RESUMEN
Saethre-Chotzen syndrome (acrocephalo-syndactyly type III, ACS III) is an autosomal dominant craniosynostosis with brachydactyly, soft tissue syndactyly and facial dysmorphism including ptosis, facial asymmetry and prominent ear crura. ACS III has been mapped to chromosome 7p21-22. Of interest, TWIST, the human counterpart of the murine Twist gene, has been localized on chromosome 7p21 as well. The Twist gene product is a transcription factor containing a basic helix-loop-helix (b-HLH) domain, required in head mesenchyme for cranial neural tube morphogenesis in mice. The co-localisation of ACS III and TWIST prompted us to screen ACS III patients for TWIST gene mutations especially as mice heterozygous for Twist null mutations displayed skull defects and duplication of hind leg digits. Here, we report 21-bp insertions and nonsense mutations of the TWIST gene (S127X, E130X) in seven ACS III probands and describe impairment of head mesenchyme induction by TWIST as a novel pathophysiological mechanism in human craniosynostoses.
Asunto(s)
Acrocefalosindactilia/genética , Mutación , Proteínas Nucleares/genética , Factores de Transcripción/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromosomas Humanos Par 7 , ADN , Femenino , Secuencias Hélice-Asa-Hélice , Humanos , Masculino , Ratones , Datos de Secuencia Molecular , Mutagénesis Insercional , Linaje , Fenotipo , Polimorfismo Conformacional Retorcido-Simple , Proteína 1 Relacionada con TwistRESUMEN
The expression of type IV collagen mRNA during mouse intestinal morphogenesis was examined by in situ hybridization using a cDNA probe corresponding to mRNA for alpha 1 (IV) chain. Type IV collagen mRNA is detected in the embryonic mesenchymal cells at early stages of development (12 d of gestation). A segregation of mesenchymal cells expressing high levels of type IV collagen mRNA in close vicinity of the epithelium occurs just before villus formation. During villus outgrowth, type IV collagen mRNA, still confined to mesenchyme-derived tissues, is progressively restricted to the mucosal connective tissue (the lamina propria) and to a lesser extent to the muscular layers. In the adult, the amount of messenger is quite low as compared to the level found in the developing intestine and the in situ hybridization signal, indistinguishable from the background, is uniform throughout the whole intestinal wall. At all developmental stages no detectable specific hybridization signal is virtually observed over the epithelium cell layer. These results show that high amounts of the type IV collagen messenger are detected during phases of intensive morphogenetic events. Furthermore, they reinforce the notion already gained previously (Simon-Assmann et al. 1988) that the mesenchymal compartment is the principal endogenous source of type IV collagen. They also indicate that the continuous migration of epithelial cells along the basement membrane of intestinal villi in the mature organ is not accompanied by a significant remodeling of the collagen IV network.
Asunto(s)
Colágeno/genética , Intestinos/embriología , Músculo Liso/embriología , ARN Mensajero/genética , Envejecimiento , Animales , Northern Blotting , Sondas de ADN , Intestinos/crecimiento & desarrollo , Sustancias Macromoleculares , Ratones , Ratones Endogámicos , Morfogénesis , Desarrollo de Músculos , Músculo Liso/crecimiento & desarrollo , Hibridación de Ácido Nucleico , ARN Mensajero/metabolismoRESUMEN
OBJECTIVE: The aim of this study was to improve the strength and quality of the titanium-hydroxyapatite interface in order to prevent long-term failure of the implanted devices originating from coating delamination and to test it in an in-vivo model. METHODS: Ti disks and dental commercial implants were etched in Kroll solution. Thermochemical treatments of the acid-etched titanium were combined with sol-gel hydroxyapatite (HA) coating processes to obtain a nanoporous hydroxyapatite/sodium titanate bilayer. The sodium titanate layer was created by incorporating sodium ions onto the Ti surface during a NaOH alkaline treatment and stabilized using a heat treatment. HA layer was added by dip-coating in a sol-gel solution. The bioactivity was assessed in vitro with murine MC3T3-E1 and human SaOs-2 cells. Functional and histopathological evaluations of the coated Ti implants were performed at 22, 34 and 60days of implantation in a dog lower mandible model. RESULTS: Nanoporous hydroxyapatite/sodium titanate bilayer on titanium implants was sensitive neither to crack propagation nor to layer delamination. The in vitro results on murine MC3T3-E1 and human SaOs-2 cells confirm the advantage of this coating regarding the capacity of cell growth and differentiation. Signs of progressive bone incorporation, such as cancellous bone formed in contact with the implant over the existing compact bone, were notable as early as day 22. Overall, osteoconduction and osteointegration mean scores were higher for test implants compared to the controls at 22 and 34 days. SIGNIFICANCE: Nanoporous hydroxyapatite/sodium titanate bilayer improves the in-vivo osteoconduction and osteointegration. It prevents the delamination during the screwing and it could increase HA-coated dental implant stability without adhesive failures. The combination of thermochemical treatments with dip coating is a low-cost strategy.
Asunto(s)
Implantes Dentales , Durapatita , Óxidos , Titanio , Animales , Materiales Biocompatibles Revestidos , Perros , Humanos , Ratones , Microscopía Electrónica de Rastreo , Nanoporos , Oseointegración , Propiedades de SuperficieRESUMEN
Comparing the sequences of the murine Brachyury T gene protein with the NKFB, rel and dorsal protein family reveals weak sequence similarities, including a DNA-binding/dimerization motif. These similarities may reflect homologies between these proteins or may only be of phylogenetic relevance.
Asunto(s)
Proteínas Fetales/genética , FN-kappa B/genética , Proteínas Tirosina Quinasas/metabolismo , Proteínas Proto-Oncogénicas/genética , Proteínas de Dominio T Box , Secuencia de Aminoácidos , Datos de Secuencia Molecular , Filogenia , Proteínas Proto-Oncogénicas c-rel , Homología de Secuencia de AminoácidoRESUMEN
M-twist is the murine homolog of the Drosophila twist gene which is a zygotic target for maternal genes that establish embryonic dorso-ventral polarity and is necessary for mesoderm formation. We recently showed that before gastrulation, M-twist transcripts are detected in morulae and blastocysts, then in extra-embryonic tissues of early implanted mouse embryos before the onset of gastrulation, and we suggested that M-twist might be involved in embryonic polarity (Stoetzel et al., submitted). Here, using in situ hybridization on whole mount embryos, we present the expression pattern of M-twist from primitive streak stage up to 10.5 days p.c. In implanted embryos, M-twist is first expressed in extra-embryonic tissues, then in embryo proper around egg cylinder stage within some embryonic ectodermal cells of the primitive streak. Slightly later, scattered cells within the amniotic cavity apparently detached from the primitive streak also express the gene. Then, M-twist transcripts accumulate in head mesenchyme, the first aortic arches, somites and lateral mesoderm and, as development proceeds, successively the second, third and fourth branchial arches, the anterior limb buds and, finally, the posterior limb buds. Thus M-twist expression in implanted embryos occurs first along a dorso-ventral gradient pattern until the headfold stage, then it is gradually observed along the rostro-caudal axis of the embryos as development procedes in the mesodermal cell layer and in neural crest cell derivatives. In addition, we show the existence of some previously undescribed subsets of scattered cells that express M-twist and thus might participate in murine embryo development.
Asunto(s)
Desarrollo Embrionario y Fetal/genética , Regulación del Desarrollo de la Expresión Génica , Factores Reguladores Miogénicos , Proteínas Nucleares/genética , Animales , Secuencia de Bases , Cartilla de ADN/genética , Drosophila/embriología , Drosophila/genética , Implantación del Embrión , Femenino , Edad Gestacional , Hibridación in Situ , Ratones , Datos de Secuencia Molecular , Embarazo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteína 1 Relacionada con TwistRESUMEN
Recent developments in tooth-tissue engineering require that we understand the regulatory processes to be preserved to achieve histomorphogenesis and cell differentiation, especially for enamel tissue engineering. Using mouse first lower molars, our objectives were: (1) to determine whether the cap-stage dental mesenchyme can control dental epithelial histogenesis, (2) to test the role of the primary enamel knot (PEK) in specifying the potentialities of the dental mesenchyme, and (3) to evaluate the importance of positional information in epithelial cells. After tissue dissociation, the dental epithelium was further dissociated into individual cells, re-associated with dental mesenchyme, and cultured. Epithelial cells showed a high plasticity: Despite a complete loss of positional information, they rapidly underwent typical dental epithelial histogenesis. This was stimulated by the mesenchyme. Experiments performed at E13 demonstrated that the initial potentialities of the mesenchyme are not specified by the PEK. Positional information of dental epithelial cells does not require the memorization of their history.
Asunto(s)
Morfogénesis/fisiología , Germen Dentario/embriología , Animales , Apoptosis/fisiología , Comunicación Celular , Diferenciación Celular , Órgano del Esmalte/embriología , Células Epiteliales/fisiología , Epitelio/embriología , Procesamiento de Imagen Asistido por Computador , Imagenología Tridimensional , Mesodermo/fisiología , Ratones , Ratones Endogámicos ICR , Técnicas de Cultivo de TejidosRESUMEN
Until recently, Bardet-Biedl syndrome was considered as a classic autosomal recessive condition. The disorder is defined by the association of the following clinical features: retinitis pigmentosa, polydactyly, obesity, hypogonadism, and possible mental retardation. This syndrome leads to multiple handicaps (visual impairment, complications of obesity, kidney failure, endocrine dysfunction). This condition, apparently clearly defined from a clinical point of view, appears to be genetically heterogenous. To date, six different genes have been identified: BBS1, BBS2, BBS4, BBS6, BBS7 and BBS8. Interestingly, this condition has recently been linked to a failure of cellular ciliogenesis. Moreover, this disorder is characterized by an additional degree of complexity, as it is the first example of triallelic inheritance described in human beings. However, this new finding appears to be less frequent than expected in this syndrome.
Asunto(s)
Síndrome de Bardet-Biedl/genética , Síndrome de Bardet-Biedl/diagnóstico , HumanosRESUMEN
Rodent incisors are continuously growing teeth and enamel deposition is restricted to the labial side. In the present study, the expression of laminin-5 subunits (alpha3, beta3 and gamma2) has been analyzed by in situ hybridization in developing mouse lower incisors and compared to that reported in the molar. At the bud stage (E12), mRNAs for all subunits were detected in the whole epithelial thickening. At E14, when histogenesis had started, transcripts for alpha3 and gamma2 subunits were restricted to the outer dental epithelium (ODE), whereas the beta3 subunit was intensely expressed in the inner dental epithelium (IDE). A transient expression for alpha3 subunit was seen in the enamel knot area and disappeared at E15. Subsequently, all laminin-5 subunit genes were re-expressed in differentiating ameloblasts on the labial side. Similar patterns of transcription were observed in incisor and molar, suggesting that the differential expression of laminin-5 subunits in the IDE might be involved in the histogenesis of the IDE and ameloblast differentiation. At E16.5, cells of the IDE at the anterior extremity of the incisor and in the anterior part of the lingual IDE expressed transcripts for alpha3 and beta3 but not for gamma2 subunit. Similar expression patterns were observed in the enamel-free areas of the E18 molar. This specific expression might thus be related to cells that do not differentiate as functional ameloblasts. Throughout incisor development, intense expression for all laminin-5 subunits was restricted to the labial side of the cervical loop. The asymmetrical expression of laminin-5 might be related to incisor morphogenesis and to the differences in histogenesis and cytodifferentiation of the IDE that exist in the labial versus lingual aspect of the cervical loop.
Asunto(s)
Moléculas de Adhesión Celular/biosíntesis , Incisivo/embriología , Diente Molar/embriología , Animales , Moléculas de Adhesión Celular/química , Epitelio/embriología , Epitelio/metabolismo , Inmunohistoquímica , Hibridación in Situ , Ratones , Modelos Biológicos , Factores de Tiempo , Transcripción Genética , KalininaRESUMEN
Early X-twi expression has been now investigated from egg laying to the early neurulation stages in Xenopus embryos, using both in situ hydridization and the more sensitive techniques of RT-PCR. We show that in unfertilized eggs, a decreasing gradient of X-twi transcript distribution is observed from animal to vegetative caps. X-twi RNA can be weakly detected at stages prior to gastrulation, and with increased intensity from stage 8 onwards. At blastula, X-twi transcripts are located towards the animal pole, and as gastrulation begins, they are detected in the developing axial mesoderm and then they accumulate in the sensorial layer of the neurectoderm, the mesodermal layer and in neural crest cells up to late neurula stages. We show, in addition, that in lithium-chloride- and UV-treated Xenopus embryos (that are respectively both "anteriorized/dorsalized" and in "posteriorized/ventralized"), X-twi RNA is detected in cells in similar positions to those that express X-twi in normal embryos. As a whole, our results show that X-twi is expressed even when regionalization of the mesoderm is disturbed and raises the question of a putative function of X-twi prior to gastrulation.
Asunto(s)
Blastocisto , Proteínas de Unión al ADN/genética , Ectodermo/química , Regulación del Desarrollo de la Expresión Génica/fisiología , Mesodermo/química , Proteínas de Xenopus , Animales , Tipificación del Cuerpo , Gástrula , Cloruro de Litio/farmacología , Óvulo/química , ARN Mensajero/análisis , Proteína 1 Relacionada con Twist , Rayos Ultravioleta , Xenopus laevisRESUMEN
Tissue engineering has merged with stem cell biotechnology with development of new sources of transplantable biomaterials for the treatment of bone tissue diseases. Bone defects are expected to benefit from this new biotechnology because of the low self-regenerating capacity of bone matrix secreting cells. The differentiation of stem cells to bone cells using bi-functionalized multilayered particles is presented. The functionalized particles are composed of poly-glutamic acid (PGA) and poly-L-lysine (PLL) with two bone growth factors (BMP-2 and TGFbeta1) embedded into the multilayered film. The induction of bone from these bioactive particles incubated with embryonic stem cells was demonstrated in vitro. We report the demonstration of a multilayered particle-based delivery system for inducing bone formation in vivo. This new strategy is an alternative approach for in vivo bone formation.
Asunto(s)
Sistemas de Liberación de Medicamentos , Péptidos y Proteínas de Señalización Intercelular/farmacología , Nanoestructuras/química , Osteogénesis/efectos de los fármacos , Osteogénesis/fisiología , Animales , Proteína Morfogenética Ósea 2/farmacología , Masculino , Ratones , Ratones Desnudos , Microscopía Confocal , Poliestirenos/química , Ingeniería de Tejidos , Factor de Crecimiento Transformador beta1/farmacologíaAsunto(s)
Cromosomas Humanos Par 2 , Cromosomas Humanos Par 7 , Proteínas Represoras , Factores de Transcripción/genética , Secuencia de Aminoácidos , Animales , Mapeo Cromosómico , Clonación Molecular , Secuencia Conservada , Drosophila/genética , Proteínas de Drosophila , Secuencias Hélice-Asa-Hélice , Humanos , Ratones , Datos de Secuencia Molecular , Proteínas Nucleares/química , Proteínas Nucleares/genética , Alineación de Secuencia , Factores de Transcripción/química , Proteína 1 Relacionada con TwistAsunto(s)
Blefarofimosis/genética , Cromosomas Humanos Par 3/genética , Cromosomas Humanos Par 7/genética , Codón sin Sentido/genética , Párpados/anomalías , Proteínas Nucleares , Factores de Transcripción/genética , Femenino , Genes Dominantes/genética , Heterogeneidad Genética , Genotipo , Humanos , India , Fenotipo , Síndrome , Proteína 1 Relacionada con TwistRESUMEN
During development, Bone Morphogenetic Proteins (BMPs) can induce apoptosis, cell growth or differentiation. These different effects are mediated by dimers of two types of BMP-receptors (BMPRs). To identify the responding cells during tooth development and search for possible tissue-or stage-specificities in the receptors involved, the distribution patterns of BMPR-IA, -IB and -II were investigated in the mouse molar, from bud to bell stage. At the bud stage, BMP-2 was suggested to be involved in the formation of an epithelial signaling center, the primary enamel knot (PEK), while BMP-4 would mediate the condensation of the mesenchyme. Immunostaining showed the presence of BMPR-IA and -II in the epithelium instead of BMPR-IB and -II in the mesenchyme. At the cap stage, BMPR-IB was detected in the epithelium but not BMPR-II, suggesting the existence of another type II receptor to form a functional dimer. At the late cap stage in the epithelium, BMP-4, BMPR-IA and -II were restricted to the internal part of the PEK and the stalk: two apoptotic areas. The three proteins were detected in the mesenchyme, showing a strong staining where cusps were about to form. At the late bell stage, BMP-2 or -4 may induce cell differentiation. BMPR-IB and -II were detected in odontoblasts instead of BMPR-IA and -II in ameloblasts. These results provide the first evidence of multiple type I and type II BMP-receptors, expressed in the dental epithelium and mesenchyme at different stages of development, to signal different cellular activities in a time- and tissue-specific way.
Asunto(s)
Receptores de Proteínas Morfogenéticas Óseas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Diente Molar/embriología , Diente Molar/metabolismo , Animales , Receptores de Proteínas Morfogenéticas Óseas de Tipo 1/metabolismo , Receptores de Proteínas Morfogenéticas Óseas de Tipo II/metabolismo , Femenino , Ratones , Ratones Endogámicos ICRRESUMEN
The twist zygotic gene appears to be involved in the establishment of the dorso-ventral pattern in Drosophila embryos. Homozygous twist embryos are partially dorsalized, their gastrulation is abnormal, and they fail to differentiate mesoderm. We determined the temperature-sensitive period of twist around the gastrulation time, and we isolated the gene. A 300 kb chromosomic walk allowed the detection of the 70 kb deletion that delimits the twist region in Df(2R)twiS60. Southern blot analyses of 21 EMS induced twist allele DNAs and systematic Northern blot analyses all over this 70 kb region lead to the localization of the twist gene: within about 10 kb at the left border of the deletion, 2 twist alleles show each a small deletion that uncover a transcription unit whose expression occurs about at the time of gastrulation.
Asunto(s)
Drosophila melanogaster/genética , Genes , Alelos , Animales , Deleción Cromosómica , ADN/análisis , Drosophila melanogaster/embriología , Embrión no Mamífero/anatomía & histología , Gástrula/anatomía & histología , Genes Letales , Transcripción Genética , Cigoto/metabolismoRESUMEN
The maternal gene dorsal encodes a nuclear protein acting as a morphogen that determines the size and fate of regions along the dorsal-ventral axis of the Drosophila embryo. From previous genetic and biochemical studies it was hypothesized that dorsal might be responsible for the activation of the zygotic gene twist. In this report, regulatory sequences required for correct spatial and quantitative expression of twist are defined, by using phenotypic rescue and studying twist-beta-galactosidase expression. In addition, by transient cotransfection assays, we show that the dorsal protein specifically activates expression from the twist promoter. We demonstrate that dorsal is a sequence-specific DNA-binding protein that recognizes a motif similar to that recognized by the mammalian transcriptional activator NF-kappa B.
Asunto(s)
Proteínas de Drosophila , Drosophila melanogaster/genética , Regulación de la Expresión Génica , Proteínas Nucleares/fisiología , Fosfoproteínas , Factores de Transcripción , Animales , Secuencia de Bases , Análisis Mutacional de ADN , Elementos Transponibles de ADN , Proteínas de Unión al ADN/fisiología , Datos de Secuencia Molecular , Morfogénesis , Regiones Promotoras Genéticas , Proteínas Recombinantes/metabolismo , Secuencias Reguladoras de Ácidos Nucleicos , Transcripción Genética , Proteína 1 Relacionada con TwistRESUMEN
The twist gene was characterized in Drosophila as being necessary at gastrulation for the establishment of the mesodermal germ layer. It codes for a nuclear DNA-binding protein that is probably a transcription factor. We have cloned and sequenced the M-twist gene of Mus musculus. The deduced proteins encoded by the Mus, Xenopus, and Drosophila twist cDNAs, respectively, show a high degree of similarity. Northern blot analyses and in situ hybridizations reveal that the 1.7-kb murine M-twist m-RNA is present at early stages, starting at 8 days post coitum, and is expressed the most at 9.5 days in the cephalic and branchial mesectoderm, in some derivatives of the mesodermal layer (sclerotoma and somatopleura), and in the limb buds.
Asunto(s)
Proteínas de Unión al ADN/genética , Ratones/embriología , Ratones/genética , Factores Reguladores Miogénicos , Proteínas Nucleares/genética , Animales , Secuencia de Bases , Clonación Molecular , ADN/genética , Drosophila melanogaster/genética , Expresión Génica , Hibridación de Ácido Nucleico , ARN Mensajero/genética , Transcripción Genética , Proteína 1 Relacionada con Twist , Xenopus laevis/genéticaRESUMEN
The twist gene is involved in the establishment of germ layers in Drosophila embryos: twist homozygous mutant embryos fail to form the ventral furrow at gastrulation and lack mesoderm and all internal organs. We have determined the sequence of the twist gene, that contains 'CAX' repeats in its 5' moiety, and codes for a protein of 490 amino acids. We have raised anti-twist antibodies that were used to study the distribution of the twist protein in whole mounts and tissue sections of wild-type embryos. Twist protein appears to be a nuclear protein at all developmental stages. It is present over both poles and in the midventral region (endoderm and mesoderm anlagen) at cellular blastoderm stage; later in development, it is detected within the mesodermal layer until its differentiation into somatopleura and splanchnopleura in which some cells are still labelled by anti-twist antibodies.
Asunto(s)
Drosophila/genética , Genes , Proteínas Nucleares/genética , Factores de Transcripción , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Blastodermo/fisiología , Núcleo Celular/metabolismo , Drosophila/embriología , Proteínas de Drosophila , Embrión no Mamífero/fisiología , Endodermo/metabolismo , Gástrula/fisiología , Mesodermo/metabolismo , Datos de Secuencia Molecular , Proteína 1 Relacionada con TwistRESUMEN
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.
Asunto(s)
Moléculas de Adhesión Celular/biosíntesis , Moléculas de Adhesión Celular/metabolismo , Incisivo/metabolismo , Amelogénesis , Animales , Moléculas de Adhesión Celular/genética , Órgano del Esmalte/crecimiento & desarrollo , Órgano del Esmalte/metabolismo , Labio/crecimiento & desarrollo , Labio/metabolismo , Ratones , ARN Mensajero/biosíntesis , KalininaRESUMEN
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.
Asunto(s)
Moléculas de Adhesión Celular/genética , Regulación del Desarrollo de la Expresión Génica , Odontogénesis/genética , Diente/metabolismo , Ameloblastos/metabolismo , Amelogénesis/genética , Animales , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/metabolismo , Femenino , Inmunohistoquímica , Hibridación in Situ , Ratones , Embarazo , Conformación Proteica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Distribución Tisular , Diente/embriología , Diente/crecimiento & desarrollo , KalininaRESUMEN
Relationships between cell-cell/cell-matrix interactions and enamel organ histomorphogenesis were examined by immunostaining and electron microscopy. During the cap-bell transition in the mouse molar, laminin-5 (LN5) disappeared from the basement membrane (BM) associated with the inner dental epithelium (IDE), and nondividing IDE cells from the enamel knot (EK) underwent a tooth-specific segregation in as many subpopulations as cusps develop. In the incisor, the basement membrane (BM) in contact with EK cells showed strong staining for LN5 and integrin alpha 6 beta 4. LN5 seems to provide stable adhesion, while its proteolytic processing might facilitate cell segregation. In both teeth, immunostaining for antigens associated with desmosomes or adherens junctions was similar for EK cells and neighboring IDE cells. Outside the EK, IDE cell-BM interactions changed locally during the initial molar cusp delimitation and on the labial part of the incisor cervical loop. Conversely, cell-cell junctions stabilized the anterior part of the incisor during completion of morphogenesis. Time and space regulation of cell-matrix and cell-cell interactions might thus play complementary roles in allowing plasticity during tooth morphogenesis and stabilization at later stages of epithelial histogenesis.