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J V Ruch

Publications and source records attributed to J V Ruch.

At least 37 records · Page 2Linked to original sources
PubMed1998

Expression and localization of laminin-5 subunits during mouse tooth development.

Tooth morphogenesis is regulated by epithelial-mesenchymal interactions mediated by the basement membrane (BM). Laminins are major glycoprotein components of the BMs, which are involved in several cellular activities. The expression and localization of the alpha3, beta3, and gamma2 laminin-5 subunits have been analyzed by in situ hybridization and immunohistochemistry during mouse molar development. Initially (E12), mRNAs of all subunits were detected in the entire dental epithelium and the corresponding proteins were located in the BM. During cap formation (E13-14), transcripts for the alpha3 and gamma2 subunits were localized in the outer dental epithelium (ODE), whereas the beta3 subunit mRNA was present in the inner dental epithelium (IDE). During the early bell stage (E16), immunoreactivity for all subunits disappeared from the BM along the IDE, although intense signals for beta3 mRNA were detectable in cells of the IDE. Subsequently, when the dentinal matrix was secreted by odontoblasts (E18-19.5), mRNAs of all three subunits were re-expressed by ameloblasts, and the corresponding proteins were detected in ameloblasts and in the enamel matrix. Tissue recombination experiments demonstrated that when E16 IDE or ODE was associated with E18 dental papilla mesenchyme, immunostaining for all laminin-5 subunits disappeared from the BM, whereas when cultured with non-dental limb bud mesenchyme, they remained positive after 48 hr of culture. These results suggest that the temporospatial expression of laminin-5 subunits in tooth development, which appears to be differentially controlled by the dental mesenchyme, might be related to the enamel organ histo-morphogenesis and the ameloblast differentiation.

Ameloblasts↗
PubMed1998

Expression and localization of laminin-5 subunits in the mouse incisor.

Laminin-5 is associated with several epithelial tissues and forms part of the anchoring filaments of hemidesmosomes. Recent data have shown that the expression of laminin-5 subunits is impaired in junctional epidermolysis bullosa (JEB), and, in these patients, enamel hypoplasia is commonly observed. Rodent incisors are continuously growing teeth with an asymmetry between their labial and lingual sides. Enamel matrix formation is restricted to the labial side. We have analyzed the changes in the expression and localization of laminin-5 subunits (alpha3, beta3, and gamma2) in lower incisors of the mouse. The apical loop located at the end of the labial side contained stem cells and showed expression for all laminin-5 subunits. In the anterior direction, the inner dental epithelial cells (IDE) transiently lost the immunoreactivity for all subunits, whereas the transcripts for the beta3 subunit remained in the IDE. All subunit mRNAs and proteins were expressed in ameloblasts facing predentine and also in secretory and maturation stage ameloblasts. Enamel matrix contained laminin-5. On the lingual side, the expression of laminin-5 subunits was continuous from the epithelial root sheath to the epithelial rests of Malassez in the periodontal ligament. These results suggest that spatial and temporal regulation of laminin-5 subunits correlates with the histogenesis of the dental organ, ameloblast differentiation, and enamel formation and also that laminin-5 plays a role in the adhesion between dental epithelial cells and the extracellular matrix (enamel or dentine) in areas where the dental basement membrane is absent.

Amelogenesis↗
PubMed1998

Correlation between apoptosis distribution and BMP-2 and BMP-4 expression in vestigial tooth primordia in mice.

The eutherian dental formula consists of three incisors, one canine, four premolars and three molars in each dental quadrant. Muroid evolution led to a reduction in the number of teeth, with one incisor separated from three molars by a long diastema. However, seven vestigial tooth primordia (D1-5, R1-2) have been detected in the embryonic diastemal area of the mouse maxilla between embryonic days (ED) 12.5 and 13.5. Computer-aided 3D reconstructions were used to analyse the temporo-spatial pattern of apoptosis during regression in the two largest and most distal vestiges (R1, R2). These structures have been widely considered as the primordium of the first upper molar and, accordingly, related molecular data have been interpreted exclusively in terms of progressive molar development. The spatial distribution of epithelial apoptosis, which affected the R1 and R2 rudiments in two consecutive waves on ED 12.5 and 13.5, respectively, was compared with our earlier data on expression of genes encoding bone morphogenetic proteins (BMP-2 and BMP-4). Similar temporo-spatial patterns of apoptosis and expression of BMP, specifically confined to the epithelium of the rudimentary tooth primordia, strongly support involvement of BMPs in the regulation of epithelial apoptosis during odontogenesis.

Animals↗
PubMed1998

Localization of antigens associated with adherens junctions, desmosomes, and hemidesmosomes during murine molar morphogenesis.

Epitheliomesenchymal interactions are known to play a crucial role during odontogenesis. Since epithelial cell-cell and cell-matrix interactions may also be involved in enamel organ histomorphogenesis, we investigated the localization of proteins associated with junctional complexes in mouse and rat first lower molars by indirect immunofluorescence. Adherens junctions were detected using antibodies directed against E-cadherin, beta-catenin, and plakoglobin (gamma-catenin). Desmosomes were localized with antibodies against desmoglein, and hemidesmosomes using antibodies against BP-230 and HD-1 proteins. When the inner dental epithelium differentiates, a decrease of E-cadherin, plakoglobin, and BP-230 is seen. An asymmetric distribution of plakoglobin, desmoglein, and BP-230 between the lateral and medial side of the tooth exists; desmoglein, which was first restricted to the gubernaculum dentis, progressively accumulated in the stellate reticulum, the stratum intermedium, and the basal pole of ameloblasts. The specific temporospatial distributions patterns of these antigens suggests a direct involvement of adherens junctions, desmosomes, and hemidesmosomes in the development of the murine first lower molar.

Animals↗
PubMed1998

Early stages of tooth morphogenesis in mouse analyzed by 3D reconstructions.

Computer-aided 3D reconstructions were used to investigate early odontogenesis in the ICR mouse, from the dental lamina to the cap stage. The diastemal region of the maxilla was not an empty zone: five transient epithelial rudiments (D1-D5) were found between ED 12.5-13.5. Two further rudiments (R1 and R2) were observed between D5 and the maxillary first molar primordium, whose bud emerged at ED 13.5. These rudiments might be related to vestiges of ancestral teeth. During this period, only an epithelial lamina was observed in front of the bud-shaped molar epithelium in the cheek region of the mandible. Apoptosis plays an important role in the reduction of antemolar rudiments in the maxilla and in the remodeling of the epithelium anterior to the M1 bud and cap in both jaws: two successive waves of apoptosis were detected in the mandible and in the maxilla. Computer-aided 3D reconstructions clearly demonstrated that morphologically different developmental stages coexist along the anteroposterior axis of M1 in both jaws.

Animals↗
PubMed1998

Expression of p21(WAF1/CIP1) during mouse odontogenesis.

p21(WAF1/CIP1) is a cyclin-dependent kinase (Cdk) inhibitor. This protein may function during development as an inducible growth inhibitor that contributes to cell cycle exit and differentiation. The expression pattern of p21 during mouse embryogenesis was correlated with terminal differentiation of multiple cell lineages including skeletal muscles, cartilage, skin and nasal epithelium. p21 expression was analyzed by in situ hybridization during odontogenesis as well as during in vitro tooth development in chemically defined medium with or without retinoic acid. p21 transcripts were detected in the restricted area of the inner dental epithelium during late cap and initial bell stages and then confined to the post-mitotic odontoblasts and ameloblasts. The replicating cells were devoid of any signal. The distribution of p21 mRNA in vitro, whatever the culture conditions, was similar to the in vivo pattern. p21 protein immunolocalization was superimposed on the transcripts distribution but more restricted in ameloblasts. TGFbeta1 is known to induce p21 expression. During dental cytodifferentiations, TGFbeta1 and p21 expressions overlap. Growth inhibition by TGFbeta1 may be associated with p21 induction.

Ameloblasts↗
PubMed1998

Effects of aFGF, bFGF, TGFbeta1 and IGF-I on odontoblast differentiation in vitro.

In this work, we investigated the effects of aFGF and bFGF alone or combined with TGFbeta1 or IGF-I on odontoblast differentiation. Trypsin-isolated dental papillae from day 17 mandibular first molar were cultured in semisolid-agar medium for 6 d. Our results demonstrated that aFGF, bFGF or combinations of these promoted cell polarization at the periphery of the dental papillae. Moreover, simultaneous addition of aFGF and TGFbeta1 to dental papillae cultures induced both polarization and functional differentiation of odontoblast-like cells, as well as extracellular matrix deposition. Combination of aFGF or bFGF with IGF-I caused cell polarization at the surface of dental papillae, but matrix secretion was restricted to a few explants. In the presence of bFGF and TGFbeta1, the explants had pronounced cell elongations but no matrix deposition. These results indicate that aFGF or bFGF is not able to induce odontoblast differentiation alone. However, both aFGF and bFGF can act synergistically with TGFbeta1 and IGF-I to strengthen their inductive effects and promote gradients of cytological and functional changes in odontoblast-like cells.

Animals↗
PubMed1998

Activin and bone morphogenetic protein (BMP) signalling during tooth development.

Activins and bone morphogenetic proteins (BMPs) belong to the transforming growth factor-beta (TGF-beta) superfamily of structurally related cytokines. Results of recent in vivo and in vitro studies suggest that activin A, several BMPs, and their cell surface receptors may participate in tooth development. In addition, follistatin an extracellular protein that may interact with activin and BMP signaling, appears to be involved. This review focuses on recent advances in relation to the roles of activin/BMP receptors, their ligands and extracellular modifiers during tooth development.

Activin Receptors↗
PubMed1998

Comparative analysis of mouse DSP and DPP expression in odontoblasts, preameloblasts, and experimentally induced odontoblast-like cells.

Dentin sialoprotein (DSP) and dentin phosphoproteins (DPP) are uniquely expressed by differentiating and fully differentiated mature odontoblasts. It is likely that DSP and DPP actively participate in the conversion of predentin to dentin. To compare the expression patterns of DPP and DSP, we constructed mouse cDNA probes. Northern analyses confirmed their specific expressions in tooth germ-derived RNA and showed that the probes reacted with similar or identical multiple transcripts. In situ hybridization indicated that DSP and DPP transcripts were uniquely detected and codistributed in developing mouse odontoblasts and preameloblasts. These data, as well as the adjacent positioning of the DSP and DPP coding sequences, suggest that common regulatory mechanisms control DSP and DPP expressions. Dental papillae cultures, in which odontoblast differentiation was experimentally induced with TGFbeta1 combined with heparin, were used to show that the two molecules are also coexpressed under in vitro conditions.

Ameloblasts↗
PubMed1998

Odontoblast commitment and differentiation.

Histological and cytological organization confer specificity to the odontoblasts. These postmitotic, neural crest derived, polarized cells are aligned in a single layer at the periphery of the dental pulp and secrete the organic components of predentin-dentin. The developmental history of these cells demands a cascade of epigenetic signalling events comprising the acquisition of odontogenic potential by neural crest cells, their patterning in the developing jaws, the initiation of odontogenesis through interaction with the oral epithelium, commitment, and tooth-specific spatial distribution of competent preodontoblasts able to overtly differentiate. Recent experimental investigations are critically summarized, many open questions are stressed, and current hypotheses concerning the control of terminal odontoblast differentiation are outlined.

Animals↗
PubMed1997

Distribution of transforming growth factor beta1-binding proteins and low-affinity receptors during odontoblast differentiation in the mouse.

Transforming growth factor-beta1 (TGF-beta1) was immunolocalized within differentiated odontoblasts and ameloblasts while LAP-beta1 was detected at the apicol pole of odonotoblasts and ameloblasts and in predentine. Anti-LAP-beta1 antibodies also stained the epithelial-mesenchymal junction (EMJ). Decorin was immunolocalized in young functional odonotoblasts and in both predentine and dentine. Biglycan was similarly distributed but absent from dentine. Immunostaining with anti-latent TGF-beta1 binding protein-1 (LTBP-1) showed fibrillar structures located at the EMJ and between predontoblasts and odontoblasts; at older states staining was restricted to the dental papilla and sac. Thus differentiated odonotoblasts express TGF-beta1 and in a more restricted manner decorin, biglycan and LAP-beta1; it can be assumed that TGF-beta1 is able to interact with the three molecules present in predentine. Earlier, LTBP-1 and LAP-beta1, both present at the EMJ, may contribute to odontoblast differentiation.

Activin Receptors, Type I↗
PubMed1997

The activin-binding protein follistatin is expressed in developing murine molar and induces odontoblast-like cell differentiation in vitro.

It has recently been shown that mice deficient in activin-beta A subunits and follistatin exhibit major defects in dentition. To increase understanding of the roles played by these molecules during tooth development, we determined the temporospatial expression of activin-beta A subunit and follistatin messenger RNA and their corresponding proteins in developing murine molars (between day E 14 and 2 days after birth). The effects of recombinant human activin A and its binding protein follistatin on odontoblast differentiation were also studied in cultures of dental papillae (DP) isolated from the mandibular first molars of E-17-day mice. In situ hybridization indicated that transcripts for activin-beta A subunit were abundant in pre-odontoblasts at the tips of forming cusps prior to odontoblast terminal differentiation, and transcripts for follistatin in overlying inner enamel epithelial cells (pre-ameloblasts). Pre-odontoblasts were also weakly immunoreactive in relation to activin-beta A subunit, pre-ameloblasts in relation to follistatin. When follistatin was added at different concentrations to a DP culture model (2-14 nmol/DP) together with heparin at constant concentration, differentiation of odontoblast-like cells was induced, as evidenced by polarization and deposition of extracellular matrix in vitro, to extents depending on the follistatin concentration. In contrast, the addition of activin A (2 nmol/DP) had no effect on the differentiation parameters studied. These findings suggest that the activin-follistatin system regulates odontoblast differentiation during tooth development. In particular, we suggest that binding of endogenous activin A by follistatin may allow odontoblast terminal differentiation to occur.

Activins↗
PubMed1997

Mouse molar morphogenesis revisited by three-dimensional reconstruction. III. Spatial distribution of mitoses and apoptoses up to bell-staged first lower molar teeth.

Computer-assisted 3D reconstructions were used to follow the development of the embryonic mouse first lower molar (M1). At ED 12.5, the thickening of the oral epithelium, which was thought to correspond to the molar dental lamina, regressed in its anterior part as a result of apoptosis. Only the posterior part later gave rise to molars. The transition to the cap stage entailed medial and lateral extensions of the dental epithelium. The growth and histo-morphogenesis of the enamel organ as well as cervical loop formation proceeded more rapidly in the anterior part of the M1 during the cap and early bell stages producing significant morphological differences along the antero-posterior axis. Apoptosis was temporarily intensive in the anterior part of the bud- and cap-shaped epithelium and thus pointed domains which do not participate in the formation of the final M1 enamel organ. In the well-formed cap, apoptoses displayed maximum concentration in the enamel knot (EK). No increase in the number of metaphases could be detected in the vicinity of the EK. Mitoses were distributed throughout the epithelial compartment until cap stage and then mainly concentrated in the inner dental epithelium at the early bell stage. At this later stage, either lateral views or thick virtual sections performed in the reconstruction demonstrated a clear cut distribution of mitoses and apoptoses in the enamel organ. At the early bell stage, mitoses in the mesenchyme demonstrated an increasing postero-anterior gradient.

Animals↗
PubMed1996

Apoptosis is involved in the disappearance of the diastemal dental primordia in mouse embryo.

Three transient dental primordia (D1, D2 and D3) exist in the upper diastema in mouse embryos and their regression is associated with the presence of cell death. In order to specify the type of cell death and its temporo-spatial distribution, staining with hematoxylin, supravital staining with Nile Blue, TUNEL method, electron microscopic analysis and computer assisted 3-D reconstructions were performed. These data demonstrated that apoptosis is involved in the disappearance of the diastemal dental rudiments. Apoptosis occurred first with prevalence in the buccal part of the epithelium of the diastemal dental primordia and extended later to the whole epithelium of the dental rudiments and the dental lamina interconnecting them with the incisor and molar epithelia. Cell death occurred only sporadically in the adjacent mesenchyme. The prospective upper diastema in mouse embryos may provide a model for studies of developmental determination of toothless areas in the jaw as well as a tool for analyses of regulatory mechanisms of programmed cell death in morphogenesis.

Animals↗
PubMed1996

Mouse molar morphogenesis revisited by three dimensional reconstruction. I. Analysis of initial stages of the first upper molar development revealed two transient buds.

Early stages of tooth development in the maxillary cheek region in the mouse were investigated by combined analysis of histological sections, computer assisted 3D reconstructions and morphometry. In ED 12.5 embryos, 3D reconstructions revealed an accessory epithelial bud (R1) and a large bud (R2), which appeared as a single bud-shaped epithelium in frontal sections. This developmentally most advanced dental epithelium in the mouse embryonic maxilla until ED 13.5, generally considered as the bud of the first molar, regressed during later development. Meanwhile the bud and cap of the first upper molar originated more posteriorly, from ED 13.5. The regression of R1 and R2 was associated with epithelial apoptosis. Apoptotic cells and bodies were apparent on sections in the R1 epithelium from ED 12.5. The R2 epithelium maintained the large bud-shaped appearance on sections, representing the largest part of the dental epithelium in the maxillary cheek region until ED 14.0; apoptoses were detected there as late as from ED 13.5. During regression, the R2 rudiment was transformed into the medial and lateral epithelial ridges, posteriorly in continuity with the arising cap of the first molar. The reduced R1 epithelium seemed to contribute to the medial ridge. These results should be taken into consideration in the interpretation of early odontogenesis in the upper jaw in the mouse. The interesting problem of the identification of tooth homology of the rudiments should be elucidated by further comparative morphological and paleontological investigations.

Animals↗
PubMed1996

Mouse molar morphogenesis revisited by three-dimensional reconstruction. II. Spatial distribution of mitoses and apoptosis in cap to bell staged first and second upper molar teeth.

Tooth morphogenesis is a complex multifactorial process in which differential mitotic activities and cell death play important roles. Upper first (m1) and second (m2) molars from mouse embryos were investigated from early cap to bell stage. m2 differed from m1 by delayed origin of the enamel grooves delimiting the protrusion of the cap bottom towards the dental papilla, and retardation of the enamel knot formation. The width of the m2 enamel organ was conspicuously smaller during cap formation and length remained smaller throughout the period of observation. Formation of the cap depression was comparable in m1 and m2, however margins delimiting the enamel organ cavity arose in m1 and m2 as mirror images. Attempts were made to correlate changes in the distribution of apoptotic cells and bodies and/or mitoses with morphogenesis. These cellular activities were recorded from histological sections and represented in space using computer-assisted three-dimensional reconstructions. Mitoses in the epithelial compartment were associated with the development of the cervical loop. In the mesenchyme of m1 at early bell stage, a postero-anterior increasing gradient of mitoses was observed which might be correlated with the anterior growth of the molar. Cells in the enamel knot demonstrated a high level of apoptosis, retarded in m2, but absolutely no division. Apoptotic processes were also involved in the anterior delimitation of the m1 epithelium. Apoptosis might correspond to the programmed destruction of cells whose function had to be suppressed or whose potential activity had to be avoided.

Animals↗