The outgrowth of parietal endoderm (PE) cells from precursor endodermal cells is one of the first differentiation events that occur in mouse embryos. We have analyzed the molecular determinants of this process by placing isolated inner cell masses (ICMs) on defined extracellular matrix substrata in microdrop cultures. Differentiation and outgrowth of PE required a fibronectin substratum. Laminin supported the adhesion and outgrowth of visceral endoderm (VE) and actively suppressed the differentiation of PE in mixtures of fibronectin and laminin. Collagen type IV, gelatin, vitronectin or entactin supported little or no endodermal outgrowth. Trophectoderm (TE) cells have been implied to be important in PE induction in vivo. We found that recombination of ICMs in culture with TE cells, or with medium conditioned by TE cells, greatly increased the differentiation of PE. TE cells stimulated PE outgrowth on substrata other than fibronectin. One cytokine secreted by trophoblast and endodermal cells, parathyroid hormone-related peptide (PTHrP), was critical for outgrowth on any substratum. A function-perturbing antibody to PTHrP reduced the number of PE cells, whereas the addition of PTHrP increased that number. Furthermore, addition of PTHrP changed the substratum requirements for outgrowth, making laminin, vitronectin and low concentrations of fibronectin permissive for PE outgrowth. Immunostaining with anti-integrin antibodies showed that fully differentiated PE cells outgrowing on fibronectin expressed alpha 5, alpha 6 and alpha v beta 3 integrins. However, analysis of outgrowths in the presence of function-perturbing antibodies to alpha 5, alpha 6 and alpha v beta 3 integrins showed that these integrins directed PE outgrowth only on fibronectin, laminin and vitronectin substrata, respectively. We have shown that there is a cooperative interplay of extracellular matrix, integrins and PTHrP that modulates PE outgrowth.

Reference

Adams
J. C.
,
Watt
F. M.
(
1993
)
Regulation of development and differentiation by the extracellular matrix.
Development
117
,
1183
1198
Albano
R. M.
,
Groome
N.
,
Smith
J. C.
(
1993
)
Activins are expressed in preimplantation mouse embryos and in ES and EC cells and are regulated on their differentiation.
Development
117
,
711
723
Beauvais
A.
,
Erickson
C. A.
,
Goins
T.
,
Craig
S. E.
,
Humphries
M. J.
,
Thiery
J. P.
,
Dufour
S.
(
1995
)
Changes in the fibronectin-specific integrin expression pattern modify the migratory behavior of sarcoma S180 cells in vitro and in the embryonic environment.
J. Cell Biol
128
,
699
713
Behrendtsen
O.
,
Alexander
C. M.
,
Werb
Z.
(
1992
)
Metalloproteinases mediate extracellular matrix degradation by cells from mouse blastocyst outgrowths.
Development
114
,
447
456
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
Calof
A. L.
,
Lander
A. D.
(
1991
)
Relationship between neuronal migration and cell-substratum adhesion: laminin and merosin promote olfactory neuronal migration but are anti-adhesive.
J. Cell Biol
115
,
779
794
Carnegie
J. A.
(
1991
)
Immunolocalization of fibronectin and laminin within rat blastocysts cultured under serum-free conditions.
J. Reprod. Fert
91
,
423
434
Casanova
J. E.
,
Grabel
L. B.
(
1988
)
The role of cell interactions in the differentiation of teratocarcinoma-derived parietal and visceral endoderm.
Dev. Biol
129
,
124
139
Chan
S. D. H.
,
Strewler
G. J.
,
King
K. L.
,
Nissenson
R. A.
(
1990
)
Expression of a parathyroid hormone-like protein and its receptor during differentiation of embryonal carcinoma cells.
Mol. Endocrinol
4
,
638
646
Colosi
P.
,
Talamantes
F.
,
Linzer
D. I.
(
1987
)
Molecular cloning and expression of mouse placental lactogen I complementary deoxyribonucleic acid.
Mol. Endocrinol
1
,
767
776
Coopman
P. J.
,
Bracke
M. E.
,
Lissitzky
J. C.
,
De Bruyne
G. K.
,
Van Roy
F. M.
,
Foidart
J. M.
,
Mareel
M. M.
(
1991
)
Influence of basement membrane molecules on directional migration of human breast cell lines in vitro.
J. Cell Sci
98
,
395
401
Cross
J. C.
,
Flannery
M. L.
,
Blanar
M. A.
,
Steingrimsson
E.
,
Jenkins
N. A.
,
Copeland
N. G.
,
Rutter
W. J.
,
Werb
Z.
(
1995
)
Hxt encodes a basic helix-loop-helix transcription factor that regulates trophoblast cell development.
Development
121
,
2513
2523
Damsky
C. H.
,
Richa
J.
,
Solter
D.
,
Knudsen
K.
,
Buck
C.
(
1983
)
Identification and purification of a cell surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue.
Cell
34
,
455
466
Damsky
C. H.
,
Werb
Z.
(
1992
)
Signal transduction by integrin receptors for extracellular matrix: cooperative processing of extracellular information.
Curr. Opin. Cell Biol
4
,
772
781
Dziadek
M.
,
Adamson
E.
(
1978
)
Localization and synthesis of alphafoetoprotein in post-implantation mouse embryos.
J. Embryol. Exp. Morph
43
,
289
313
Enders
A. C.
,
Given
R. L.
,
Schlafke
S.
(
1978
)
Differentiation and migration of endoderm in the rat and mouse at implantation.
Anat. Rec
190
,
65
77
Enders
A. C.
,
Lantz
K. C.
,
Schlafke
S.
(
1990
)
Differentiation of the inner cell mass of the baboon blastocyst.
Anat. Rec
226
,
237
248
ffrench-Constant
C.
,
Hollingsworth
A.
,
Heasman
J.
,
Wylie
C. C.
(
1991
)
Response to fibronectin of mouse primordial germ cells before, during and after migration.
Development
113
,
1365
1373
Fujii
K.
,
Dousaka-Nakajima
N.
,
Imamura
S.
(
1995
)
Epidermal growth factor enhancement of HSC-1 human cutaneous squamous carcinoma cell adhesion and migration on type I collagen involves selective up-regulation of alpha 2 beta 1 integrin expression.
Exp. Cell Res
216
,
261
272
Gardner
R. L.
(
1982
)
Investigation of cell lineage and differentiation in the extraembryonic endoderm of the mouse embryo.
J. Embryol. Exp. Morph
68
,
175
198
Gardner
R. L.
,
Davies
T. J.
,
Carey
M. S.
(
1988
)
Effect of delayed implantation on differentiation of the extra-embryonic endoderm in the mouse blastocyst.
Placenta
9
,
343
359
George
E. L.
,
Georges-Labouesse
E. N.
,
Patel-King
R. S.
,
Rayburn
H.
,
Hynes
R. O.
(
1993
)
Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin.
Development
119
,
1079
1091
Glass
R. H.
,
Aggeler
J.
,
Spindle
A.
,
Pedersen
R. A.
,
Werb
Z.
(
1983
)
Degradation of extracellular matrix by mouse trophoblast outgrowths: A model for implantation.
J. Cell Biol
96
,
1108
1116
Grabel
L. B.
,
Watts
T. D.
(
1987
)
The role of extracellular matrix in the migration and differentiation of parietal endoderm from teratocarcinoma embryoid bodies.
J. Cell Biol
105
,
441
448
Grover
A.
,
Edwards
S. A.
,
Bourdon
M.
,
Adamson
E. D.
(
1987
)
Proteoglycan 19, laminin and collagen type IV production is correlated with the levels of mRNA in F9 cell aggregates differentiating in the presence or absence of cyclic AMP.
Differentiation
36
,
138
144
Hierck
B. P.
,
Thorsteinsdottir
S.
,
Niessen
C. M.
,
Freund
E.
,
Iperen
L. V.
,
Feyen
A.
,
Hogervorst
F.
,
Poelmann
R. E.
,
Mummery
C. L.
,
Sonnenberg
A.
(
1993
)
Variants of the6 1laminin receptor in early murine development: distribution, molecular cloning and chromosomal localization of the mouse integrin6 subunit.
CellAdhes. Commun
1
,
33
53
Hogan
B. L. M.
,
Newman
R.
(
1984
)
A scanning electron microscope study of the extraembryonic endoderm of the 8th-day mouse embryo.
Differentiation
26
,
138
143
Hogan
B.
,
Tilly
R.
(
1978
)
In vitro development of inner cell masses isolated immunosurgically from mouse blastocysts. II. Inner cell masses from 3. 5-to 4. 0-day p. c. blastocysts.
J. Embryol. Exp. Morph
45
,
107
121
Hogan
B. L. M.
,
Tilly
R.
(
1981
)
Cell interactions and endoderm differentiation in cultured mouse embryos.
J. Embryol. Exp. Morph
62
,
379
394
Hogan
B. L. M.
,
Cooper
A. R.
,
Kurkinen
M.
(
1980
)
Incorporation into Reichert's membrane of laminin-like extracellular proteins synthesized by parietal endoderm cells of the mouse embryo.
Dev. Biol
80
,
289
300
Hogan
B. L. M.
,
Taylor
A.
,
Cooper
A. R.
(
1982
)
Murine parietal endoderm cells synthesise heparan sulphate and 170K and 145K sulphated glycoproteins as components of Reichert's membrane.
Dev. Biol
90
,
210
214
Imada
M.
,
Imada
S.
,
Iwasaki
H.
,
Kume
A.
,
Yamaguchi
H.
,
Moore
E. E.
(
1987
)
Fetomodulin: marker surface protein of fetal development which is modulatable by cyclic AMP.
Dev. Biol
122
,
483
491
Jackson
B. W.
,
Grund
C.
,
Winter
S.
,
Franke
W. W.
,
Illmensee
K.
(
1981
)
Formation of cytoskeletal elements during mouse embryogenesis. II. Epithelial differentiation and intermediate-sized filaments in early postimplantation embryos.
Differentiation
20
,
203
216
Jiang
R.
,
Grabel
L. B.
(
1995
)
Function and differential regulation of the6 integrin isoforms during parietal endoderm differentiation.
Exp. Cell Res
217
,
195
204
Karaplis
A. C.
,
Luz
A.
,
Glowacki
J.
,
Bronson
R. T.
,
Tybulewicz
V. L.
,
Kronenberg
H. M.
,
Mulligan
R. C.
(
1994
)
Lethal skeletal dysplasia from targeted disruption of the parathyroid hormone-related peptide gene.
Genes Dev
8
,
277
289
Klemke
R. L.
,
Yebra
M.
,
Bayna
E. M.
,
Cheresh
D. A.
(
1994
)
Receptor tyrosine kinase signaling required for integrinv 5-directed cell motility but not adhesion on vitronectin.
J. Cell Biol
127
,
859
866
Lane
E. B.
,
Hogan
B. L. M.
,
Kurkinen
M.
,
Garrels
J. I.
(
1983
)
Co-expression of vimentin and cytokeratins in parietal endoderm cells of early mouse embryo.
Nature
303
,
701
704
Leivo
I.
,
Vaheri
A.
,
Timpl
R.
,
Wartiovaara
J.
(
1980
)
Appearance and distribution of collagens and laminin in the early mouse embryo.
Dev. Biol
76
,
100
114
Letterio
J. J.
,
Geiser
A. G.
,
Kulkarni
A. B.
,
Roche
N. S.
,
Sporn
M. B.
,
Roberts
A. B.
(
1994
)
Maternal rescue of transforming growth factor-1 null mice.
Science
264
,
1936
1938
Linask
K. K.
,
Lash
J. W.
(
1988
)
A role for fibronectin in the migration of avian precardiac cells. I. Dose-dependent effects of fibronectin antibody.
Dev. Biol
129
,
315
323
Marotti
K. R.
,
Belin
D.
,
Strickland
S.
(
1982
)
The production of distinct forms of plasminogen activator by mouse embryonic cells.
Dev. Biol
90
,
154
159
Mason
I. J.
,
Taylor
A.
,
Williams
J. G.
,
Sage
H.
,
Hogan
B. L. M.
(
1986
)
Evidence from molecular cloning that SPARC, a major product of mouse embryo parietal endoderm, is related to an endothelial cell ‘culture shock’ glycoprotein of Mr43,000.
EMBO J
5
,
1465
1472
Mazariegos
M. R.
,
Leblond
C. P.
,
van der Rest
M.
(
1987
)
Radioautographic tracing of 3H-proline in the endodermal cells of the parietal yolk sac as an indicator of the biogenesis of basement membrane components.
Am. J. Anat
179
,
79
93
Nadijcka
M.
,
Hillman
N.
(
1974
)
Ultrastructural studies of the mouse blastocyst substages.
J. Embryol. Exp. Morph
32
,
675
695
Newman-Smith
E. D.
,
Werb
Z.
(
1995
)
Stem cell defects in parthenogenetic peri-implantation embryos.
Development
121
,
2069
2077
Ossowski
L.
(
1988
)
In vivo invasion of modified chorioallantoic membrane by tumor cells: the role of cell surface-bound urokinase.
J. Cell Biol
107
,
2437
2445
Rappolee
D. A.
,
Basilico
C.
,
Patel
Y.
,
Werb
Z.
(
1994
)
Expression and function of FGF-4 in peri-implantation development in mouse embryos.
Development
120
,
2259
2269
Richoux
V.
,
Darribere
T.
,
Boucaut
J. C.
,
Flechon
J. E.
,
Thiery
J. P.
(
1989
)
Distribution of fibronectins and laminin in the early pig embryo.
Anat. Rec
223
,
72
81
Sabbag
K. R.
,
Casanova
J. E.
,
Grabel
L. B.
(
1989
)
Plasminogen activator expression in F9 teratocarcinoma embryoid bodies and their endoderm derivatives.
Development
106
,
195
201
Sage
H.
,
Vernon
R. B.
,
Funk
S. E.
,
Everitt
E. A.
,
Angello
J.
(
1989
)
SPARC, a secreted protein associated with cellular proliferation, inhibits cell spreading in vitro and exhibits Ca+2-dependent binding to the extracellular matrix.
J. Cell Biol
109
,
341
356
Salamat
M.
,
Gotz
W.
,
Horster
A.
,
Janotte
B.
,
Herken
R.
(
1993
)
Ultrastructural localization of carbohydrates in Reichert's membrane of the mouse.
Cell Tissue Res
272
,
375
381
Schlafke
S.
,
Enders
A. C.
(
1963
)
Observations on the fine structure of the rat blastocyst.
J. Anat
97
,
353
360
Semoff
S.
,
Hogan
B. L. M.
,
Hopkins
C. R.
(
1982
)
Localization of fibronectin, laminin-entactin, and entactin in Reichert's membrane by immunoelectron microscopy.
EMBO J
1
,
1171
1175
Shaw
L. M.
,
Mercurio
A. M.
(
1994
)
Regulation of cellular interactions with laminin by integrin cytoplasmic domains: The A and B structural variants of the6 1 integrin differentially modulate the adhesive strength, morphology, and migration of macrophages.
Mol. Biol. Cell
5
,
679
690
Spindle
A.
(
1980
)
An improved culture medium for mouse blastocysts.
In Vitro
16
,
669
674
Spindle
A.
(
1990
)
In vitro development of one-cell embryos from outbred mice: influence of culture medium composition.
In Vitro Cell. Dev. Biol
25
,
151
156
Strickland
S.
,
Smith
K. K.
,
Marotti
K. R.
(
1980
)
Hormonal inductionof differentiation in teratocarcinoma stem cells: generation of parietal endoderm by retinoic acid and dibutyryl cAMP.
Cell
21
,
347
355
Sutherland
A. E.
,
Calarco
P. G.
,
Damsky
C. H.
(
1993
)
Developmental regulation of integrin expression at the time of implantation in the mouse embryo.
Development
119
,
1175
1186
Thorsteinsdottir
S.
(
1992
)
Basement membrane and fibronectin matrix are distinct entities in the developing mouse blastocyst.
Anat. Rec
232
,
141
149
Threadgill
D. W.
,
Dlugosz
A. A.
,
Hansen
L. A.
,
Tennenbaum
T.
,
Lichti
U.
,
Yee
D.
,
LaMantia
C.
,
Mourton
T.
,
Herrup
K.
,
Harris
R. C.
,
Barnard
J. A.
,
Yuspa
S. H.
,
Coffey
R. J.
,
Magnuson
T.
(
1995
)
Targeted disruption of mouse EGF receptor: effect of genetic background on mutant phenotype.
Science
269
,
230
234
Tienari
J.
,
Lehtonen
E.
,
Vartio
T.
,
Virtanen
I.
(
1989
)
Embryonal carcinoma cells adhere preferentially to fibronectin and laminin but their endodermal differentiation leads to a reduced adherence to laminin.
Exp. Cell Res
182
,
26
32
van de Stolpe
A.
,
Karperien
M.
,
Lowik
C. W. G. M.
,
Juppner
H.
,
Segre
G. V.
,
Abou-Samra
A. B.
,
de Laat
S. W.
,
Defize
L. H. K.
(
1993
)
Parathyroid hormone-related peptide as an endogenous inducer of parietal endoderm differentiation.
J. Cell Biol
120
,
235
243
Wartiovaara
J.
,
Leivo
I.
,
Vaheri
A.
(
1979
)
Expression of the cell surface-associated glycoprotein, fibronectin, in the early mouse embryo.
Dev. Biol
69
,
247
257
Woodley
D. T.
,
Bachmann
P. M.
,
O'Keefe
E. J.
(
1988
)
Laminin inhibits human keratinocyte migration.
J. Cell. Physiol
136
,
140
146
Wu
T.
,
Wan
Y. J.
,
Chung
A. E.
,
Damjanov
I.
(
1983
)
Immunohistochemical localization of entactin and laminin in mouse embryos and fetuses.
Dev. Biol
100
,
496
505
Yang
J. T.
,
Rayburn
H.
,
Hynes
R. O.
(
1993
)
Embryonic mesodermal defects in5integrin-deficient mice.
Development
119
,
1093
1105
Zambruno
G.
,
Marchisio
P. C.
,
Marconi
A.
,
Vaschieri
C.
,
Melchiori
A.
,
Giannetti
A.
,
De Luca
M.
(
1995
)
Transforming growth factor-1 modulates 1 and5 integrin receptors and induces the de novo expression of the v6 heterodimer in normal human keratinocytes: implications for wound healing.
J. Cell Biol
129
,
853
865
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