The trophectoderm layer of the mouse blastocyst differentiates at the late blastocyst stage to form the invasive trophoblast that mediates implantation of the embryo into the uterine wall. The first sign that trophoblast cells have developed an invasion-specific cell behavior appears about 10–15 hours after the embryo hatches from the zona pellucida, when the quiescent, non-adherent trophectoderm cells initiate protrusive activity and become adhesive to extracellular matrix. Our previous findings that trophoblast outgrowth on extracellular-matrix-coated substrata involves the integrin family of adhesion receptors (Sutherland, A. E., Calarco, P. G. and Damsky, C. H., 1988, J. Cell Biol. 106, 1331–1348), suggested that the onset of trophoblast adhesive and migratory behavior at the time of implantation may be due to changes in expression or distribution of integrin receptors. We have thus examined the mRNA and protein expression of individual integrin subunits during pre- and periimplantation development (E0-E7.5). A basic repertoire of integrins, including receptors for fibronectin (alpha 5 beta 1), laminin (alpha 6B beta 1) and vitronectin (alpha v beta 3), was expressed continuously throughout this period, whereas the expression of five other integrin subunits was developmentally regulated. The mRNA for three of these (alpha 2, alpha 6A and alpha 7) was first detected in the late blastocyst, coincident with endoderm differentiation and development of attachment competence. The mRNA for another (alpha 1) was not detected until after trophoblast outgrowth had begun, suggesting that its expression may be induced by contact with matrix. At E7.5, three of the temporally regulated integrins (alpha 1, apha 6A, alpha 7), all of which can form receptors for laminin, were detected only in the ectoplacental cone (differentiating trophoblast), and may thus play specific roles in trophoblast adhesion and/or differentiation. Because laminin expression is upregulated in decidualized uterine stroma in response to the implanting embryo, we examined trophoblast-laminin interactions, using laminin fragments and integrin antibodies to determine which integrin receptors were involved. Trophoblast cells attached and spread on both the E8 and P1′ fragments of laminin; however, the P1′ binding site was cryptic in intact laminin. Interaction with P1′ was RGD- and alpha v beta 3-dependent, whereas outgrowth on E8 was RGD-independent and not inhibited by antibodies to the laminin receptor alpha 6 beta 1, suggesting that alpha 7 beta 1 is the major trophoblast integrin E8 receptor.(ABSTRACT TRUNCATED AT 400 WORDS)

Armant
D. R.
(
1991
).
Cell interactions with laminin and its proteolytic fragments during outgrowth of mouse primary trophoblast cells.
Biol. Reprod
45
,
664
672
Armant
D. R.
,
Kaplan
H. A.
,
Mover
H.
,
Lennarz
W. I.
(
1986
).
The effect of hexapeptides on attachment and outgrowth of mouse blastocysts cultured in vitro: Evidence for the involvement of the cell recognition tripeptide Arg-Gly-Asp.
Proc. Natl. Acad. Sci. USA
83
,
6751
6755
Aumailley
M.
,
Gurrath
M.
,
Muller
G.
,
Calvete
J.
,
Timpl
R.
,
Kessler
H.
(
1991
).
Arg-Gly-Asp constrained within cyclic pentapeptides. Strong and selective inhibitors of cell adhesion to vitronectin and laminin fragment P1.
FEBS Letters
291
,
50
54
Bayna
E. M.
,
Shaper
J. H.
,
Shur
B. D.
(
1988
).
Temporally specific involvement of cell surface beta-1,4 galactosyltransferase during mouse embryo morula compaction.
Cell
53
,
145
157
Behrendtsen
O.
,
Alexander
C. M.
,
Werb
Z.
(
1992
).
Metalloproteinases mediate extracellular matrix degradation by cells from mouse blastocyst outgrowths.
Development
114
,
447
456
Bevilacqua
E. M. A. F.
,
Abrahamsohn
P. A.
(
1988
).
Ultrastructure of trophoblast giant cell transformation during the invasive stage of implantation of the mouse embryo.
J. Morphol
198
,
341
351
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
Clegg
K. B.
,
Piko
L.
(
1983
).
Quantitative aspects of RNA synthesis and polyadenylation in 1-cell and 2-cell mouse embryos.
J. Embryol. Exp. Morph
74
,
169
182
Cooper
A. R.
,
MacQueen
H. A.
(
1983
).
Subunits of laminin are differentially synthesized in mouse eggs and early embryos.
Dev. Biol
96
,
467
471
Cooper
H. M.
,
Tamura
R. N.
,
Quaranta
V.
(
1991
).
The major1186laminin receptor of mouse embryonic stem cells is a novel isoform of the 61 integrin.
J. Cell Biol
115
,
843
850
Damsky
C. H.
,
Fitzgerald
M. L.
,
Fisher
S. J.
(
1992
).
Distribution patterns of extracellular matrix components and adhesion receptors are intricately modulated during first trimester cytotrophoblast differentiation along the invasive pathway, in vivo.
J. Clin. Invest
89
,
210
222
Damsky
C. H.
,
Richa
J.
,
Solter
D.
,
Knudsen
K.
,
Buck
C. A.
(
1983
).
Identification and purification of a cell-surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue.
Cell
34
,
455
466
Flach
G.
,
Johnson
M. H.
,
Braude
P. R.
,
Taylor
R. A. S.
,
Bolton
V. N.
(
1982
).
The transition from maternal to embryonic control in the 2-cell mouse embryo.
EMBO J
1
,
681
686
Forsberg
E.
,
Paulsson
M.
,
Timpl
R.
,
Johansson
S.
(
1990
).
Characterization of a laminin receptor on rat hepatocytes.
J. Biol. Chem
265
,
6376
6381
Gehlsen
K. R.
,
Sriramarao
P.
,
Furcht
L. T.
,
Skubitz
A. P.
(
1992
).
A synthetic peptide from the carboxy terminus of the laminin A chain represents a binding site for the alpha 3 beta 1 integrin.
J. Cell Biol
117
,
449
459
Giebelhaus
D. H.
,
Heikkila
J. J.
,
Schultz
G. A.
(
1983
).
Changes in the quantity of histone and actin messenger RNA during the development of preimplantation mouse embryos.
Dev. Biol
98
,
148
154
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
Glasser
S. R.
,
Lampelo
L.
,
Munir
M. I.
,
Julian
J.
(
1987
).
Expression of desmin, laminin and fibronectin during in situ differentiation (decidualization) of rat uterine stromal cells.
Differentiation
35
,
132
142
Goodman
S. L.
,
Aumailley
M.
,
von der Mark
H.
(
1991
).
Multiple cell surface receptors for the short arms of laminin: alpha 1 beta 1 integrin and RGD-dependent proteins mediate cell attachment only to domains III in murine tumor laminin.
J. Cell Biol
113
,
931
941
Hall
D. E.
,
Reichardt
L. F.
,
Crowley
E.
,
Holley
B.
,
Moezzi
H.
,
Sonnenberg
A.
,
Damsky
C. H.
(
1990
).
The1 /1 and6 /1 integrin heterodimers mediate cell attachment to distinct sites on laminin.
J. Cell Biol
110
,
2175
2184
Hogervorst
F.
,
Kuikman
I.
,
van Kessel
A. G.
,
Sonnenberg
A.
(
1991
).
Molecular cloning of the human alpha 6 integrin subunit. Alternative splicing of alpha 6 mRNA and chromosomal localization of the alpha 6 and beta 4 genes.
Eur. J. Biochem
199
,
425
433
Holers
V. M.
,
Ruff
T. G.
,
Parks
D. L.
,
McDonald
J. A.
,
Ballard
L. L.
,
Brown
E. J.
(
1989
).
Molecular cloning of a murine fibronectin receptor and its expression during inflammation.
J. Exp. Med
169
,
1589
1605
Ignatius
M. A.
,
Large
T. H.
,
Houde
M.
,
Tawil
J. W.
,
Barton
A.
,
Esch
F.
,
Carbonetto
S.
,
Reichardt
L.
(
1990
).
Molecular cloning of the rat integrin1 -subunit: A receptor for laminin and collagen.
J. Cell Biol
111
,
709
720
Jenkinson
E. J.
,
Wilson
I. B.
(
1973
).
In vitro studies on the control of trophoblast outgrowth in the mouse.
J. Embryol.Exp. Morph
30
,
21
30
Johnson
M. H.
,
Rossant
J.
(
1981
).
Molecular studies on cells of the trophectodermal lineage of the postimplantation mouse embryo.
J. Embryol. Exp. Morph
61
,
103
116
Kramer
R. H.
,
Vu
M. P.
,
Cheng
Y. F.
,
Ramos
D. M.
,
Timpl
R.
,
Waleh
N.
(
1991
).
Laminin-binding integrin7 1: functional characterization and expression in normal and malignant melanocytes.
Cell Regulation
2
,
805
817
Librach
C.
,
Fitzgerald
M.
,
Fisher
S. J.
,
Damsky
C. H.
(
1991
).
Cytotrophoblast-fibronectin and cytotrophoblast-laminin interactions have distinct roles in cytotrophoblast invasion. (Abstract).
J. Cell Biol
115
,
6
–.
Mecham
R. P.
(
1991
).
Laminin receptors.
Annu. Rev. Cell Biol
7
,
71
91
Nurcombe
V.
,
Aumailley
M.
,
Timpl
R.
,
Edgar
D.
(
1989
).
The high-affinity binding of laminin to cells. Assignation of a major cell-binding site to the long arm of laminin and of a latent cell-binding site to its short arms.
Eur. J. Biochem
180
,
9
14
O'Shea
K. S.
,
Liu
L. H.
,
Kinnunen
L. H.
,
Dixit
V. M.
(
1990
).
Role of the extracellular matrix protein thrombospondin in the early development of the mouse embryo.
J. Cell Biol
111
,
2713
2723
Rappolee
D. A.
,
Mark
D.
,
Banda
M. J.
,
Werb
Z.
(
1988
).
Woundmacrophages express TGF-alpha and other growth factors in vivo: analysis by RNA phenotyping.
Science
241
,
708
712
Richa
J.
,
Damsky
C. H.
,
Buck
C. A.
,
Knowles
B. B.
,
Solter
D.
(
1985
).
Cell surface glycoproteins mediate compaction, trophoblast attachment, and endoderm formation during early mouse development.
Dev. Biol
108
,
513
521
Romagnano
L.
,
Babiarz
B.
(
1990
).
The role of murine cell surface galactosyltransferase in trophoblast-laminin interactions in vitro.
Dev. Biol
141
,
254
261
Rossino
P.
,
Gavazzi
I.
,
Timpl
R.
,
Aumailley
M.
,
Abbadini
M.
,
Giancotti
F.
,
Silengo
L.
,
Marchiso
P. C.
,
Tarone
G.
(
1990
).
Nerve growth factor induces increased expression of a laminin-binding integrin in rat pheochromocytoma PC12 cells.
Exp. Cell Res
189
,
100
108
Sang
Q.-X.
,
Thompson
E. W.
,
Grant
D.
,
Stetler-Stevenson
W. G.
,
Byers
S. W.
(
1991
).
Soluble laminin and arginine-glycine-aspartic acid containing peptides differentially regulate type IV collagenase messenger RNA, activation, and localization in testicular cell culture.
Biol. Reprod
45
,
387
394
Schlafke
S.
,
Enders
A. C.
(
1975
).
Cellular basis of interaction between trophoblast and uterus at implantation.
Biol. Reprod
12
,
41
65
Senior
P. V.
,
Critchley
E. R.
,
Beck
F.
,
Walker
R. A.
,
Varley
J. M.
(
1988
).
The localization of laminin mRNA and protein in the postimplantation embryo and placenta of the mouse: An in situ hybridization and immunocytochemical study.
Development
104
,
431
446
Spindle
A.
(
1980
).
An improved culture medium for mouse blastocysts.
In Vitro
16
,
669
674
Spindle
A.
,
Pederson
R. A.
(
1973
).
Hatching, attachment, and outgrowth of mouse blastocysts in vitro: fixed nitrogen requirements.
J. Exp. Zool
186
,
305
318
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 that 61 but not6 4functions as a major receptor for fragment E8.
J. Cell Biol
110
,
2145
2155
Sutherland
A. E.
,
Calarco
P. G.
,
Damsky
C. H.
(
1988
).
Expression and function of cell surface extracellular matrix receptors in mouse blastocyst attachment and outgrowth.
J. Cell Biol
106
,
1331
1348
Sutherland
A. E.
,
Sanderson
R. D.
,
Mayes
M.
,
Siebert
M.
,
Calarco
P. G.
,
Bernfield
M.
,
Damsky
C. H.
(
1991
).
Expression of syndecan, a putative low affinity fibroblast growth factor receptor, in the early mouse embryo.
Development
113
,
339
351
Takada
Y.
,
Hemler
M. E.
(
1989
).
The primary structure of the VLA-2/collagen receptor2 subunit (platelet GPIa): homology to other integrins and the presence of a possible collagen-binding domain.
J. Cell Biol
109
,
397
407
Tamura
R. N.
,
Cooper
H. M.
,
Collo
G.
,
Quaranta
V.
(
1991
).
Cell type-specific integrin variants with alternativechain cytoplasmic domains.
Proc. Natl. Acad. Sci. USA
88
,
10183
10187
Terranova
V. P.
,
Williams
J. E.
,
Liotta
L. A.
,
Martin
G. R.
(
1984
).
Modulation of the metastatic activity of melanoma cells by laminin and fibronectin.
Science
226
,
982
985
Thorsteinsdottir
S.
(
1992
).
Basement membrane and fibronectin matrix are distinct entities in the developing mouse blastocyst.
Anat. Rec
232
,
141
149
Tsuji
T.
,
Yamamoto
F.
,
Miura
Y.
,
Takio
K.
,
Titani
K.
,
Pawar
S.
,
Osawa
T.
,
Hakomori
S.
(
1990
).
Characterization through cDNA cloning of galactoprotein b3 (Gapb3), a cell surface membrane glycoprotein showing enhanced expression on oncogenic transformation. Identification of Gapb3 as a member of the integrin superfamily.
J. Biol. Chem
265
,
7016
7021
Turpeenniemi-Hujanen
T.
,
Thorgeirsson
U. P.
,
Rao
C. N.
,
Liotta
L. A.
(
1986
).
Laminin increases the release of type IV collagenase from malignant cells.
J. Biol. Chem
261
,
1883
1889
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
Wewer
U. M.
,
Damjanov
A.
,
Weiss
J.
,
Liotta
L. A.
,
Damjanov
I.
(
1986
).
Mouse endometrial stromal cells produce basement-membrane components.
Differentiation
32
,
49
58
Yurchenko
P. D.
,
Schittny
J. C.
(
1990
).
Molecular architecture of basement membranes.
FASEB J
4
,
1577
1590
This content is only available via PDF.