Invest. Dermatol
81
,
158
162
Peltonen
J.
,
Jaakkola
S.
,
Chu
M.-L.
,
Uitto
J.
(
1990
).
Selective expression of extracellular matrix genes encoding type VI collagen and laminin by Schwann cells, perineurial cells, and fibroblasts from normal nerve and neurofibromas.
Ann. NY Acad. Sci
580
,
501
504
Vaheri
A.
,
Kurkinen
M.
,
Lehto
V.-P.
,
Linder
E.
,
Timpl
R.
(
1978
).
Codistribution of pericellular matrix proteins in cultured fibroblasts and loss in transformation: fibronectin and procollagen.
Proc. Nat. Acad. Sci. USA
75
,
4944
4948
Wu
C.
,
Bauer
J. S.
,
Juliano
R. L.
,
McDonald
J. A.
(
1993
).
The5 1integrin fibronectin receptor, but not the5cytoplasmic domain, functions in an early and essential step in fibronectin matrix assembly.
J. Biol. Chem
268
,
21883
21888
This content is only available via PDF.