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PubMed1996

Occurrence of amorphous and crystalline mineral deposits at the epithelial-mesenchymal interface of incisors in the calcium-loaded rat: implication of novel calcium binding domains.

BACKGROUND: In order to clarify the regulatory factors that promote precipitation of enamel crystals in mammalian tooth germs, possible calcium binding domains were visualized at the epithelial-mesenchymal interface of rat incisor teeth by means of electron microscopy and X-ray microanalysis. METHODS: Adult rats were loaded with calcium (30 mM Ca) by vascular perfusion and further loaded through fixation and dehydration in the presence of high doses of calcium. RESULTS: Electron microscopy of anhydrously prepared Epon sections of the calcium-loaded rat incisors revealed numerous electron-dense granular deposits, enriched with calcium and phosphorus, scattering in the fibrous mantle dentin matrix and the intercellular spaces of the inner enamel epithelium, but not in the pulp tissues including the odontoblastic cells layer. The electron-dense deposits were specific for the enamel-related portion and were never shown to occur in the cementum-related portion. Proceeding incisally, dense deposits in both the mantle dentin matrix and presecretory ameloblast layer gradually converted to fine needlelike figures resembling the early enamel crystallites. Mineral deposits in experimental rats disappeared concomitant with the onset of normal mineralization of mantle dentin. There was no spatial correlation between the dense deposits and either stippled material or matrix vesicles. CONCLUSIONS: These results indicate the presence of novel calcium-binding domains in the enamel-related portion of the epithelial-mesenchymal interface of rat incisors that form enamellike crystallites under calcium-loaded conditions. A contribution of these putative calcium-binding domains in the induction and spontaneous formation of enamel crystals is suggested.

Animals↗
PubMed1997

Development of tight junctions between odontoblasts in early dentinogenesis as revealed by freeze-fracture.

BACKGROUND: Mature odontoblasts possess junctional structures constituted by adherens, gap, and tight junctions. Although adherens and gap junctions appear early between odontoblasts, there is no information on the appearance and development of tight junctions between odontoblasts. In this study, we have examined freeze-fracture replicas of early dentinogenesis to study the development of tight junctions between odontoblasts and to determine whether these junctions are of zonular or macular type. METHODS: Upper first molar tooth germs of Wistar rats between 1 and 3 days old were fixed in buffered 4% glutaraldehyde/4% formaldehyde and subsequently cryoprotected with cacodylate-buffered glycerol. Freeze-fracture replicas were obtained in a Balzers 301 apparatus, and early stages of dentinogenesis were examined in a Jeol 100 CX II electron microscope. RESULTS: In the stage of early dentine matrix prior to mineralization, odontoblasts exhibit only gap junctions. With the progression of development, the distal plasma membranes of odontoblasts show numerous short tight junctions formed by fused particles and grooves. In the stage of advanced mineralization, branched and continuous rows of fused particles or grooves constitute tight junctions of the focal or macular type. CONCLUSIONS: The present study shows that tight junctions of focal or macular type appear on distal plasma membrane of early odontoblasts during differentiation. Formation of tight junctions indicates the establishment of a distal membrane domain and maturation of odontoblasts. These events occur as mantle dentine formation ceases and circumpulpar dentine formation begins.

Age Factors↗
PubMed1976

The binding of calcium within the Golgi saccules of the rat odontoblast.

Odontoblasts of developing rat molar teeth were treated with OsO4-pyroantimonate to ascertain the localization of calcium. In addition, some tooth germs were incubated in solutions which were intended to allow for the escape of diffusible ions prior to fixation in OsO4-pyroantimonate. In tissues treated directly with OsO4-pyroantimonate, antimonate reaction product was found chiefly in abacus bodies and secretory granules of the Golgi region and in secretory granules in the distal pole of the cell. Lesser amounts of reaction product were found in the extracellular space, mitochondria, nucleus and generally throughout the cell. Tissues pre treated to allow for the escape of diffusible ions showed reaction product, identified as containing calcium, only in the abacus bodies and secretory granules. These results are considered to reflect the binding of calcium within the Golgi apparatus of the odontoblast. Moreover, since it has been shown by others that the abacus bodies and secretory granules contain collagen precursor, it is suggested that the collagen precursor is being seeded with calcium within the Golgi apparatus and that this intracellular calcium binding will play a role in facilitating the major wave of extracellular mineralization of the dentin which is to follow.

Animals↗
PubMed1992

Tubule formation and elemental detection in developing opossum enamel.

Most marsupials and some placental mammals possess enamel characterized by the presence of tubules, and the cellular origin of these structures has been the subject of a number of previous studies (See, for example, Lester, 1970; Azevedo and Goldberg, 1987). In the present report, tooth germs of the American opossum were examined to determine the structure and composition of enamel tubules during development and to analyze the enamel matrix relative to that of placental mammals with atubular enamel. For this purpose, tissues prepared by aqueous (decalcified and undecalcified) and anhydrous (undecalcified) methods were investigated by conventional transmission (TEM) and high voltage electron microscopy (HVEM), as well as by electron probe x-ray microanalysis (EPMA), selected-area electron diffraction (SAED), and electron spectroscopic imaging (ESI). Results indicate that most enamel tubules in the opossum begin as cytoplasmic remnants of Tomes' processes of ameloblasts. During development of the matrix, some of the tubules do not appear to be continuous throughout the prismatic layer. Sulfur is detectable around the lumen of the tubule in decalcified sections by EPMA and in and around the tubule by ESI. Calcium/phosphorus (Ca/P) molar ratios of the mineralizing matrix are generally higher than those found in enamel of other mammals and appear to decrease rather than increase with enamel maturation. The summary of data indicates the presence of sulfated glycoproteins or proteoglycans in this tissue, specifically around enamel tubules. Calcium and phosphorus are also present within the tubules, with the sulfated groups possibly binding calcium to prevent mineralization of the enamel tubules themselves.

Ameloblasts↗
PubMed2004

Tight junctions in differentiating ameloblasts and odontoblasts differentially express ZO-1, occludin, and claudin-1 in early odontogenesis of rat molars.

Little is known about the expression of associated proteins during the assembly of tight junctions (TJs). We studied the distribution of ZO-1, occludin, and claudin-1 between differentiating ameloblasts and odontoblasts in molar tooth germs from 1- to 3-day-old rats by confocal laser scanning microscopy. Immunoreactivity for ZO-1 was strong at proximal and distal junctional complexes of differentiating ameloblasts, while it was weak and punctuate at the distal region of differentiating odontoblasts. Occludin was immunoreactive at distal and proximal complexes of early differentiating ameloblasts and at distal regions of differentiating odontoblasts. However, in more advanced stages, occludin was only evident at the proximal complex of ameloblasts. Claudin-1 was strongly detected at the proximal complex but it was weak at distal complex of late differentiating ameloblasts. Thus, our results showed that ZO-1, occludin, and claudin-1 are differentially expressed as TJs assemble for regulating polarity and/or paracellular permeability in differentiating ameloblasts and odontoblasts.

Ameloblasts↗
PubMed1977

Tooth replacement in the teleost fish Prionurus microlepidotus Lacépède.

Previous studies on tooth replacement in lower vertebrates have been plagued by a lack of common integrative approaches and methods making it impossible to furnish a phylogenetic synthesis. This study is based on serial sections of the jaw of Prionurus microlepidotus. Each Toothgerm was characterized by its developmental stage and its position in the jaw. The relationship between the developmental stage of toothgerm and position in the jaw has been studied and expressed in several graphical illustrations. The following conclusions have been made: (1) The initiation of toothgerms in P. microlepidotus is governed by two Zahnreihen, which respectively initiate toothgerms on the lingual and labial side of the functioning teeth in an alternating pattern. (2) Therefore, functioning teeth in one locus are supplied by the alternate eruption of lingual and lubial toothgerms. (3) Advancing of tooth replacement in each locus is independent of functioning teeth and their successors in adjacent loci. (4) The disorders of replacement patterns are caused by an alternated rate of eruption of successive toothgerms as a response to unusual shedding of the functioning teeth.

Animals↗
PubMed1988

Morphological changes in the oral (buccopharyngeal) membrane in urodelan embryos: development of the mouth opening.

The ultrastructure of the oral (buccopharyngeal) membrane in the embryo of the urodelan, Hynobius tokyoensis, was examined by transmission (TEM) and scanning electron microscopy (SEM). The oral membrane consists of the stomodeal ectoderm and foregut endoderm, and is three to five cell layers thick at stage 24. The oral membrane gradually thickens as development proceeds. The stomodeal collar, derived from the ectoderm, is folded inward along the foregut endoderm. Tooth germs are formed partly by cells of the stomodeal collar and partly by mesenchymal cells and calcification takes place before hatching. Secretory granules, which are markers of epithelial differentiation, appear in some cells of the foregut endoderm. Within the oral membrane, the cells of the stomodeal collar become the basal cells, and the endodermal cells of the foregut become the apical cells of the future oral epithelium. Gaps are formed by the epithelial differentiation of the endodermal cells of the foregut in the oral membrane. The gaps connect with each other, with the stomodeum, and with the foregut. As a result of these events, the mouth opens at stage 43, just after hatching.

Animals↗
PubMed1997

A mammalian bicistronic transcript encoding two dentin-specific proteins.

We previously reported that the cDNA for dentin phosphophoryn, a non-collagenous protein involved in dentin mineralization, was located immediately downstream from the 3' end of dentin sialoprotein, another functionally related non-collagenous dentin protein (Ritchie, H. H., and Wang, L-H. (1996) J. Biol. Chem. 269, 3689-3702). We now demonstrate unequivocally that at the genomic level in mammal, these two functionally related proteins are indeed transcribed as one transcriptional unit suggesting that their combined presence at specific mineralization sites in tooth germ is mandatory if dentin mineralization is to proceed.

Amino Acid Sequence↗
PubMed1981

Palate morphogenesis. VI. Identification of stellate cells in culture.

Mesenchymal cells from the palate of mouse embryos at day 14.5 of gestation produce a minor population of stellate cells in culture. These cells are often bipolar and spindle-shaped with long cytoplasmic processes similar to neural-crest cells. Culturing of explants of palatal mesenchyme enriched for this type of cell. Stellate cells were the first to migrate from explants, followed by fibroblast-like cells and then by squamous cells. The majority of the cells in the explant were fibroblast-like. Squamous cells were present mostly in the anterior and mid-palate and least frequently in those from the posterior palate. They may represent tooth-germ epithelium. When pieces of palate were dissected out and cultured for enrichment of non-muscle contractile systems, most of the migrating cells were stellate. These may represent the highly migratory cells that are, in part, responsible for elevation of the palate shelf. Serotonin was measured in cultured mesenchymal cells from the palate. Its occurrence is consistent with regulation of movement of palate cells.

Animals↗
PubMed1991

Immunochemical and immunohistochemical studies, using antisera against porcine 25 kDa amelogenin, 89 kDa enamelin and the 13-17 kDa nonamelogenins, on immature enamel of the pig and rat.

Enamel proteins were extracted from the newly formed layer of immature porcine enamel, and the 25 kDa amelogenin, 89 kDa enamelin and 13-17 kDa nonamelogenins were purified. Specific antisera were raised against these proteins. Antibodies specific to the C-terminal region (residues 149-173) of the 25 kDa amelogenin were generated by absorption of the anti-25 kDa amelogenin serum with 20 kDa amelogenin, which contains residues 1-148 of the antigen. Immunoelectro-transfer blotting of the extracted porcine enamel proteins showed that the anti-25 kDa amelogenin serum recognized the 25 kDa and other low and high molecular weight amelogenins. The C-terminal specific anti-25 kDa amelogenin serum reacted only with amelogenins having molecular weights over 23 kDa. The anti-89 kDa enamelin serum recognized the 89 kDa enamelin and lower molecular weight proteins, but neither the amelogenins nor the 13-17 kDa nonamelogenins. The antiserum against the 13-17 kDa nonamelogenins showed no cross reactivity to the 89 kDa enamelin, but recognized higher molecular weight nonamelogenins. In immunohistochemical preparations of the porcine tooth germs, the 25 kDa amelogenin-like immunoreactivity over immature enamel decreased in a gradient from the enamel surface to the middle layer. In the inner layer immunoreactivity was concentrated over the prism sheaths. The C-terminal specific 25 kDa amelogenin-like immunoreactivity was intense at the outer layer of immature enamel and decreased sharply toward the middle layer. Prism sheaths were intensely stained by the antiserum to the 13-17 kDa nonamelogenins.(ABSTRACT TRUNCATED AT 250 WORDS)

Amelogenin↗
PubMed1989

[[Klippel-Trenaunay syndrome--an embryonic developmental disorder. A case report with reference to the orthodontic symptoms].

The Klippel-Trenaunay syndrome is a disturbance in the development of the mesodermal and ectodermal tissues occurring in utero. It is characterized by the clinical triad of unilateral soft tissue and bone hypertrophy, varicosities and hemangiomas. For dentists the orofacial manifestations of the syndrome are interesting: unilateral hypertrophy of the maxilla and mandible, mandibular joint and alveolar processes and tooth germs. In addition, the gingival margins are thickened, eruption is premature and there is marked facial asymmetry.

Angiomatosis↗