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H C Slavkin

Publications and source records attributed to H C Slavkin.

At least 19 recordsLinked to original sources
PubMed1992

Alternative splicing of the mouse amelogenin primary RNA transcript contributes to amelogenin heterogeneity.

A heterogeneous population of amelogenin proteins is derived from a single copy of the mouse amelogenin gene. To investigate the one gene--multiple protein enigma, we designed a study to distinguish between alternative splicing and proteolytic cleavage models. A pulse of [35S]methionine labeling demonstrated that multiple amelogenins are synthesized concurrently, a result consistent with an alternative splicing mechanism. Using reverse transcription and polymerase chain reaction we cloned a segment from the 5' end of a mouse amelogenin mRNA and connected it to a previously isolated abbreviated cDNA clone. Four additional cDNAs derived from alternatively spliced amelogenin mRNAs have been cloned and characterized. The five transcripts encode amelogenins 180, 156, 141, 74, and 59 amino acids in length.

Alternative Splicing↗
PubMed1992

Characteristics of phosphorylated and non-phosphorylated dentine phosphoprotein.

Heterogeneity among the odontoblast-specific, highly phosphorylated acidic protein dentine phosphoprotein (DPP) obtained from different species has been reported by several investigators. In the present study, the apparent molecular-mass variations in rabbit and mouse DPP were investigated. Extracellular matrix (ECM) DPPs were isolated and characterized. Primary gene products, before post-translational phosphorylation, were analysed based upon translation products produced in a rabbit reticulocyte lysate cell-free system using a polyclonal mouse anti-DPP antibody. Nascent non-phosphorylated DPPs were also identified from intracellular protein extracts. Mouse and rabbit ECM phosphoproteins exhibited a 10 kDa difference in size. However, nascent intracellular or translation products from both species showed the same lower molecular mass (approx. 45 kDa). Furthermore, Northern-blot analysis showed a single mRNA of the same size in both species (approx. 1.6 kb) which contains information for a protein no larger than 50 kDa. Our results indicate that the difference in molecular mass (or electrophoretic behaviour) among DPPs from different species is due to post-translational modifications, in this case phosphorylation.

Amino Acid Sequence↗
PubMed1992

Distribution of type I collagen, type II collagen and PNA binding glycoconjugates during chondrogenesis of three distinct embryonic cartilages.

Previous studies of chondrogenesis have been focused on limb bud cartilage, whereas little is known about chondrogenic processes of other cartilages with different developmental fates. We hypothesize that cartilages with various developmental fates might show identical characteristics of chondrogenesis. The chondrogenic processes in the nasal septum, the mandible, and the limb bud of the mouse were examined by means of PNA-binding glycoconjugate, and types I and II collagen expression. Swiss-Webster mouse embryos of 11 days (E11) to 14 days (E14) gestation were fixed and processed for immuno- and lectin histochemistry. The blastema of mesenchymal cell aggregates stained positively with anti-type I collagen, but very weakly with anti-type II collagen in all three models at E12, whereas PNA bound to the blastema in the limb bud but not in nasal septum or mandible. Types I and II collagens coexisted in cartilages at E13. Type II collagen was predominant in E14; type I collagen was confined to the peripheral region. The synchronized transitional expression of the collagen phenotypes in all three embryonic cartilages may be systemically regulated. The presence or absence of the PNA-binding glycoconjugates may be involved in characterizing the nature of the cartilages.

Animals↗
PubMed1992

Polyoma virus-induced murine odontogenic tumors.

Neonatal mouse pups were injected subcutaneously with polyoma virus to induce odontogenic tumors. This treatment resulted in a spectrum of tumors that arose in organs dependent upon epithelial-mesenchymal interactions for their organogenesis, which included the teeth, salivary glands, thymus, and lacrimal glands. In addition, several odontogenic tumors with a histologic resemblance to ameloblastoma were identified and analyzed with respect to the presence of markers specific for various stages of ameloblast differentiation. Immunodetection analyses of the odontogenic tumors identified fibronectin and laminin, typical of basement membrane organization during early tooth organogenesis. These same tumors failed to express amelogenin, a gene whose expression is limited to differentiated ameloblasts. In contrast, a 47 kDa enamelin-like polypeptide was identified with the use of an antienamelin antibody. These data were interpreted to suggest that the polyoma virus truncated the differentiation pathway for these odontogenic tissues at an early stage of their development and retained the expression of basement membrane components and the enamelin-like polypeptides, yet excluded expression of amelogenin gene products. This observation suggests that polyoma viral transformation may dysregulate odontogenic tissue interactions and produce tumors composed of cells arrested at a specific stage in their development.

Ameloblastoma↗
PubMed1992

Distribution of glycoconjugates localized by peanut and Maclura pomifera agglutinins during mouse molar root development.

Glycolipids, glycoproteins, glycosaminoglycans and sialoglycoproteins have all been implicated in a number of developmentally significant processes related to complex interactions between cell surfaces and the extracellular matrix. The present study was designed to localize glycoconjugates recognized by peanut agglutinin (PNA) and Maclura pomifera (MPA) lectins during mouse molar root development. Postnatal ICR mice at 10, 15, 21, 28 and 42 days were used. Lower jaws were dissected, fixed in 4% paraformaldehyde, decalcified in 5% EDTA and embedded in paraffin. Serial sections were made and stained with FITC-conjugated PNA or MPA. beta-Lactose was used as an inhibitory sugar for PNA, and alpha-D-melibiose for MPA. PNA specifically stained Hertwig's epithelial root sheath (HERS), whereas MPA stained a number of tissues. The outermost layer of root dentin, forming cellular cementum, alveolar bone and HERS showed positive reactions with MPA. Glycoconjugates localized by the lectins may be functionally related to molecules which contribute to root formation and cemento-genesis.

Animals↗
PubMed1992

Incidence of cleft lips, palates rising.

About 1:700 live human births are affected by cleft lip and palate. Socio-economic factors--teenage pregnancies, pregnancies in women older than 35, increased consumption of teratogens during early months of pregnancy--have resulted in increased numbers of congenital malformations in the United States.

Adolescent↗
PubMed1992

Endogenous epidermal growth factor regulates the timing and pattern of embryonic mouse molar tooth morphogenesis.

The tooth organ provides a model for discrete patterns of morphogenesis over short periods of developmental time. Studies were designed to test the hypothesis that endogenous epidermal growth factor (EGF) functions to regulate multiple cusp molar tooth morphogenesis during embryonic mouse development. The relative levels of endogenous EGF and EGF receptor (EGFR) transcripts were determined in both enamel organ epithelia and dental ectomesenchyme by reverse transcription-polymerase chain reaction (RT-PCR) assays. EGF and EGFR were localized by immunohistochemistry; both antigenic determinants were demonstrated on the same odontogenic cells in cultured tooth explants. To examine EGF-mediated signal transduction, cap stage mouse molar tooth organs (E16) were cultured in serumless, chemically-defined medium as either (i) controls, or supplemented with (ii) tryphostin (an EGF receptor kinase inhibitor), (iii) tyrphostin plus exogenous EGF, and (iv) exogenous EGF. Antisense oligodeoxynucleotide (ODN) strategy was used to investigate the functions of endogenous EGF employing (i) non-treated control, (ii) sense ODN control, (iii) antisense ODN, (iv) exogenous EGF, (v) sense ODN with exogenous EGF, and (vi) antisense ODN with exogenous EGF. Tyrphostin inhibited DNA synthesis and produced a significant decrease in the volume of the explants. These effects were recovered by addition of exogenous EGF. Antisense ODN inhibition resulted in abnormal cusp formations, decreased DNA synthesis, total DNA, RNA and protein content, and decreased stellate reticulum and tooth explant volumes. The decreased tooth size was not uniform, the most pronounced effect was in the stellate reticulum. This pattern of changes was not seen when antisense ODN treatment was supplemented with exogenous EGF. These results suggest that during cap stage of odontogenesis endogenous EGF acts to stimulate DNA synthesis, which increases the cell number of specific phenotypes within the enamel organ epithelia, and thereby regulates molar tooth morphogenesis.

Animals↗
PubMed1992

Gene expression, signal transduction and tissue-specific biomineralization during mammalian tooth development.

Tooth development provides a paradigm for intrinsic molecular controls for cell- and extracellular matrix (ECM)-mediated biomineralization. The intent of this review is to evaluate the sequential timing and positional information prerequisite for tissue-specific biomineralization. Recent investigations suggest that 1,25-dihydroxyvitamin D3 functions to up-regulate VDR (vitamin D receptor) that in turn could induce structural gene products, including calcium-binding proteins and several ECM proteins (e.g., enamelins, amelogenins, dentine sialoglycoproteins (DSP) and dentine phosphoproteins (DPP)), resulting in dentine and enamel formation. Inhibition of regulatory gene products and/or their receptors likely results in hypoplastic and/or hypomineralized ECM as a direct consequence of down-regulated (1) transcription and/or translation of structural and regulatory genes, (2) posttranslational modifications, (3) and/or decreased calcium transport to the forming dentine and enamel matrices. Advances in serumless in vitro culture methodology; computer-assisted access to nucleic acid sequences for probes to define when, where, and how many specific regulatory and structural gene products are expressed; antisense oligodeoxynucleotides to inhibit specific translation; and microtechniques to analyze biomineralization all provide additional avenues to investigate tissue-specific biomineralization.

Animals↗
PubMed1992

Protein kinases in dentinogenesis.

Protein modifications such as phosphorylation and dephosphorylation are known to control several cell functions including regulation of the cell cycle, signal transduction and enzyme activation/inactivation. Bone and dentin contain highly phosphorylated anionic proteins that appear to be involved in the regulation of mineralization. This study was designed to identify and characterize the enzyme(s) responsible for phosphorylation (kinases) of dentin phosphoprotein (DPP) during dentinogenesis. DPP-protein kinase activity was demonstrated in a crude homogenate of dental pulp and odontoblast cells. In parallel studies, oligonucleotides to conserved amino acid sequences present in the active site of kinases were constructed and used to screen a lambda-gt11 tooth organ cDNA library. Several cDNA clones were isolated, the size of the insert determined by PCR (polymerase chain reaction) amplification, and in situ hybridization was used to determine cellular localization during tooth organ development. Preliminary evidence provides additional molecular determinants involved with candidate kinases responsible for DPP phosphorylation and dentinogenesis.

Animals↗
PubMed1992

Regulatory factors in pulp biology: a reaction.

A number of developmental processes can be investigated in the mammalian pulp. Of particular interests are those developmental processes related to when, where and how specific phenotypes are expressed in the forming pulp tissues. This "reaction" attempts to highlight the basic processes related to immunological factors associated with accessory cells, growth factors and the determination and differentiation of odontoblasts, and dentin-specific non-collagenous proteins. The three papers associated with these developmental processes provide an exciting glimpse into the emerging advances being made in pulp biology.

Biology↗
PubMed1992

Desmin expression during early mouse tongue morphogenesis.

Occipital somites provide progenitor cells for craniofacial muscle development including the tongue musculature. Serum-derived factors are assumed to be pre-requisite for myogenesis in vitro. To test these assertions, we designed experiments to determine whether early mouse tongue development in general, and desmin localization in particular, were expressed during the development of embryonic mouse first branchial arch explants cultured in serumless, chemically-defined medium. Immunohistochemical techniques determined the chronology and positions of desmin expression during early craniofacial development. Occipital somites expressed desmin at E9 (9 days +/- 2 h post-fertilization, 18-20 somites). A discrete cell migration pathway initiating in the somites and terminating in the lateral lingual processes of the tongue primordium was defined based upon desmin expression patterns in E9-E11 embryos and computer-assisted three dimensional reconstructions. The in vitro model system was permissive for tongue morphogenesis, allowing development and fusion of the lateral lingual processes with the tuberculum impar. During culture myoblasts were not observed to fuse into myotubes with sarcomeric assembly, even though explant myoblasts produced muscle-specific protein. E10 explants cultured for 9 days demonstrated a five-fold increase in cell number that expressed desmin (P less than 0.05) when compared to the E10 starting material. We interpret these results to indicate that the tongue myogenic cell lineage was determined between E8 and E11, and that this resident population expanded within explants cultured in serumless medium by several explanations: (i) cells other than progenitor myoblasts (e.g., satellite cells) were induced to become myoblasts, and/or (ii) progenitor myoblasts within the original explants expanded by cell division in the absence of serum factors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1979

An antisera for the fluorescent labeling of mouse amelogenesis.

Embryonic mammalian enamel extracellular matrix is immunogenic. Antisera has been produced in New Zealand white rabbits using 5-day-old (post-natal) C57B1/6J mandibular and maxillary incisor and molar tooth organs as immunogens. The expression of secretory amelogenesis in mouse molar tooth organs was studied from the "cap stage" (circa 17-day fetus) to the fifth day of postnatal odontogenesis using indirect immunofluorescent microscopy. The specificity of the antisera for enamel matrix secretion was unequivocal. Secretory amelogenesis was observed in molar tooth organs as early as day-2 postnatal age. These reagents and methods provide a significant strategy in studies of epithelial-mesenchymal interactions during tooth development.

Amelogenesis↗
PubMed1979

Amelogenesis in vitro.

The processess by which diversity is acquired and maintained during embryogenesis have been repeatedly described but are as yet not understood. One fascinating example of diversity following gastrulation is epidermal organogenesis and in particular embryonic tooth morphogenesis. Tooth morphogenesis in situ and in vitro encompasses such interesting developmental problems as epithelial-mesenchymal interactions, cytodifferentiation, dentine and enamel extracellular matrix production, mineralization, issues of form and enamel maturation. In vitro methods provide access to many of these problem areas. This presentation will review selected issues related to amelogenesis in vitro and will emphasize avenues for future research.

Ameloblasts↗
PubMed1979

Speculations regarding the influence of the major histocompatibility complex (H-2) upon congenital craniofacial malformations in inbred and congenic strains of mice.

Investigations in different mammalian species have demonstrated a remarkable degree of analogy between distant mammalian species. All mammals possess a major histocompatilibity complex. The identified and postulated functions for the MHC appear to be similar among all mammalian species [14, 25]. One fascinating, but still unconfirmed possibility is that the MHC functions to regulate the immunobiology of normal pregnancy. It should be stressed at this point that much more knowledge is necessary to elucidate the mechanism of H-2-disease associations in mice and HLA-disease associations in man [25]. One explanation of the observed H-2 and HLA-disease associations is linkage disequilibrium. Data are available suggesting a tenuous HLA--cleft lip and/or palate association [34, 35] but the data are equivocal. The available data from congenic mice studies are considerably less equivocal and strongly suggest several important future approaches. Identification of the H-2a haplotype susceptibility and the H-2b resistance to steroid-induced clefting is a major advance in this complex field [36]. Available methods are sensitive, and newer, more precise methods are now in the offing. The renaissance in the field of congenital craniofacial malformations is rather exciting, especially the renewed awareness of interdisciplinary research toward ultimate prevention. Understandably, many physicians, dentists, nurses, and pharmacists, as well as allied biomedical health professionals investigating the heredity of congenital craniofacial malformations, are deeply interested in immunogenetics. In tandem, the biomedical scientific community (eg developmental biologists, molecular biologists, immunologists) wish to learn more about the complicated and often overlapping relationships between various clinical features of the diseases concerned. All are aware that our knowledge of this fascinating subject will not stand still; many are busy changing it.

Animals↗
PubMed1978

De novo induction of a gene product during heterologous epithelial--mesenchymal interactions in vitro.

Mesenchymal specification of epithelial cytodifferentiation and morphogenesis has been considered to be a general feature of various epithelial-mesenchymal interacting systems (e.g., salivary gland, mammary gland, feather, hair, and tooth morphogenesis). In contrast, we have demonstrated that a mesenchyme can be induced by a heterologous epithelium to synthesize in quantity a specific gene product(s) unorthodox to the organ from which the mesenchyme was taken. Stage 22-23 avian limb bud epithelium induced 17-day embryonic mouse tooth mesenchyme to differentiate into cartilage. Peptide analysis (cyanogen bromide cleavage after purification of extracted collagen chains) demonstrated that heterologous tissue recombinations produced type II collagen [alpha(II)](3) (i.e., cartilage-type) in addition to type I collagen [alpha(I)](2)alpha(2). Intact or reconstituted mouse molar tooth organs synthesized type I collagen and type I trimer [alpha(I)](3) collagen. Immunohistochemical criteria using anti-type II collagen antibodies identified type II collagen in cartilage-like matrix within the mesenchymal component of heterologous tissue recombinants. Cartilage has never been described during in vivo or in vitro tooth tissue differentiation or associated with the pathology of dental papilla mesenchyme. These results support the hypothesis that epithelial-mesenchymal interactions during embryonic development can selectively induce de novo synthesis of unique gene products.

Animals↗
PubMed1978

HLA phenotype frequencies in individuals with cleft lip and/or cleft palate.

The HLA types of 133 patients with cleft lip and/or cleft palate were determined. Caucasian patients with isolated cleft palate showed a possible association with HLA. Ten of 11 male patients had HLA--A2 as compared to one out of eight female patients (P less than 0.005). Caucasian males with cleft lip and cleft palate had a slight increased frequency of HLA--Aw24 (P = 0.07) and Mexican-American males with cleft lip and/or cleft palate showed an increase of antigen HLA-A28 (P = 0.07), though neither were statistically significant. Females with cleft lip and/or cleft palate from either racial group had no differences from the controls. The serum from 90 mothers of patients with cleft lip and/or cleft palate were reacted against their child's lymphocytes. Of these crossmatch tests, 12% were found to be positive. These preliminary results suggest that male patients with isolated cleft palate are worthy of further studies with respect of HLA associations.

Cleft Lip↗