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I Thesleff

Publications and source records attributed to I Thesleff.

At least 37 records · Page 2Linked to original sources
PubMed1991

Advances in histological methods open up new perspectives for craniofacial growth research.

Changes in the tissue architecture and composition which accompany growth and development have traditionally been mapped using histological methods. The modern technology now offers new possibilities which can be used in connection with light microscopy and electron microscopy. These techniques, particularly immunohistochemistry and in situ hybridization allow studies on spatial and temporal changes in molecular patterns of substances during tissue organization and cell differentiation. In this paper, we describe the principles of immunohistology, autoradiography, and in situ hybridization, and review our recent studies in which we have applied such methods to assess craniofacial development. We have used autoradiography to localize cell proliferation and epidermal growth factor (EGF) receptors in the developing mandibular condyle. We have also used immunohistochemistry to localize the extracellular matrix component, tenascin, in differentiating chondrocytes of the mandibular condyle, employing specific antibodies. In studies on the odontogenic potential of dental papilla mesenchyme, we have used hybridoma technology, and produced monoclonal antibodies against dental papilla mesenchyme. Most recently, we have used in situ hybridization, which allows detection of gene expression in tissue sections to localize transcripts of a fibroblast-growth-factor related gene (Int-2 proto-oncogene) in developing teeth. In future, successful combination of such new histological methods and traditional experimental procedures can be expected to produce new answers to the questions about the regulation of craniofacial growth.

Facial Bones↗
PubMed1990

Comparison of the distribution patterns of tenascin and alkaline phosphatase in developing teeth, cartilage, and bone of rats and mice.

Tenascin is a glycoprotein of the extracellular matrix, which has been associated with differentiation of hard tissue forming cells. Alkaline phosphatase (AP) is involved in calcification, and it has also been suggested to function in cell differentiation. We have compared the distributions of tenascin and AP in the developing skull and teeth of embryonic and growing rats and mice. Tenascin was localized by immuno-Peroxidase and AP by enzyme histochemical staining of tissue sections. Both tenascin and AP were largely restricted to bone, cartilage, and teeth. In cartilage, tenascin was expressed in the perichondrium, whereas AP activity was detected only in the hypertrophic cartilage. In growing intramembranous bone, tenascin and AP were expressed in the periosteum and endosteum. AP activity was restricted to the inner layer of the periosteum, whereas tenascin expression extended to the more superficial layers. In bud-staged teeth tenascin but no AP activity was localized in the condensing mesenchymal cells around the epithelial bud. At the bell stage both tenascin and AP activity were localized in the cuspal mesenchyme, and the intensity of staining decreased towards the cervical region. In summary, tenascin was present at all sites of AP activity except in the epithelial cells of the enamel organ and the hypertrophic cartilage of the mandibular condyle. In mesenchymal tissues tenascin was more widely distributed than AP. It can be suggested that tenascin has functions at earlier stages of hard tissue formation than AP.

Alkaline Phosphatase↗
PubMed1990

Molecular changes during determination and differentiation of the dental mesenchymal cell lineage.

The lineage of dental mesenchymal cells originates in the cranial neural crest, and after sequential determination and differentiation, gives rise to all structures of the tooth and its supporting tissues, except the enamel. Reciprocal interactions between the epithelial and mesenchymal tissues are conceivably the most important regulators of dental mesenchymal cell differentiation. The molecular mechanisms of this epigenetic regulation are not known at present. In order to examine the mechanisms of regulation of gene expression in the lineage of dental mesenchymal cells, information is needed on the molecular changes that accompany advancing differentiation. By using the molar tooth germ of mouse embryos as a model system, the changes in the expression of some molecules have been analysed by immunohistological localization and in situ hybridization, and the roles of tissue interactions in this process examined. This has shown that syndecan, a recently characterized cell surface proteoglycan, and tenascin, a matrix glycoprotein, appear in the condensing dental mesenchyme during the bud stage of tooth development. During the cap stage, dental mesenchyme is characterized by continued intense expression of syndecan, but this is lost during terminal differentiation of odontoblasts. Tenascin and syndecan may mediate cell-matrix interactions during condensation of dental mesenchymal cells. Expression of the Int-2 proto-oncogene, coding for a fibroblast growth factor-related molecule, can be detected by in situ hybridization in dental mesenchyme at the cap stage. This expression persists in cuspal mesenchyme at the bell stage but is lost from odontoblasts and from pulpal mesenchyme at progressive stages of tooth development. The advancement of tooth morphogenesis from cap to bell stage is accompanied by expression of alkaline phosphatase in the cuspal mesenchyme. Also tenascin, which is only weakly expressed during the cap stage, appears in the cuspal areas and shows codistribution with alkaline phosphatase. These observations indicate that the sequential determination and differentiation of the dental mesenchymal cells are characterized by a cascade of specific molecular changes. The cell surface proteoglycan syndecan and the Int-2 proto-oncogene are specific and transient markers of early dental mesenchymal cell differentiation. This information allows studies on the mechanisms of developmental regulation. These experimental tissue recombination studies indicate that the expression of syndecan and tenascin in the early dental mesenchyme is induced by the presumptive dental epithelium.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Syndecan and tenascin expression is induced by epithelial-mesenchymal interactions in embryonic tooth mesenchyme.

Morphogenesis of embryonic organs is regulated by epithelial-mesenchymal interactions associating with changes in the extracellular matrix (ECM). The response of the cells to the changes in the ECM must involve integral cell surface molecules that recognize their matrix ligand and initiate transmission of signal intracellularly. We have studied the expression of the cell surface proteoglycan, syndecan, which is a matrix receptor for epithelial cells (Saunders, S., M. Jalkanen, S. O'Farrell, and M. Bernfield. J. Cell Biol. In press.), and the matrix glycoprotein, tenascin, which has been proposed to be involved in epithelial-mesenchymal interactions (Chiquet-Ehrismann, R., E. J. Mackie, C. A. Pearson, and T. Sakakura. 1986. Cell. 47:131-139) in experimental tissue recombinations of dental epithelium and mesenchyme. Our earlier studies have shown that in mouse embryos both syndecan and tenascin are intensely expressed in the condensing dental mesenchyme surrounding the epithelial bud (Thesleff, I., M. Jalkanen, S. Vainio, and M. Bernfield. 1988. Dev. Biol. 129:565-572; Thesleff, I., E. Mackie, S. Vainio, and R. Chiquet-Ehrismann. 1987. Development. 101:289-296). Analysis of rat-mouse tissue recombinants by a monoclonal antibody against the murine syndecan showed that the presumptive dental epithelium induces the expression of syndecan in the underlying mesenchyme. The expression of tenascin was induced in the dental mesenchyme in the same area as syndecan. The syndecan and tenascin positive areas increased with time of epithelial-mesenchymal contact. Other ECM molecules, laminin, type III collagen, and fibronectin, did not show a staining pattern similar to that of syndecan and tenascin. Oral epithelium from older embryos had lost its ability to induce syndecan expression but the presumptive dental epithelium induced syndecan expression even in oral mesenchyme of older embryos. Our results indicate that the expression of syndecan and tenascin in the tooth mesenchyme is regulated by epithelial-mesenchymal interactions. Because of their early appearance, syndecan and tenascin may be used to study the molecular regulation of this interaction. The similar distribution patterns of syndecan and tenascin in vivo and in vitro and their early appearance as a result of epithelial-mesenchymal interaction suggest that these molecules may be involved in the condensation and differentiation of dental mesenchymal cells.

Animals↗
PubMed1989

Growth factors and tooth development.

The effects of various growth factors on tooth development were studied in organ cultures of mouse embryonic tooth germs. Transferrin was shown to be a necessary growth factor for early tooth morphogenesis. Transferrin was required for the development of bud- and early cap-staged teeth, and it was shown to be the only serum protein that was needed by early cap-staged teeth in organ culture. Promotion of tooth morphogenesis and dental cell differentiation was shown to be based on the stimulation of cell proliferation. The roles of polypeptide growth factors in tooth development were studied by adding these factors to the transferrin-containing chemically-defined culture medium which supports early tooth morphogenesis and cell differentiation. Fibroblast growth factor or platelet-derived growth factor did not affect cell proliferation or morphogenesis of tooth germs in culture. On the contrary, epidermal growth factor (EGF) stimulated cell proliferation in tooth explants, but at the same time inhibited tooth morphogenesis and dental cell differentiation. Autoradiographic localization of proliferating cells revealed that dental tissues responded to EGF with different proliferation rates. The responsiveness to EGF was stage-dependent, early cap-staged teeth were sensitive to EGF but late cap-staged and bell-staged teeth developed normally in the presence of EGF in the culture medium. The presence and distribution of receptors for both transferrin and EGF were studied in mouse embryonic teeth at various developmental stages by incubating freshly-separated tooth germs with 125Iodine-labeled transferrin or EGF, and then processing the tissues for autoradiography.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Cell-matrix interactions in tooth development.

A chain of reciprocal interactions between the epithelial and mesenchymal tissues regulates both morphogenesis and cell differentiation in the developing tooth. The very early interactions lead to budding of the oral epithelium and to the characteristic condensation of the neural crest-derived mesenchymal cells around the epithelial bud. During the bell stage of morphogenesis, the mesenchymal cells which are in contact with the dental epithelium differentiate into odontoblasts. In this reveiw article we summarize the results of our descriptive and experimental studies, which indicate that differentiation of the dental mesenchymal cells into odontoblasts, as well the condensation of dental mesenchymal cells at the bud stage, are regulated by interactions between the cell surface and the extracellular matrix. Transfilter studies where the dental epithelium and mesenchyme were cultured on opposite sides of Nuclepore filters, led to the hypothesis that the differentiation of dental mesenchymal cells into odontoblasts is triggered by interactions between the cell surface and the epithelial basement membrane matrix. Immunohistochemical localization of various matrix molecules showed that the matrix glycoproteins fibronectin and tenascin are accumulated in the dental basement membrane at the time of odontoblast differentiation. Fibronectin and tenascin are known to interact with each other, with other matrix molecules as well as with the cell surface, and also to influence cell shape. We suggest that fibronectin and tenascin are involved in the cell-matrix interaction which leads to the polarization and differentiation of odontoblasts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Immunohistochemical localization of the matrix glycoprotein tenascin in the skull of the growing rat.

Tenascin is an extracellular matrix glycoprotein which interacts with other matrix molecules and with cells, and which appears to play important roles in growth and differentiation. The immunohistochemistry of sections of newborn, 5-day and 20-day-old rats showed that accumulation of tenascin was largely restricted to bones, cartilages and teeth. It was also present in the periosteal and endosteal surfaces of membrane bones, in perichondrium, and ion the dental pulp, but was absent from mature bone, cartilage and dentine. In nasal cartilage, tenascin was present only in the perichondrium, whereas in the condylar cartilage staining was observed in the proliferating and maturing cell layers but not in the hypertrophied cartilage. These differences may reflect differences in the growth mechanisms of primary and secondary cartilages. Accumulation of tenascin was particularly striking in areas where the periosteum or perichondrium was thickened such as sites of some muscle attachments, sutures and condylar cartilage. The restricted distribution of tenascin is unlike the patterns observed for other extracellular matrix molecules. Tenascin may have a unique role in bone growth and remodelling in the craniofacial region.

Animals↗
PubMed1988

Cell surface proteoglycan expression correlates with epithelial-mesenchymal interaction during tooth morphogenesis.

Tooth morphogenesis and differentiation of the dental cells are guided by interactions between epithelial and mesenchymal tissues. Because the extracellular matrix is involved in these interactions, the expression of matrix receptors located at the cell surface may change during this developmental sequence. We have examined the distribution of an epithelial cell surface proteoglycan antigen, known to behave as a receptor for interstitial matrix, during tooth morphogenesis. Intense staining was seen around the cells of the embryonic oral epithelium as well as the dental epithelium at the early bud stage. With development, expression was greatly reduced in the enamel organ. Differentiation of these cells into ameloblasts was associated with the loss of expression, while the epithelial cells remaining in the stratum intermedium and stellate reticulum regained intense staining. The PG antigen was weakly expressed in the loose neural crest-derived jaw mesenchyme but it became strongly reactive in the condensed dental papilla mesenchyme when extensive morphogenetic movements took place. With development, the PG antigen disappeared from the advanced dental papilla mesenchyme but persisted in the dental sac mesenchyme, which gives rise to periodontal tissues. The PG antigen was not expressed by odontoblasts. Hence, the expression of the PG antigen changes during the epithelial-mesenchymal interactions of tooth development and is lost during terminal cell differentiation. The expression follows morphogenetic rather than histologic boundaries. The acquisition and loss of expression in epithelial and mesenchymal tissues during tooth development suggest that this proteoglycan has specific functions in the epithelial-mesenchymal interactions that guide morphogenesis.

Animals↗
PubMed1988

The parotid gland is the main source of human salivary epidermal growth factor.

To clarify the production of human epidermal growth factor (EGF) by different salivary glands, we measured its concentration by radioimmunoassay separately in whole saliva, in parotid gland (PG) saliva and in mixed submandibular (SMG) and sublingual gland (SLG) saliva. Also, we studied the presence of EGF in PG and SMG by immunohistochemistry. The mean (geometric) concentrations of EGF in PG saliva (2704 pg/ml, +/- SEM interval 2393-3056 pg/ml, n = 20) was higher (p less than 0.001) than in whole saliva (864 pg/ml, +/- 733-1019 pg/ml, n = 29), which in turn was higher (p less than 0.001) than in mixed SMG + SLG saliva (357 pg/ml, +/- 296-430 pg/ml, n = 16). No sex difference existed in any salivary gland EGF. Immunohistochemistry revealed EGF in the acinar cells of both PG and SMG, but only in PG there were prominent EGF deposits in luminal spaces. Our data suggest that EGF is produced by both PG and SMG, but that more of it is secreted from the PG. This result is new and challenges the general view that human salivary EGF is mainly from SMG. In mouse almost all salivary EGF comes from SMG and its amount is androgen dependent. Thus there are great differences in sources and regulation of salivary EGF between man and mouse.

Adult↗
PubMed1988

Production of monoclonal antibodies against murine dental papilla.

The mesenchymal component of the embryonic tooth, the dental papilla, shows unique cellular behavior. Only the dental papilla cells are capable of differentiating into odontoblasts. The dental papilla also directs the morphogenesis of the tooth. Our aim is to develop suitable markers for further studies on the molecular mechanisms behind the determination and differentiation of the dental mesenchymal cells. We have produced monoclonal antibodies against this embryonal cell population. Altogether 1114 enzymatically separated mesenchymes from dissected molar tooth germs of 17-day-old mouse embryos were fixed in paraformaldehyde (PFA) and sonicated in saline. A rat was immunized and hybridomas were produced by standard methods. The supernatants were screened by immunohistology, using both frozen sections and sections of PFA-fixed paraffin-embedded mouse molar teeth. Altogether, 19 wells produced antibodies reacting with dental tissues. One monoclonal antibody recognizes a 67,000 dalton intracellular antigen enriched in secretory odontoblasts and ameloblasts. The antigen is also found in osteoblasts and chondrocytes in the developing jaw, and in some cultured murine cells. The method described here appears to be successful for generating and screening monoclonal antibodies against the dental papilla.

Ameloblasts↗
PubMed1987

An autoradiographic study on the effect of epidermal growth factor on cell proliferation in erupting mouse incisors.

Epidermal growth factor (EGF) is a small polypeptide that induces precocious eyelid opening and incisor eruption in newborn mice; it stimulates cell proliferation in various cell types in vitro and in vivo. EGF was injected twice daily into newborn mice (0.4 microgram/g) and tritiated thymidine was injected subcutaneously 6 h before killing the mice. Longitudinal sections of the lower incisors showed significantly more thymidine-labelled mitoses in the pre-odontoblast and pre-ameloblast layers at the basal ends of incisors in EGF-treated mice than in control mice. Although these results indicate that the EGF-induced precocious tooth emergence is associated with a stimulation of cell proliferation in the root sheath, this tissue may not be the target for the actions of EGF. Earlier studies have shown that root growth is not a factor in the generation of the eruptive force and, as neither the pre-ameloblasts nor pre-odontoblasts express EGF-receptors, the stimulation of cell proliferation in the root-sheath region appears to be a result and not a cause of accelerated tooth eruption, i.e. the increase in root growth may meet the need to maintain the tooth positionally during the accelerated eruptive process.

Animals↗
PubMed1987

Localization and quantitation of 125I-epidermal growth factor binding in mouse embryonic tooth and other embryonic tissues at different developmental stages.

We have shown earlier that epidermal growth factor (EGF) inhibits morphogenesis and cell differentiation in mouse embryonic teeth in organ culture. This inhibition depends on the stage of tooth development so that only teeth at early developmental stages respond to EGF (A-M. Partanen, P. Ekblom, and I. Thesleff (1985) Dev. Biol. 111, 84-94). We have now studied the quantity and pattern of EGF binding in teeth at various stages of development by incubating the dissected tooth germs with 125I-labeled EGF. Although the quantity of 125I-EGF binding per microgram DNA stays at the same level, localization of 125I-EGF binding by autoradiography reveals that the distribution of binding sites changes dramatically. In bud stage the epithelial tooth bud that is intruding into the underlying mesenchyme has binding sites for EGF, but the condensation of dental mesenchymal cells around the bud does not bind EGF. At the cap stage of development the dental mesenchyme binds EGF, but the dental epithelium shows no binding. This indicates that the dental mesenchyme is the primary target tissue for the inhibitory effect of EGF on tooth morphogenesis during early cap stage. During advanced morphogenesis the binding sites of EGF disappear also from the dental papilla mesenchyme, but the dental follicle which consists of condensed mesenchymal cells surrounding the tooth germ, binds EGF abundantly. We have also studied EGF binding during the development of other embryonic organs, kidney, salivary gland, lung, and skin, which are all formed by mesenchymal and epithelial components. The patterns of EGF binding in various tissues suggest that EGF may have a role in the organogenesis of epitheliomesenchymal organs as a stimulator of epithelial proliferation during initial epithelial bud formation and branching morphogenesis. The results of this study indicate that EGF stimulates or maintains proliferation of undifferentiated cells during embryonic development and that the expression of EGF receptors in different organs is not related to the age of the embryo, but is specific to the developmental stage of each organ.

Animals↗
PubMed1987

Tenascin is associated with chondrogenic and osteogenic differentiation in vivo and promotes chondrogenesis in vitro.

The tissue distribution of the extracellular matrix glycoprotein, tenascin, during cartilage and bone development in rodents has been investigated by immunohistochemistry. Tenascin was present in condensing mesenchyme of cartilage anlagen, but not in the surrounding mesenchyme. In fully differentiated cartilages, tenascin was only present in the perichondrium. In bones that form by endochondral ossification, tenascin reappeared around the osteogenic cells invading the cartilage model. Tenascin was also present in the condensing mesenchyme of developing bones that form by intramembranous ossification and later was present around the spicules of forming bone. Tenascin was absent from mature bone matrix but persisted on periosteal and endosteal surfaces. Immunofluorescent staining of wing bud cultures from chick embryos showed large amounts of tenascin in the forming cartilage nodules. Cultures grown on a substrate of tenascin produced more cartilage nodules than cultures grown on tissue culture plastic. Tenascin in the culture medium inhibited the attachment of wing bud cells to fibronectin-coated substrates. We propose that tenascin plays an important role in chondrogenesis by modulating fibronectin-cell interactions and causing cell rounding and condensation.

Animals↗
PubMed1987

Levels and patterns of 125I-labeled transferrin binding in mouse embryonic teeth and kidneys at various developmental stages.

The iron-transporting serum glycoprotein, transferrin, is necessary for the cell proliferation, morphogenesis, and differentiation of mouse embryonic teeth and kidneys in organ culture. The stimulatory effect of transferrin is mediated by the binding of transferrin to its specific cell-surface receptor and by receptor-mediated endocytosis. Since, in both teeth and kidneys, the requirement for and responsiveness to transferrin depend on the developmental stage of the organ, we studied the binding of transferrin at various stages of tooth and kidney development by incubating tissues with 125I-labeled transferrin. The amount of bound transferrin was determined by measuring the tissue-incorporated radioactivity, and the binding sites were localized by autoradiography. During tooth development in vitro, the requirement for exogenous transferrin is lost as the teeth proceed from the early cap stage to the bell stage. The level of transferrin binding was found to decrease simultaneously, and in bell-stage teeth, the transferrin receptors were concentrated in the areas of most active cell proliferation. In kidneys, the number of transferrin receptors was highest at the stage during which the undifferentiated kidney mesenchyme becomes responsive to transferrin. These receptors were located in both the ureter epithelium and the metanephric mesenchyme, and they dramatically decreased in number with advancing kidney differentiation. The results of the present study indicate that, during the embryonic development of teeth and kidneys, the amount and localization of transferrin binding are correlated with cell proliferation. The number of transferrin receptors is highest during the developmental stages when cell proliferation is most active, and decreases with advancing differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Transferrin and tooth morphogenesis: retention of transferrin by mouse embryonic teeth in organ culture.

Transferrin is the only serum protein that is required for the early morphogenesis of mouse embryonic teeth in organ culture. Transferrin is able to support tooth morphogenesis and dental cell differentiation by stimulating cell proliferation. Its role in this process is restricted exclusively to iron transport, which takes place by receptor-mediated endocytosis of iron-loaded transferrin. A lipophilic iron chelator, pyridoxal isonicotinoyl hydrazone (PIH), can replace transferrin and support tooth morphogenesis in organ culture. We studied the effects of these two iron transporters on cell proliferation in tooth germs during culture. We found that Fe-PIH and transferrin stimulate proliferation to a similar extent in early cap-stage teeth of 14-day mouse embryos, but have no effect on cell proliferation in bell-stage teeth of 16-day mouse embryos. Day-16 teeth undergo morphogenesis in unsupplemented chemically defined medium, whereas transferrin or Fe-PIH is needed for the morphogenesis of day-14 teeth. Although the need for exogenous iron-transport molecules is lost with advancing development, the level of mitotic activity is still fairly high in bell-stage teeth. The abundant binding of transferrin in areas of active cell proliferation in bell-stage teeth also suggests that transferrin is still needed and used for the transport of iron into proliferating cells. Transferrin is not degraded by the process of receptor-mediated endocytosis. After releasing iron into a cell, transferrin is returned to the extracellular space and is reused. We therefore studied whether the transferrin needed by bell-stage teeth could be adequately supplied by endogenous transferrin synthesized or stored in tissue explants.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗