ARTICLE |
Correspondence to: Lari Häkkinen, U. of British Columbia, Faculty of Dentistry, Dept. of Oral Biological and Medical Sciences, 2199 Wesbrook Mall, Vancouver, BC, Canada V6T 1Z3. E-mail: lhakkine@interchange.ubc.ca
![]() |
Summary |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Tenascin-C (TN-C) and its isoforms are multidomain extracellular matrix (ECM) proteins that are believed to be involved in the regulation of stromalepithelial interactions. Some of the interactions between TN-C and cells are mediated by integrins. In this study we analyzed the expression of TN-C and its large molecular weight splice isoform (TN-CL) and the putative TN-C-binding 9 and
vß6 integrins during human wound repair. In 3-day-old oral mucosal wounds, immunoreactivity for
9 integrin localized abundantly at the migrating basal wound epithelial cells. TN-C and TN-CL were localized in the matrix between and underneath
9-expressing epithelial cells. In parallel with gradual downregulation of
9 integrin immunoreactivity in 7-day and older wounds, the expression of
vß6 integrin was temporarily induced. Integrin
vß6 co-localized in the same area as TN-C and TN-CL immunoreactivity at the cellcell contacts of the basal and suprabasal cell layers of the wound epithelium. During granulation tissue formation and reorganization from 7 to 28 days after wounding, TN-C and TN-CL were abundantly localized in the granulation tissue. The findings show that TN-CL is expressed under the migrating epithelial front and in the granulation tissue during matrix deposition in wound repair. Preferential localization of
9 integrin in migrating epithelial cells and of
vß6 integrin in epithelium after wound closure suggests different functions for these integrins in wound repair.
(J Histochem Cytochem 48:985998, 2000)
Key Words: integrins, tenascin-C, wound repair
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
WOUND HEALING is a complex process that in many cases resembles the events in development, and involves cell proliferation and migration along matrices that undergo remodeling. It is well documented that expression of cell adhesion receptors, integrins, and their ligands is important in the regulation of epithelial cell proliferation, differentiation, migration, and gene expression (
Tenascin (TN-C) is a large hexameric glycoprotein that is abundantly expressed, particularly at epithelialmesenchymal interaction sites during embryogenesis. TN-C shows a more restricted distribution in various adult tissues, including skin and oral mucosa, in which it localizes mostly to the papillary connective tissue immediately beneath the basement membrane zone (
Human TN-C is composed of structurally different domains, including epidermal growth factor-like repeats, units similar to fibronectin Type III homology repeat (TNfn), and a C-terminal sequence with homology to ß- and -chains of fibrinogen (
Cell adhesion to TN-C is mediated by integrins in a cell type-specific manner (- and ß-subunits. To date, 18
- and eight ß-subunits have been described, which can form at least 24 different
ß heterodimers (
vß6 and
9ß1 recognize the third TNfnIII (TNfn3). In TNfn3, however,
vß6 integrin recognizes the common integrin recognition motif Arg-Gly-Asp (RGD), whereas
9ß1 integrin binds to a sequence that includes the Ile-Asp-Gly (IDG) motif (
Previous studies have shown changes in the expression and distribution of epithelial integrins in wound repair compared with normal tissue (vß6 is not normally expressed in the epithelium but its expression is induced during wound repair (
9-subunit is a recently characterized member of the ß1 integrin family that is expressed by specialized cells, including stratified squamous epithelium in skin (
9-subunit is developmentally regulated and coincides with the development of stratification of epithelium (
In this study we tested our hypothesis that the expression of 9ß1 and
vß6 is coordinately upregulated with the large TN-C isoform during wound repair. We provide evidence that
9 but not ß6 integrin is expressed by migrating epithelial cells during early wound repair. This coincides with deposition of TN-C, including the large isoform, under the migrating epithelial cells. After epithelial confrontation, immunoreactivity for
9 integrin is reduced, whereas expression of
vß6 integrin is induced and it localizes in the same area as TN-C. During later tissue differentiation and maturation, immunoreactivity for
vß6 integrin and TN-C at the epithelial cell interfaces is coordinately reduced, whereas the immunoreactivity for TN-C is still strong in granulation tissue compared with normal connective tissue.
![]() |
Materials and Methods |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Histological Specimens
Tissue biopsy specimens of normal keratinized masticatory mucosa (gingiva, n = 10; alveolar mucosa, n = 3; palatal mucosa, n = 3) and non-keratinized buccal mucosa (n = 3) and ventral tongue (n = 1), were obtained from four healthy volunteers. Three-day, 7-, 14-, and 28-day-old wounds from keratinized palatal gingiva were obtained from the collection of wound sections from three volunteers previously used to localize integrins and basement membrane components during mucosal wound healing (
Antibodies
To localize different alternatively spliced TN-C isoforms, two monoclonal antibodies (MAbs) recognizing different TN-C epitopes were used (gifts from Dr. Luciano Zardi, Instituto Nazionale per la Ricerca sul Cancro, Genoa, Italy). MAb BC-4 recognizes an epitope within the EGF-like sequences that is common to all forms of TN-C. MAb BC-2 reacts with epitopes in the alternatively spliced FN-like repeats A1 and A4. In immunoblotting, MAb BC-2 recognizes the large TN-C isoform (280 kD) and MAb BC-4 the small and the large isoforms (190 kD and 280 kD, respectively). The characterization of the antibodies has been described previously (9 integrin subunit (
vß6 integrin (E7P6;
Immunohistochemical Staining
Immediately after biopsy, fresh tissue blocks were mounted in Histoprep (Fisher Scientific; Fair Lawn, NJ) and snap-frozen in liquid nitrogen. Frozen sections (6 µm) were cut and fixed with -20C acetone for 5 min and stored at -70C until used. For immunoperoxidase staining for integrin 9-subunit, sections were incubated in 0.3% hydrogen peroxide in methanol for 30 min and washed in PBS/BSA. Sections were rinsed and incubated with normal blocking serum (Vectastain; Vector Laboratories, Burlingame, CA) for 60 min at room temperature (RT) and then incubated with the primary antibody in PBS containing 0.1% bovine serum albumin (BSA; Sigma Chemical, St Louis, MO) in a humid chamber at 4C for 16 hr. After washing with PBS/BSA, sections were incubated with biotinylated anti-rabbit antibody for 60 min and then reacted with ABC avidinperoxidase reagent (Vectastain Elite kit; Vector Laboratories). Chromogen was developed using diaminobenzidine (DAB; Chemicon, Temecula, CA) and nickelsilver enhancement (
For immunofluorescence staining of vß6 integrin, sections were incubated for 60 min at RT with PBS/BSA and then with the primary antibody as above. After washing, sections were incubated with anti-mouse Igrhodamine (Boehringer Mannheim) in PBS/BSA for 60 min at RT. Sections were washed, briefly air-dried, and mounted with cyanoacrylate glue. Control sections were incubated with purified nonimmune rabbit or mouse IgG (10 µg/ml; Sigma) in PBS/BSA instead of the primary antibody (not shown).
Immunohistochemical staining for TN-C isoforms was performed using the alkaline phosphatasemonoclonal anti-alkaline phosphatase (APAAP) method. The primary antibodies BC-2 and BC-4 (undiluted hybridoma supernatant) were incubated for 30 min at RT. After washing with Tris buffer, sections were treated with rabbit anti-mouse immunoglobulin (diluted 1:70; Dako, Glostrup, Denmark) and then with the mouse APAAP complex (Dako). Both incubations were carried out for 30 min at RT. To increase the staining intensity, the incubation with the rabbit anti-mouse immunoglobulin and with the APAAP complex was repeated twice. Naphthol-AS-biphosphate (Sigma) and new fuchsin (Merck; Darmstadt, Germany) were used as substrate and developer, respectively. To inhibit endogenous tissue enzyme activity, the developing solution was supplemented with 0.25 mmol/liter levamisole (Sigma). After color development, the sections were counterstained briefly with hematoxylin and mounted in glycerol gelatin (Serva; Heidelburg, Germany). As negative control, the primary antibody was replaced by nonimmune serum (not shown).
Up to 10 sections of each sample were stained with each antibody. The staining was examined with a Zeiss Axioskop 20 light and fluorescence microscope and was photographed with an MC 80 Zeiss microscope camera. To evaluate the overall histology, including collagen fiber organization and degree of inflammation in the wound sections, a set of samples was stained with Mallory's phosphotungstic acidhematoxylin method (
![]() |
Results |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Localization of 9 and
vß6 Integrin and TN-C Splice Isoforms in Normal Keratinized and Non-keratinized Oral Mucosa
The expression of 9 integrin in keratinized epithelium of gingiva and alveolar and palatal mucosa was relatively weak. At epithelial rete ridges,
9 integrin was expressed by the basal epithelial cells and the immediately suprabasal cells, and was localized most abundantly against the basement membrane (Fig 1A). In the connective tissue,
9 integrin localized at blood vessels (Fig 1A). Immmunoreactivity for antibody BC-2, recognizing the large TN-C splice isoform, and for BC-4, recognizing all forms of TN-C, localized similarly at the epithelial basement membrane zone and over collagen fibers throughout the subepithelial connective tissue (Fig 1D and Fig 1E). The strongest staining of TN-C was localized at the ECM of the connective tissue papilla area. In the deep connective tissue matrix, the staining intensity for TN-C decreased (Table 1). Some of the subepithelial vascular basement membrane areas were stained with anti-TN-C antibodies (Fig 1D and Fig 1E).
|
|
In non-keratinized buccal mucosa (Fig 1B) and ventral tongue (Fig 1C), immunoreactivity for 9 integrin subunit was stronger compared with normal keratinized gingiva and showed most abundant staining at the cell membrane of the basal cells. In addition,
9 integrin had a pericellular distribution at the basal and suprabasal cell layers. In the connective tissue,
9 integrin localized at blood vessels (Fig 1B and Fig 1C). Strongest immunoreactivity for both BC-2 (Fig 1E and Fig 1F) and BC-4 (Fig 1H and Fig 1I) antibodies was localized at the epithelial and most of the vascular basement membranes. Staining intensity of the blood vessels with the BC-4 antibody was slightly more intense than with the BC-2 antibody. At connective tissue papilla areas and directly under the basement membrane, TN-C was localized on collagen fibers but, unlike in keratinized mucosa, no TN-C immunoreactivity was extended into the deeper areas of the subepithelial connective tissue (Fig 1E, Fig 1F, Fig 1H, and Fig 1I; Table 1). No expression of
vß6 integrin was seen in keratinized and nonkeratinized normal oral mucosa (not shown).
Localization of 9 and
vß6 Integrins and of TN-C Splice Isoforms During Wound Healing of Keratinized Oral Mucosa
In 3-day-old wounds, epithelium had started to migrate through the wound bed provisional matrix. A moderate inflammatory cell infiltrate was localized in the wound bed and in the connective tissue under the epithelium next to the wound. In 7-day-old wounds, epithelium had completely covered the wound space and had started to differentiate. The inflammation in the granulation tissue was reduced compared with earlier time points, and collagen deposition in the granulation tissue had started. At 14 days after wounding, only scattered inflammatory cells were seen in the granulation tissue. New collagen fibers in the granulation tissue were relatively thin compared with normal tissue, and were mostly arranged horizontally across the wound area. By Day 28, the inflammation had completely disappeared, but the granulation tissue could still be distinguished from the more organized normal connective tissue and was composed of thick collagen fibers that were mostly organized horizontally across the granulation tissue. At this time, epithelium had completely regenerated and was morphologically similar to adjacent unwounded epithelium.
In the epithelium immediately next to the wound margin, and in keratinocytes migrating through the provisional wound matrix of the 3-day-old wounds, immunoreactivity for 9 integrin was strong in the basal cells (Fig 2A and Fig 2B; Table 1). The most intense staining was localized at the basal cell membranes of the keratinocytes at the leading edge of the migrating epithelial front (Fig 2A and Fig 2B, large black arrowheads). In the wound fibrin clot,
9 immunoreactivity localized to scattered inflammatory cells (Fig 2A and Fig 2B, small arrows). In the unwounded connective tissue, immunoreactivity for
9 integrin localized abundantly at blood vessels (Fig 2A, open arrowheads). In 7-day-old wounds, in tissue sections from the central wound where migrating epithelial fronts had just joined to cover the wound surface, expression of
9 integrin at the basal membranes of the basal cells was still strong (Fig 2C and Fig 2D, large black arrowheads). However, in serial sections further away from the central wound where epithelium had already started differentiation, immunoreactivity for
9 integrin was weaker (Fig 2E and Fig 2F). Some forming blood vessels in the granulation tissue also showed positive staining for
9 integrin (Fig 2C and Fig 2E, open arrowheads). As maturation of the epithelium continued for 14 days after wounding, epithelium did not show any immunoreactivity for
9 integrin (Table 1). In contrast, blood vessels in the granulation tissue showed strong staining for
9 integrin (Table 1). After 28 days, expression of
9 integrin was normalized in the epithelium. Most abundant staining localized against the basement membrane, similar to normal unwounded tissue (Fig 2G and Fig 2H, large black arrowheads). Blood vessels at the subepithelial granulation tissue also showed positive staining for
9 integrin (Fig 2G and Fig 2H, open arrowheads).
|
No immunoreactivity for vß6 integrin was detected in the 3-day-old wounds (Fig 3A; Table 1). After 7 days, expression of
vß6 integrin was induced at the cell membranes of the basal and several suprabasal cell layers of the wound epithelium (Fig 3B). At Day 14 after wounding, expression of
vß6 integrin at the basal and immediately suprabasal cell layers was still strong (Fig 3C, inset), although its expression in the most suprabasal cell layers was reduced compared with the 7-day-old wounds (Fig 3C). After 28 days, the epithelium was totally negative for immunoreactivity for
vß6 integrin (Fig 3D; Table 1).
|
After 3 days, immunoreactivity for both BC-2 (Fig 4A and Fig 4B) and BC-4 (Fig 5A and Fig 5B) anti-TN-C antibodies was similar and was localized around the cell membranes of the migrating basal epithelial cells, but no TN-C staining was seen in the provisional matrix of the wound bed (Fig 4A, Fig 4B, Fig 5A, and Fig 5B; Table 1). At the cellcell interfaces of the basal and some suprabasal cell layers in the epithelium next to the wound, immunoreactivity for both BC-2 (Fig 4A and Fig 4B) and BC-4 (Fig 5A and Fig 5B) antibodies also localized between the basal and immediately suprabasal epithelial cells. In the nonwounded area further away from the wound, TN-C immunoreactivity between the epithelial cells was gradually suppressed (Fig 4A and Fig 5A). After 7 days, immunoreactivity for BC-2 (Fig 4C and Fig 4D) and BC-4 (Fig 5C and Fig 5D) antibodies was similar and was localized at the cellcell interfaces at the basal and suprabasal cell layers of the wound epithelium. Furthermore, immunoreactivity for both BC-2 (Fig 4C) and BC-4 (Fig 5C) antibodies was relatively strong throughout the wound granulation tissue compared with adjacent normal connective tissue (Table 1). After 14 days of wounding, immunoreactivity for both BC-2 (Fig 4E and Fig 4F) and BC-4 (Fig 5E and Fig 5F) antibodies was similarly reduced between the suprabasal epithelial cell layers and was localized only around cell membranes of the basal cells. The granulation tissue showed similar strong immunoreactivity for both BC-2 (Fig 4E and Fig 4F) and BC-4 (Fig 5E and Fig 5F) antibodies. After 28 days, immunoreactivity for both BC-2 (Fig 4G and Fig 4H) and BC-4 (Fig 5G and Fig 5H) antibodies was no longer localized at the cellcell interfaces at the wound epithelium, although their staining intensity was still relatively strong throughout the wound granulation tissue (Table 1).
|
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
By using different human and animal wound healing models, previous studies have documented increased TN-C deposition and expression during wound repair (
Cell culture and in situ hybridization findings showing that migratory keratinocytes express TN-C mRNA in early wound repair suggest that, at this point, TN-C is produced by the migratory keratinocytes (-smooth muscle actin is induced in fibroblasts that are actively involved in cell-mediated wound contraction (
-smooth muscle actin in fibroblasts at 7-day-old wounds, with a peak expression at 14 days after wounding (unpublished observation), which coincided with the strong TN-C expression in the granulation tissue.
During wound repair, immunoreactivity for the integrin 9-subunit was upregulated at the basal cell membrane of the migrating basal epithelial cells in 3- and 7-day-old wounds. Interestingly, in areas of 7-day-old wounds at which epithelial migration had stopped and epithelium had started to differentiate, and in 14-day-old wounds in which epithelial maturation was under way, expression of
9 integrin was relatively weak by the epithelial cells. This suggests that
9 integrin functions in cell migration in early wounds, whereas its expression must be downregulated during epithelial differentiation. In corneal wound repair,
9 integrin also localizes to the migrating corneal epithelial cells (
9 integrin expression coincides with epithelial stratification (
9 integrin, suggesting that
9 integrin has a similar function during embryonic development and wound repair.
The expression of 9 integrin in migrating epithelial cells coincides with expression of TN-C under the migrating epithelial front and co-localizes with integrin ß1-subunit during early wound repair (
9ß1 integrin may be one of the cell surface receptors that mediates cell migration on TN-C-containing provisional matrix during wound re-epithelialization. On the other hand, in addition to regulating cell migration, TN-C can also induce cell proliferation. The mitogenic activity appears to be associated with a region in the fibronectin Type III domain that is recognized by
9ß1 integrin (
9ß1 integrin in cells plated on this TN-C fragment induces cell proliferation (
9 integrin and TN-C may be one of the mechanisms that regulate epithelial cell proliferation during wound repair.
Downregulation of 9 integrin in the wound epithelium coincided with induced expression of
vß6 integrin from Day 7 to Day 14 after wounding. The expression of
vß6 integrin temporarily and spatially overlapped with the expression of TN-C at the cellcell interfaces at the wound epithelium. During granulation tissue reorganization and remodeling between Days 7 and 28, the expression of
vß6 integrin and TN-C in the wound epithelium was gradually downregulated. Expression of integrins at the cellcell contacts is a common finding, although no ligands for integrins are normally present. Co-localization of TN-C and
vß6 integrin at the cellcell interfaces at the wound epithelium suggest that, at least during wound repair, integrins may also bind their ligands at the epithelial cell interfaces. The finding that heterologous expression of
9ß1 integrin in cells plated on TN-C induces, whereas ß6 integrin downregulates, cell proliferation (
9 and
vß6 integrins in the wound epithelium may be one of the mechanisms that regulate cell responses during epithelial differentiation.
Previous studies after wound repair up to Day 7 after wounding have demonstrated induced vß6 integrin expression by wound epithelial cells (
vß6 integrin was detected in 7-day-old wounds. However, after 14 days, expression of
vß6 integrin was still upregulated in the epithelium covering the granulation tissue. At 28 days after wounding, when the organization of the wound granulation tissue was more complete, the wound epithelium became completely negative for
vß6 immunoreativity. Findings from studies using mice with targeted disruption or constitutive overexpression of ß6 integrin gene indicate that epithelial
vß6 integrin expression may downregulate inflammation in the underlying connective tissue (
vß6 integrin to activate latent TGF-ß (
vß6 integrin in the epithelium.
Our findings in this study showed expression of 9 integrin by inflammatory cells in the provisional matrix of the blood clot after 3 days of wounding. Recent findings have shown that
9ß1 integrin selectively expressed on neutrophils mediates their adhesion on activated endothelial cells through binding to VCAM-1, which mediates chemotaxis of neutrophils on endothelial cells (
9 integrin in the extravasation of neutrophils in the acute inflammatory phase of wound repair.
The integrin 9-subunit was relatively highly expressed in basal and suprabasal cells of normal non-keratinized epithelium of ventral tongue and buccal mucosa, whereas it showed a weaker expression in keratinized epithelium of gingiva, in which it localized mostly to the basal cells. The regional variations in keratinization and differentiation of oral mucosa affect also the expression pattern of other integrins in the oral epithelium. In normal keratinized oral mucosa, expression of integrins is mainly confined to the basal epithelial cells that express
2ß1,
3ß1,
6ß4, and
vß5 integrins (
9 integrin in human nonkeratinized mucosa may be associated with enhanced proliferation of the epithelial cells. In fact, in conditions such as wound healing or psoriasis, rapid cell proliferation is associated with increased integrin expression in suprabasal cell populations (
9 in these processes is identical to that of ß1 integrin (
9ß1 integrin may play a role in the regulation of cell proliferation and differentiation during tissue maintenance.
In the basal epithelial cells of normal keratinized and nonkeratinized oral mucosa, 9 integrin showed the most abundant staining against the basement membrane. TN-C immunoreactivity for both BC-2 and BC-4 antibodies was also localized at the basement membrane zone in all areas of oral mucosa, indicating that the large TN-C isoform is also normally expressed in oral mucosa. Electron microscopic localization of TN-C in rat skin indicates that TN-C is actually a component of the basement membrane (
9 integrin. Interestingly, TN-C showed differential localization in the connective tissue of keratinized gingiva and in nonkeratinized buccal and ventral tongue mucosa. In gingiva, TN-C was expressed throughout the connective tissue matrix but in only few vascular basement membranes. In contrast, TN-C was mainly localized at the epithelial and vascular basement membranes in the nonkeratinized oral mucosa but relatively weak staining was detected in the connective tissue. It is possible that the differential TN-C expression in various regions of oral mucosa may reflect functional differences in these tissues.
Previous findings have shown that 9 integrin is temporarily expressed in vascular smooth muscle cells during murine development (
9 integrin is not usually expressed by endothelial cells during development or in adult tissue (
9 integrin at the blood vessels of normal oral mucosal connective tissue and in wound granulation tissue. Although we did not specifically show if
9 integrin expression localized to vascular smooth muscle cells or endothelial cells, the finding that
9 integrin is expressed at blood vessels in adult human oral mucosa and in wound granulation tissue is new, and it suggests that vascular expression of
9 integrin may be regulated in a tissue-specific manner.
Taken together, our findings suggest that interaction between 9 integrin and TN-C isoforms, including the large molecular weight isoform, may play a role in the maintenance and differentiation of normal keratinized and nonkeratinized oral mucosa. Furthermore, the finding that TN-C isoforms and their putative receptors,
9 and
vß6 integrins, are spatially and temporally coordinately expressed during wound repair, suggest that interactions of these integrins with TN-C may be among mechanisms that regulate epithelial cell proliferation, migration, and phenotype during wound repair.
![]() |
Acknowledgments |
---|
Supported by grants from the Medical Research Council of Canada.
We would like to thank Cristian Sperantia, Andre Wong and Bruce McCaughey for expert technical assistance.
Received for publication November 2, 1999; accepted February 16, 2000.
![]() |
Literature Cited |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Aukhil I, Sahlberg C, Thesleff I (1996) Basal layer of epithelium expresses tenascin mRNA during healing of incisional skin wounds. J Periodont Res 31:105-112[Medline]
Balza E, Siri A, Ponassi M, Caocci F, Linnala A, Virtanen I, Zardi L (1993) Production and characterization of monoclonal antibodies specific for different epitopes of human tenascin. FEBS Lett 332:39-43[Medline]
Borsi L, Balza E, Castellani P, Carnemolla B, Ponassi M, Querze G, Zardi L (1994) Cell-cycle dependent alternative splicing of the tenascin primary transcript. Cell Adhes Commun 1:307-317[Medline]
Borsi L, Carnemolla B, Nicolo C, Spina B, Tanara G, Zardi L (1992) Expression of different tenascin isoforms in normal, hyperplastic and neoplastic breast tissues. Int J Cancer 52:688-692[Medline]
Brakebusch C, Hirsch E, Potocnik A, Fassler R (1997) Genetic analysis of beta 1 integrin function: confirmed, new and revised roles for crucial family of cell adhesion molecules. J Cell Sci 110:2895-2904
Breuss JM, Gallo J, DeLisser HM, Klimanskaya IV, Folkesson HG, Pittet JF, Nishimura SL, Aldape K, Landers DV, Carpenter W, Gillett N, Sheppard D, Matthay MA, Abelda SM, Kramer RH, Pytela R (1995) Expression of the ß6 integrin subunit in development, neoplasia and tissue repair suggests a role in epithelial remodelling. J Cell Sci 108:2241-2251
Camper L, Hellman U, Lundgren-Akerlund E (1998) Isolation, cloning, and sequence analysis of the integrin subunit 10, a ß1-associated collagen binding integrin expressed on chondrocytes. J Biol Chem 273:20383-20389
ChiquetEhrismann R, Hagios C, Schenk S (1995) The complexity in regulating the expression of tenascin. BioEssays 17:873-878[Medline]
ChiquetEhrismann R, Mackie EJ, Pearson CA, Sakakura T (1986) Tenascin: an extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis. Cell 47:131-139[Medline]
ChiquetEhrismann R, Matsuoka Y, Hofer U, Spring J, Bernasconi C, Chiquet M (1991) Tenascin variants: differential binding to fibronectin and distinct distribution in cell cultures and tissues. Cell Regul 2:927-938[Medline]
ChiquetEhrismann R, Tannheimer M, Koch M, Brunner A, Spring J, Martin D, Baumgartner S, Chiquet M (1994) Tenascin-C expression by fibroblasts is elevated in stressed collagen gels. J Cell Biol 127:2093-2101[Abstract]
Chuong C-M, Chen H-M (1991) Enhanced expression of neural cell adhesion molecules and tenascin (cytotactin) during wound healing. Am J Pathol 138:427-440[Abstract]
Clark RAF (1993) Regulation of fibroplasia in cutaneous wound repair. Am J Med Sci 306:42-48[Medline]
Crossin KL (1996) Tenascin: a multifunctional extracellular matrix protein with a restricted distribution in development and disease. J Cell Biochem 61:592-598[Medline]
Darby I, Skalli O, Gabbiani G (1990) -Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21-29[Medline]
Desloges N, Basora N, Perrault N, Bouatrouss Y, Sheppard D, Beaulieu J-F (1998) Regulated expression of the integrin 9ß1 in the epithelium of the developing human gut and in intestinal cell lines: relation with cell proliferation. J Cell Biochem 71:536-545[Medline]
End P, Panayotou G, Entwistle A, Waterfield MD, Chiquet M (1992) Tenascin: a modulator of cell growth. Eur J Biochem 209:1041-1051[Abstract]
Fisher D, Brown-Ludi M, Schulthess T, ChiquetEhrismann R (1997a) Concerted action of tenascin-C domains in cell adhesion, anti-adhesion and promotion of neurite growth. J Cell Sci 110:1513-1522
Fisher D, Tucker RP, ChiquetEhrismann R, Adams JC (1997b) Cell-adhesive responses to tenascin-C splice variants involve formation of fascin microspikes. Mol Biol Cell 8:2055-2057
Haapasalmi K, Zhang K, Tonnesen M, Olerud J, Sheppard D, Salo T, Kramer R, Clark RA, Uitto VJ, Larjava H (1996) Keratinocytes in human wounds express vß6 integrin. J Invest Dermatol 106:42-48[Abstract]
Hertle MD, Kubler MD, Leigh IM, Watt FM (1992) Aberrant integrin expression during epidermal wound healing and in psoriatic epidermis. J Clin Invest 89:1892-1901[Medline]
Huang XZ, Wu JF, Cass D, Erle DJ, Corry D, Young SG, Farese RV, Jr, Sheppard D (1996) Inactivation of the integrin ß6 gene reveals a role of epithelial integrins in regulating inflammation in the lungs and skin. J Cell Biol 133:921-928[Abstract]
Huang X, Wu J, Zhu W, Pytela R, Sheppard D (1998) Expression of the human integrin ß6 subunit in alveolar type II cells and bronchiolar epithelial cells reverses lung inflammation in ß6 knock out mice. Am J Respir Cell Mol Biol 19:636-642
Jones J, Sugiyama M, Watt FM, Speight PM (1993) Integrin expression in normal, hyperplastic, dysplastic, and malignant oral epithelium. J Pathol 169:235-243[Medline]
Jones J, Watt FM, Speight PM (1997) Changes in the expression of alpha v integrins in oral squamous cell carcinomas. Oral Pathol Med 26:63-68[Medline]
Jones PH, Harper S, Watt FM (1995) Stem cell patterning and fate in human epidermis. Cell 80:83-93[Medline]
Juhasz I, Murphy GF, Yan HC, Herlyn M, Albelda SM (1993) Regulation of extracellular matrix proteins and integrin cell substratum adhesion receptors on epithelium during cutaneous human wound healing in vivo. Am J Pathol 143:1458-1469[Abstract]
Kaplony A, Zimmermann DR, Fisher RW, Imhof BA, Odermatt BF, Winterhalter KH, Vaughan L (1991) Tenascin Mr 220,000 isoform expression correlates with corneal cell migration. Development 112:605-614[Abstract]
Katz BZ, Yamada KM (1997) Integrins in morphogenesis and signaling. Biochimie 79:467-476[Medline]
Kruse J, Keihauer G, Faissner A, Timpl R, Scharchner M (1985) The J1 glycoprotein: a novel nervous system cell adhesion molecule of the L2/HNK-1 family. Nature 316:146-148[Medline]
Larjava H, Salo T, Haapasalmi K, Kramer RH, Heino J (1993) Expression of integrins and basement membrane components by wound keratinocytes. J Clin Invest 92:1425-1435[Medline]
Latijnhouwers MA, Bergers M, Ponec M, Dijkman H, Andriessen M, Schalkwijk J (1997) Human epidermal keratinocytes are a source of tenascin-C during wound healing. J Invest Dermatol 108:776-783[Abstract]
Latijnhouwers MA, Bergers M, Van Bergen BH, Spruijt KI, Andriessen MP, Schalkwiljk J (1996) Tenascin expression during wound healing in human skin. J Pathol 178:30-35[Medline]
Lukinmaa P-L, Allemanni G, Waltimo J, Zardi L (1996) Immunoreactivity of tenascin-C in dentin matrix in dentinogenesis imperfecta associated with osteogenesis imperfecta. J Dent Res 75:581-587[Abstract]
Luomanen M, Virtanen I (1993) Distribution of tenascin in healing incision, excision and laser wounds. J Oral Pathol Med 22:41-45[Medline]
Mackie EJ, Halfter W, Liverani D (1988) Induction of tenascin in healing wounds. J Cell Biol 107:2757-2767[Abstract]
Mackie EJ, Thesleff I, ChiquetEhrismann R (1987) Tenascin is associated with chondrogenic and osteogenic differentiation in vivo and promotes chondrogenesis in vitro. J Cell Biol 105:2569-2579[Abstract]
Matsuoka Y, Spring J, BallmerHofer K, ChiquetEhrismann R (1990) Differential expression of tenascin splicing variants in the chick gizzard and in cell cultures. Cell Differ Dev 32:417-424[Medline]
Mighell AJ, Thompson J, Hume WJ, Markham AF, Robinson PA (1997) Human tenascin-C: identification of a novel type III repeat in oral cancer and of novel splice variants in normal, malignant and reactive oral mucosae. Int J Cancer 72:236-240[Medline]
Moles J-P, Watt FM (1997) The epidermal stem cell compartment: variation in expression levels of E-cadherin and catenins within the basal layer of human epidermis. J Histochem Cytochem 45:867-874
Munger JS, Huang X, Kawakatsu H, Griffiths MJ, Dalton SL, Wu J, Pittet JF, Kaminski N, Garat C, Matthay MA, Rifkin DB, Sheppard D (1999) The integrin alpha v beta 6 binds and activates latent TGFß1: a mechanism for regulating pulmonary inflammation and fibrosis. Cell 96:319-328[Medline]
MurphyUllrich JE, Lightner VA, Aukhil I, Yan YZ, Erickson HP, Hook M (1991) Focal adhesion integrity is downregulated by the alternatively spliced domain of human tenascin. J Cell Biol 115:1127-1136[Abstract]
Palmer EL, Ruegg C, Ferrando R, Pytela R, Sheppard D (1993) Sequence and tissue distribution of the integrin 9 subunit, a novel partner of ß1 that is widely distributed in epithelia and muscle. J Cell Biol 123:1289-1297[Abstract]
Phillips GR, Krushel LA, Crossin KL (1998) Domains of tenascin involved in glioma migration. J Cell Sci 111:1095-1104
Prieto AL, AnderssonFisone C, Crossin KL (1992) Characterization of multiple adhesive and counteradhesive domains in the extracellular matrix protein cytotactin. J Cell Biol 119:663-678[Abstract]
Prieto AL, Jones FS, Cunningham BA, Crossin KL, Edelman GM (1990) Localization during development of alternatively spliced forms of cytotactin mRNA by in situ hybridization. J Cell Biol 111:685-698[Abstract]
Przepiorka D, Myerson D (1986) A single-step silver enhancement method permitting rapid diagnosis of cytomegalovirus infection in formalin-fixed, paraffin-embedded tissue sections by in situ hybridization and immunoperoxidase detection. J Histochem Cytochem 34:1731-1734[Abstract]
Puchtler H, Sweat F, Doss NO (1963) A one-hour phosphotungstic acid-hematoxylin stain. Am J Clin Pathol 40:334-337[Medline]
Ramos DM, Chen BL, Boylen K, Stern M, Kramer RH, Sheppard D, Nishimura SL, Greenspan D, Zardi L, Pytela R (1997) Stromal fibroblasts influence oral squamous-cell carcinoma cell interactions with tenascin-C. Int J Cancer 74:369-376
Rettig WJ, Erickson HP, Albino AP, GarinChesa P (1994) Induction of human tenascin (neuronectin) by growth factors and cytokines: cell type-specific signals and signaling pathways. J Cell Sci 107:487-497
Rikimaru K, Moles J-P, Watt FM (1997) Correlation between hyperproliferation and suprabasal integrin expression in human epidermis reconstituted in culture. Exp Dermatol 6:214-221[Medline]
Rowat JS, Squier CA (1986) Rates of epithelial cell proliferation in the oral mucosa and skin of the Tamarin monkey (Saguinus fuscicollis). J Dent Res 65:1326-1331[Abstract]
Schenk S, BrucknerTuderman L, Chiquet-Ehrismann R (1995) Dermo-epidermal separation is associated with induced tenascin expression in human skin. Br J Dermatol 133:13-22[Medline]
Siri A, Carnemolla B, Saginati M, Leprini A, Casari G, Barella F, Zardi L (1991) Human tenascin primary structure, pre-mRNA splicing patterns and localization of the epitopes recognized by two monoclonal antibodies. Nucleic Acids Res 19:525-531[Abstract]
Stepp MA, Zhu L (1997) Upregulation of 9 integrin and tenascin during epithelial regeneration after debridement in the cornea. J Histochem Cytochem 45:189-201
Stepp MA, Zhu L, Sheppard D, Cranfill RL (1995) Localized distribution of 9 integrin in the cornea and changes in expression during corneal epithelial cell differentiation. J Histochem Cytochem 43:353-362
Taooka Y, Chen J, Yednock T, Sheppard D (1999) The integrin 9ß1 mediates adhesion to activated endothelial cells and transendothelial neutrophil migration through interaction with vascular cell adhesion molecule-1. J Cell Biol 145:413-420
Thorup AK, Dabelsteen E, Schou S, Gil SG, Carter WG, Reibel J (1997) Differential expression of integrins and laminin-5 in normal oral epithelia. APMIS 105:519-530[Medline]
Tucker RP, Hammarback JA, Jenrath DA, Mackie EJ, Xu Y (1993) Tenascin expression in the mouse: in situ localization and induction in vitro by bFGF. J Cell Sci 104:69-76
Wang A, Patrone L, McDonald JA, Sheppard D (1995) Expression of the integrin subunit 9 in the murine embryo. Dev Dyn 204:421-431[Medline]
Weinacker A, Chen A, Agrez M, Cone RI, Nishimura S, Wayner E, Pytela R, Sheppard D (1994) Role of integrin vß6 in cell attachment to fibronectin: heterologous expression of intact and secreted forms of the receptor. J Biol Chem 269:6940-6948
Whitby DJ, Longaker MT, Harrison MR, Adzick NS, Ferguson MWJ (1991) Rapid epithelialization of fetal wounds is associated with the early deposition of tenascin. J Cell Sci 99:583-586[Abstract]
Yokosaki Y, Matsuura N, Higashiyama S, Murakami I, Obara M, Yamakido M, Shigeto N, Chen J, Sheppard D (1998) Identification of the ligand binding site for the integrin 9ß1 in the third fibronectin type III repeat of tenascin-C. J Biol Chem 273:11423-11428
Yokosaki Y, Monis H, Chen J, Sheppard J (1996) Differential effects of the integrins 9ß1,
vß3, and
vß6 on cell proliferative responses to tenascin. J Biol Chem 271:24144-24150
Yokosaki Y, Palmer EL, Prieto AL, Crossin KL, Bourdon MA, Pytela R, Sheppard D (1994) The integrin 9ß1 mediates cell attachment to a non-RGD site in the third fibronectin type III repeat of tenascin. J Biol Chem 269:26691-26696
Zhao Y, Young SL (1995) TGF-beta regulates expression of tenascin alternative-splicing isoforms in fetal lung. Am J Physiol 268:L173-180