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Results for “Extracellular Matrix”

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PubMed1996

Effect of aprotinin on smooth muscle cell proliferation, migration, and extracellular matrix synthesis.

Aprotinin (AP) is a plasmin inhibitor commonly used to limit bleeding during reoperative coronary bypass surgery. The effect on smooth muscle cell (SMC) behavior and the development of intimal hyperplasia has not been evaluated. This study examines the effect of AP on SMC proliferation, migration, and extracellular matrix synthesis. Bovine aortic SMCs (three to five passages) plated at 5 x 10(4) cells/ml were treated with 1.0, 10, 100, and 1000 microg/ml AP. Proliferation was measured as micrograms of DNA per well, after 3 days in culture. SMC migration was measured after treatment with mitomycin C (20 microg/ml, to study migration in the absence of proliferation) by means of a fence assay. Extracellular matrix was quantitated utilizing a tritiated proline assay and corrected for cell density (cpm/microg DNA). Data are presented as means +/- SEM; ANOVA and post hoc Tuckey test were used to test for significant differences. AP stimulated SMC proliferation at dosages of 100 microg/ml (353 +/- 6 microg DNA, SMC control vs 440 +/- 11, 100 microg/ml; P < 0.05) and 1000 microg/ml (353 +/- 6, SMC control vs 503 +/- 14, 1000 microg/ml; P < 0.01). AP concentrations less than 100 microg/ml had no effect on SMC proliferation. A dose-response relationship existed between AP and SMC migration. AP doses as low as 10 microg/ml increased SMC migration from 66 +/- 4 mm2 (control SMC) to 90 +/- 4 mm2 (P < 0.02) while the maximal dose of AP (1000 microg/ml) produced an almost twofold increase in migration (66 +/- 4, SMC control vs 113 +/- 3, 1000 microg/ml group; P < 0.0001). AP had no effect on extracellular matrix synthesis. AP stimulated vascular SMC migration and proliferation but not matrix synthesis in vitro. These data suggest that AP could augment the development of intimal hyperplasia following bypass surgery.

Animals↗

PubMed1986

Migration of adult human vascular endothelial cells: effect of extracellular matrix proteins.

An ideal vascular prosthetic graft should support rapid endothelial coverage and allow maximal rates of endothelial cell migration on its surface. To study this we modified and developed a method to measure rates of migration of adult human vascular endothelial cells (AHVECs) on test surfaces using an O-ring fence technique. We measured the rates of AHVEC migration on a variety of extracellular matrix proteins of interest because of their ability to support cell attachment. AHVECs, initially constrained in rings with an area of 0.62 cm2 (day 0), were allowed to migrate radially, and area coverage was measured after 7 days. The greatest areas of migration were seen on fibronectin (1.20 cm2) and gelatin (1.29 cm2) when compared with control (polystyrene [0.97 cm2]), p less than 0.01, p less than 0.05. Migration on laminin and prepared extracellular matrix from AHVEC was not statistically different from that of controls. Of extracellular matrix proteins studied in our system, a gelatin substrate in the presence of serum resulted in maximal rates of AHVEC migration.

Blood Vessel Prosthesis↗

PubMed2003

Matrix metalloproteinase activity in urine of patients with renal cell carcinoma leads to degradation of extracellular matrix proteins: possible use as a screening assay.

PURPOSE: Localized renal cell carcinoma is usually curable by nephrectomy. However, a large fraction of patients already present with metastatic disease, which results in a poor outcome. Currently no clinically relevant screening assay is available to detect early stage renal cell carcinoma. We investigated whether urinary extracellular matrix (ECM) proteins and/or matrix metalloproteinase (MMP) activity may be valuable as a noninvasive indicator of early stage renal cell carcinoma. MATERIALS AND METHODS: Urine specimens from preoperative patients with renal cell carcinoma and healthy controls were collected. The urinary excretion of the ECM proteins collagen IV, laminin and fibronectin was investigated by immunoblotting. MMP activity was assessed by gelatin zymography and by a fluorescence based microtiter plate activity assay. RESULTS: The full-length forms of all 3 ECM proteins investigated were significantly decreased or absent in renal cell carcinoma urine. Based on criteria established in this study this finding would lead to the correct detection of 95% of patients with renal cell carcinoma (21 of 22) with a false-positive rate of 4.5% (1 of 22 controls). All 11 nonmetastatic cases of the lowest clinical stage (T1N0M0) were correctly identified. The absence of urinary ECM proteins was due to significantly increased urinary MMP activity. CONCLUSIONS: Analysis of decreased urinary ECM proteins and analysis of increased MMP activity may have value for the development of a sensitive, high throughput molecular screening assay to detect early stage renal cell carcinoma.

Adult↗

PubMed2001

Increased extracellular matrix remodeling is associated with tumor progression in human hepatocellular carcinomas.

Matrix metalloproteinase-2 (MMP2) is a key enzyme in the process of extracellular matrix remodeling involved in tumor invasion and metastasis. The activation of MMP2 involves interplay with the membrane type-matrix metalloproteinase-1 (MT1-MMP) and the tissue inhibitor of metalloproteinase-2 (TIMP2). In vitro, activated hepatic stellate cells are a main source of MMP2 and collagen I induces MMP2 activation. The steady-state mRNA levels of MMP2, MT1-MMP, TIMP2, collagen I, collagen IV, and laminin gamma1 were compared with MMP2 activity in 55 hepatocellular carcinomas, 47 matching nontumor biopsies and 19 histologically normal livers. In hepatocellular carcinomas, increased collagen I mRNA levels were strongly associated with those of MMP2 (Spearman R =.74, P <.001), MT1-MMP (R =.65, P <.001) and TIMP2 (R = 0.61, P <.001). MMP2 activity was correlated with the mRNA expression of collagen I (R =.45 P <.01), collagen IV (R =.40, P <.01) and laminin gamma1 (R =.33, P <.05). Unlike collagen IV and laminin gamma1 mRNAs, MMP2, MT1-MMP, TIMP2, collagen I mRNA levels were increased in nonencapsulated compared with encapsulated tumors (P <.05). In addition, MMP2 activity was fourfold higher (P <.01) in tumors arising in cirrhotic livers than in those arising in noncirrhotic livers. Moreover, tumor recurrence was associated with 4.6- and 2.8-fold (P <.05) higher collagen I and MMP2 mRNA levels, respectively, in hepatocellular carcinomas arising in cirrhotic livers. Thus, a high extracellular matrix remodeling favors tumor progression in hepatocellular carcinomas.

Aged↗

PubMed1996

Mouse polymorphonuclear granulocyte binding to extracellular matrix molecules involves beta 1 integrins.

The mechanism of adhesion of purified mouse polymorphonuclear granulocytes (PMN) to extracellular matrix proteins characteristic of basement membranes and the interstitium has been investigated and compared with the adhesion of a mouse progranulocytic cell line, 32DC13, and a mouse monocytic cell line, WEHI 78/24. All three cell types bound specifically to fibronectin and vitronectin to different degrees under different cellular activation states. 32DC13 bound to fibronectin and vitronectin strongly, and this binding increased upon cellular activation with phorbol 12-myristate-13-acetate (PMA) but not with formyl-Met-Leu-Phe. Only 32DC13 showed significant binding to laminin-1. By contrast, WEHI 78/24 and PMN bound only fibronectin and vitronectin; this binding was weak and was altered only marginally upon activation with PMA. In the case of WEHI 78/24, a slight increase in adhesion both to fibronectin and to vitronectin was observed after cellular activation with PMA, while PMN adhesion to both substrates was slightly reduced. The mechanism of binding to fibronectin and vitronectin was similar in the three cell types. The integrin alpha5 beta1 mediated fibronectin adhesion, demonstrating for the first time the existence of a functionally active beta1 integrin on mouse PMN. Vitronectin binding was mediated by alpha(v) beta3, as demonstrated by the ability of alpha(v)-specific cyclic L-Arg-L-Gly-L-Asp-D-Phe-L-Val (RGDfV) peptide (EMD66203), and anti-beta3 antibody to inhibit cell adhesion. 32DC13 adhesion to laminin-1 was via the alpha6 beta1 integrin. None of the three cell types tested bound to the basement membrane proteins collagen type IV and perlecan, or to the interstitial stromal constituents tenascin, collagen types I, V and VI. Interestingly, perlecan and collagen type IV were found to repel all three cell types. The relative inability of PMN, WEHI 78/24, and 32DC13 to bind to extracellular matrix proteins characteristic of basement membranes and their ability to bind inflammatory markers of the interstitium is discussed with respect to leukocyte extravasation processes.

Animals↗

PubMed1995

Focal nodular hyperplasia of the liver: composition of the extracellular matrix and expression of cell-cell and cell-matrix adhesion molecules.

We studied by immunohistochemistry 25 cases of focal nodular hyperplasia (FNH) to evaluate the composition of the extracellular matrix and the expression and distribution of endothelial cell-cell adhesion molecules and integrin receptors. The extracellular matrix of FNH retained the overall organization of that of normal liver. The matrix of central scars resembled that of portal tracts. The main difference was the presence of large vitronectin deposits, which might indicate the existence of local hemodynamic disturbances. The matrix lining the sinusoid-like vessels running in the hyperplastic parenchyma retained characteristic features of the normal perisinusoidal matrix, such as the presence of tenascin. In the zone surrounding the central scars, it contained large amounts of laminin, von Willebrand factor, and thrombospondin, suggesting the development of perisinusoidal fibrosis. Laminin deposition was accompanied by the induction of cell-cell adhesion molecules on adjacent endothelial cells and by the up-regulation of specific integrin receptors on both hepatocytes and sinusoidal endothelial cells. In conclusion, our study: (1) reinforces the hypothesis that FNH is merely a hyperplastic response of liver parenchyma to local vascular abnormalities, and (2) shows that the lesions of perisinusoidal fibrosis associated with FNH are accompanied by the induction of integrin receptors on hepatocytes and sinusoidal endothelial cells.

Adult↗

PubMed2004

[Extracellular matrix for the replacement of ureteral defect].

This is an experiment on rabbits to evaluate the possibility of ureteral replacement by extracellular matrix. We adopted a biochemical method for preparing a new tissue engineering material named Extracellular Matrix (ECM), and the ECMs were used as homologous grafts to replace the defect in the ureters. Light microscopy, scanning electron microscopy, immunohistochemical technique and intravenous urography were used. The routine blood and biochemical laboratory tests were made before and after operation, and the measured values of pressure in the ureter of experiment and control groups were compared. The ureteral ECM was found in the experiment to promote the regeneration of all ureteral wall components. There were no significant differentces between the regenerative tissue and the normal tissue in morphology and function 16 weeks after replacement. The homologous ECM might be an ideal replacement material for ureteral defect.

Animals↗

PubMed1994

Differences in binding to the solid substratum and extracellular matrix may explain isoform-specific paracrine effects of platelet-derived growth factor.

We have studied the paracrine response of fibroblasts to the two homodimeric isoforms of platelet-derived growth factor (PDGF-AA and -BB). CHO-cells stably transfected with a B-chain cDNA expression vector (CHO-PDGF-B cells), were found to elicit a marked paracrine response when seeded at clonal density on preformed monolayers of human or murine fibroblasts; no such response was elicited by CHO-PDGF-A cells. Immunofluorescence microscopy of CHO-PDGF-B cell cultures showed the presence of a pericellular deposit of material reacting with antibodies against PDGF-BB; no corresponding PDGF-AA immunoreactive material was found in the CHO-PDGF-A cultures. The pericellular material, deposited by CHO-PDGF-B cells, was shown to have a growth promoting effect on target cells. Furthermore, we could show that 125I-PDGF-BB binds more efficiently than 125I-PDGF-AA to extracellular matrix prepared from foreskin fibroblast cultures, as well as to defined extracellular matrix components (fibronectin, laminin and collagen III). The results reveal a marked difference in the paracrine activity of PDGF-AA and PDGF-BB; the latter has a strong local growth enhancing effect, that is mostly likely to be ascribed to its association with components of the extracellular matrix.

3T3 Cells↗

PubMed1987

Ultrastructural tissue morphometry of the distribution of extracellular matrix vesicles in remodeling rat tibial bone six days after injury.

A study of the distribution of extracellular matrix vesicles on the 6th day of bone healing was performed by methods of transmission electron microscopy combined with computerized morphometry. The vesicles were detected on the electron micrographs and grouped according to their diameters, distance from the calcified front and type. The different types were selected as follows: vesicles with electron-lucent contents, i.e., 'empty'; vesicles with amorphous electron-opaque contents, i.e., 'amorphous'; vesicles containing crystalline depositions, i.e., 'crystal', and vesicles containing crystalline structures with ruptured membranes, i.e., 'rupture'. Most of the vesicles were concentrated between diameters of 0.02 and 0.22 microns. Most of the vesicles were found within a distance of less than 3 micron from the calcified front. The vesicles were distributed according to their types: 'empty', 'amorphous', 'crystal' and 'rupture' in 14, 39, 34 and 13%, respectively. The diameters of the 'crystal' and 'rupture' vesicles were significantly larger than those of the 'empty' and 'amorphous' types. The sequence of distances from the calcified front was recorded as follows: 'rupture', 'crystal', 'amorphous' and 'empty', the 'rupture' type being the closest to the front. The results of the present study confirm the accepted hypothesis on calcification via extracellular matrix vesicles. It is thought that the cell secretes 'empty' vesicles that accumulate amorphous Ca and P to form a hydroxy-apatite crystal. This is followed by rupture of the vesicular membrane. The propagation of the process is accompanied by increase in the vesicular diameter and its approximation to the calcifying front.

Animals↗

PubMed1996

The influence of extracellular matrix substrata on preadipocyte development in serum-free cultures of stromal-vascular cells.

The influence of the extracellular matrix (ECM) and ECM components on preadipocyte development was examined in primary cultures of adipose tissue stromal-vascular (S-V) cells. Extracellular matrix derived from Engelbreth-Holm-Swarm (EHS) cells or tumors enhanced several aspects of adipogenesis in vitro. In comparison to uncoated and fibronectin substrata, EHS-ECM substratum markedly increased attachment, spreading, and hypertrophy of preadipocytes while antagonizing spreading of non-preadipocytes. In addition, adipocyte number increased (P < .05) on these substrata despite no increase in total cell number: this resulted in a greater (P < .05) proportion of preadipocytes. These effects of EHS-ECM were also observed with laminin substrata per se, whereas types I and IV collagen and fibronectin had no influence. In contrast to all other substrata, adipocyte number decreased and total cell number increased 2.5-fold on ECM derived from corneal endothelial cells; this resulted in the lowest proportion of preadipocytes. Challenging cultures with adipogenic media (+serum) did not counter the inhibitory influence of corneal endothelial ECM, whereas dexamethasone partially neutralized the inhibitory influence of this ECM. These studies clearly show that source or type of the ECM dictated the influence of ECM substrata on preadipocyte development in primary S-V cultures. However, these studies indicated that the ECM and in particular laminin may play a critical role in morphological aspects of preadipocyte development.

Adipocytes↗

PubMed1995

Extracellular matrix proteins and basement membrane: their identification in bovine ovaries and significance for the attachment of cultured preantral follicles.

Described in the present paper is the immunolocalization of the extracellular matrix proteins (e.g., fibronectin, collagen Types I and III) in the bovine ovary, with special attention to preantral follicles. In addition, we have shown, histochemically and ultrastructurally, that mechanically isolated bovine preantral follicles are surrounded by an intact basement membrane. After 24 h of culture in serum-free medium, only 20.4% of these follicles attached to a plastic substrate. We showed that covering the plastic with extracellular matrix proteins (i.e., fibronectin, collagen Type I and matrigel) significantly increased the percentage of attached follicles to 76.0, 65.2 and 80.4%, respectively, while laminin had no effect (18.6%). When preantral follicles were embedded within three-dimensional collagen gels, no loss of follicles was observed. Restoring surface interactions between preantral follicles and the extracellular matrix in vitro, either in a two- or a three-dimensional system, might be important for maintaining follicular viability and growth in the future.

Journal Article↗

PubMed1998

Fibronectin and the extracellular matrix in the perforating disorders of the skin.

Despite detailed microscopic descriptions and clinical observation, little is known regarding the pathogenesis of the perforating disorders of skin, which have traditionally been subdivided into numerous microscopic entities associated with various clinical settings. An increasing body of evidence now suggests that the perforating disorders of skin are akin, and may constitute an expanded single pathologic entity. Each of the classic perforating disorders of skin, including elastosis perforans serpiginosa, perforating folliculitis, reactive perforating collagenosis, Kyrle's disease, and perforating disorder of uremia, have been shown to extrude collagen, elastin, and related extracellular matrix components through the epidermis. Considering a shared pathogenic mechanism among these entities, we explored the possible role of the extracellular matrix, in particular fibronectin, in perforating disorders of skin. Using immunohistochemical and serum determinations of extracellular matrix constituents, including fibronectin, collagen type IV, laminin, and tenascin, we showed consistent serum elevation and/or deposition of fibronectin, in each case, without a commensurate increase in laminin, collagen type IV, and tenascin. We propose that elevated serum and tissue concentrations of fibronectin may be responsible for inciting, in a physiologically aberrant manner, increased epithelial migration and proliferation culminating in perforation.

Adolescent↗

PubMed2002

Cancer as a disease of epithelial-mesenchymal interactions and extracellular matrix regulation.

Carcinogenesis - the process of cancer formation - is commonly discussed in terms of genetic alterations that lead to deregulation of cell growth. Recently, there has been a resurgence of interest in epigenetic factors and, in particular, the role of the stromal microenvironment and angiogenesis in tumor formation. In this article, cancer is presented as a disease of the developmental processes that govern how cells organize into tissues and tissues into organs. This histogenetic perspective raises the possibility that epithelial-mesenchymal interactions and the extracellular matrix (basement membrane) that is deposited through these interactions may actively contribute to the carcinogenic process. Experimental work is reviewed that confirms that extracellular matrix plays a key role in normal histodifferentiation during both epitheliogenesis and angiogenesis, and that epigenetic deregulation of cell-matrix interactions may actively promote tumor initiation and progression. The contributions of integrins, cytoskeleton, tensegrity and local variations in extracellular matrix mechanics to these processes are discussed, as are the implications of this work for future studies on cancer formation.

Animals↗

PubMed1994

Effects of cytokines on gingival fibroblasts in vitro are modulated by the extracellular matrix.

Fibroblast function in gingival tissue is thought to be regulated by the local cellular environment--both the extracellular matrix and soluble factors. In an attempt to artificially re-create this situation fibroblasts have been cultured within 3-dimensional collagen gels in an environment more physiologically comparable to connective tissue. Using such a model we investigated the effects of the extracellular matrix on gingival fibroblast growth and synthetic activity and on the cellular responsiveness to 4 soluble factors--epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factor-beta (TGF-beta 1) and interleukin-1 beta (IL-1 beta). Fibroblasts cultured within collagen gels showed similar growth rates, an increased production of collagen but reduced levels of hyaluronan synthesis in comparison to cells in monolayer culture. Cellular responsiveness to soluble mediators was also modulated by the collagen matrix, with a generalised reduction in response by cells embedded within the matrix. The stimulatory effects of EGF and PDGF on cell growth in monolayer over a 14-day period were only found during the initial stages of culture within gels. Similarly the stimulation of matrix production by cells induced by TGF-beta 1 on plastic was reduced or even negated when cells were cultured in collagen gels. On plastic IL-1 beta significantly stimulated cell growth but had no effect on either collagen or hyaluronan production by fibroblasts. In gel cultures, this cytokine had no effect on cell proliferation, but significantly inhibited both collagen and hyaluronan synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1998

Effects of extremely low frequency electromagnetic field (EMF) on collagen type I mRNA expression and extracellular matrix synthesis of human osteoblastic cells.

Human osteoblastic cells were grown in a three-dimensional (3-D) cell culture model and used to test the effects of a 20 Hz sinusoidal electromagnetic field (EMF; 6 mT and 113 mV/cm max) on collagen type I mRNA expression and extracellular matrix formation in comparison with the effects of growth factors. The cells were isolated from trabecular bone of a healthy individual (HO-197) and from a patient presenting with myositis ossificans (MO-192) and grown in a collagenous sponge-like substrate. Maximal enhancement of collagen type I expression after EMF treatment was 3.7-fold in HO-197 cells and 5.4-fold in MO-192 cells. Similar enhancement was found after transforming growth factor-beta (TGF-beta) and insulin-like growth factor-I (IGF-I) treatment. Combined treatment of the cells with EMF and the two growth factors TGF-beta and IGF-I did not act synergistically. MO-192 cells produced an osteoblast-characteristic extracellular matrix containing collagen type I, alkaline phosphatase, and osteocalcin, together with collagen type III, TP-1, and TP-3, two epitopes of an osteoblastic differentiation marker. The data suggest that the effects of EMFs on osteoblastic differentiation are comparable to those of TGF-beta and IGF-I. We conclude that EMF effects in the treatment of skeletal disorders and in orthopedic adjuvant therapy are mediated via enhancement of collagen type I mRNA expression, which may lead to extensive extracellular matrix synthesis.

Cell Line↗

PubMed1996

Effects of hypoxia on the extracellular matrix production of cultured rat mesangial cells.

Glomerular changes in patients with cyanotic congenital heart and chronic lung diseases and in persons living at a high altitude might be related to hypoxemia. This study was carried out to examine the effects of hypoxia on the extracellular matrix production by cultured rat mesangial cells (CRMC). Subconfluent CRMC monolayers were grown with 10% oxygen (hypoxia) and 20% oxygen (control) for 1, 3, and 5 days. The production of type IV collagen (CIV), fibronectin (FN), and laminin (LN) by CRMC was evaluated by flow cytometry and immunofluorescent microscopy. Total RNA was extracted and Northern blotting and hybridization were performed with cDNAs for CIV, FN, and LN. The surface expression of CIV on FC was higher in hypoxia than under control conditions at day 5 (158% of control). The surface expression of FN was also higher in hypoxia at day 3 (303%) and at day 5 (332%). The surface expression of LN was lower at day 1 (71%). Immunofluorescent microscopy showed similar changes with flow cytometry. The mRNA level for CIV and FN was maximal at day 5 with 206 and 305% of control, respectively. Hypoxia had little effect on LN mRNA expression. These results show that hypoxia stimulates the synthesis of extracellular matrix of cultured rat mesangial cells. Hypoxia may contribute to the development of glomerular changes in cyanotic congenital heart diseases, chronic lung diseases, and in persons living at a high altitude.

Animals↗

PubMed2001

Early activation of hepatic stellate cells and perisinusoidal extracellular matrix changes during ex vivo pig liver perfusion.

In previous works, we observed during liver transplantation procedure, the early activation of hepatic stellate cells (HSC) which acquire alpha-smooth muscle (SM) actin expression. In this study, we evaluated changes in HSC and in perisinusoidal extracellular matrix during ex vivo pig liver perfusion. Under general anesthesia, pig livers were flushed and removed, and then perfused ex vivo for 6 h with homologous blood. Liver biopsies were taken before and after washout, at 5 min perfusion, and then hourly. Tissues were processed for immunohistochemistry, immunofluorescence, confocal microscopy, in situ hybridization and electron microscopy. Before and after liver washout, alpha-SM actin was present in vessel walls but in very few lobular HSC. After 1 h perfusion, a strong reactivity for alpha-SM actin was present in HSC, particularly along dilated sinusoids. At the ultrastructural level, numerous microfilament bundles appeared in HSC cytoplasmic processes. During perfusion, type I and type IV collagens, type III procollagen, and fibronectin acquired a looser organisation in relation with the enlargement of perisinusoidal spaces; laminin appeared in perisinusoidal spaces around portal areas and fibrillin deposits increased. In situ hybridization studies showed an increase of the type I procollagen mRNA expression mainly in portal tracts and septa. Ex vivo liver perfusion induces: 1) an early activation of HSC which acquire the expression of alpha-SM actin, and 2) significant changes in the perisinusoidal extracellular matrix. These results are compatible with the view that HSC function as liver specific pericytes participating in the regulation of sinusoidal blood pressure.

Actin Cytoskeleton↗

PubMed1988

Expression and function of cell surface extracellular matrix receptors in mouse blastocyst attachment and outgrowth.

Mouse-hatched blastocysts cultured in vitro will attach and form outgrowths of trophoblast cells on appropriate substrates, providing a model for implantation. Immediately after hatching, the surfaces of blastocysts are quiescent and are not adhesive. Over the period 24-36 h post-hatching, blastocysts cultured in serum-free medium become adhesive and attach and spread on the extracellular matrix components fibronectin, laminin, and collagen type IV in a ligand specific manner. Attachment and trophoblast outgrowth on these substrates can be inhibited by addition to the culture medium of an antibody, anti-ECMr (anti-extracellular matrix receptor), that recognizes a group of 140-kD glycoproteins similar to those of the 140-kD extracellular matrix receptor complex (integrin) recognized in avian cells by CSAT and JG22 monoclonal antibodies. Addition to the culture medium of a synthetic peptide containing the Arg-Gly-Asp tripeptide cell recognition sequence of fibronectin inhibits trophoblast outgrowth on both laminin and fibronectin. However, the presence of the peptide does not affect attachment of the blastocysts to either ligand. Immunoprecipitation of 125I surface-labeled embryos using anti-ECMr reveals that antigens recognized by this antibody are exposed on the surfaces of embryos at a time when they are spreading on the substrate, but are not detectable immediately after hatching. Immunofluorescence experiments show that both the ECMr antigens and the cytoskeletal proteins vinculin and talin are enriched on the cell processes and ventral surfaces of trophectoderm cells in embryo outgrowths, in patterns similar to those seen in fibroblasts, and consistent with their role in adhesion of the trophoblast cells to the substratum.

Animals↗