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H C Slavkin

Publications and source records attributed to H C Slavkin.

At least 145 records · Page 8Linked to original sources
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↗
PubMed1991

Human developing enamel proteins exhibit a sex-linked dimorphism.

The amelogenin protein of developing dental enamel is generally accepted to mediate the regulation of the form and size of the hydroxyapatite crystallites during enamel biomineralization (1). A genetic disorder of enamel development (amelogenesis imperfecta) has been linked to the amelogenin gene AMEL(2-3), and loci regulating enamel thickness and tooth size have been mapped to the human sex chromosomes (4). In the human genome there are two AMEL loci with one copy of the gene on each of the sex chromosomes (AMELX and AMELY), whereas in the mouse only an AMELX locus is present (5). It is presently unknown if human AMELY is transcriptionally active. These observations prompted us to examine specimens of human developing enamel for sexual dimorphism at the protein level. We report here, for the first time, a diagnosis of differences in human enamel proteins which permits the distinction of specimens according to the sex of the individual.

Ameloblasts↗
PubMed1991

Amelogenin post-secretory processing during biomineralization in the postnatal mouse molar tooth.

The primary structures, molecular genetics and biosynthesis of the amelogenin protein of the developing tooth are established, but knowledge of their subsequent post-secretory processing and its relation to enamel biomineralization is fragmentary. Preparations of tooth matrix proteins were isolated from molars (M1) of mice from birth to 15 days and analysed by SDS-PAGE and immunochemical methods. Amelogenin proteins, isolated and partially purified by HPLC, were characterized by amino acid analysis and SDS-PAGE. At birth a 26 kDa amelogenin was present that during subsequent developmental stages generated a series of 20-25 kDa amelogenins differing in apparent size by approximately 1 kDa. Amino acid analyses showed that all these amelogenins have amino-terminal TRAP sequences; analyses for both glycosylation and phosphorylation were negative. It is suggested that these post-secretory amelogenins are generated by a sequence of specific carboxy-terminal cleavages, and that the observed post-secretory processing of amelogenin is functionally linked to the structure of the enamel matrix and the control of crystallite development.

Age Factors↗
PubMed1991

Cells from Hertwig's epithelial root sheath do not transcribe amelogenin.

Recent experimental evidence has led to the interpretation that "enamel-like" material is deposited along the forming mouse molar root surface by cells of Hertwig's epithelial root sheath (HERS cells) and that this material is integral to the developmental program for cementogenesis. The experimental strategy described in this study was to examine selected developmental stages of root formation for mouse first and second mandibular molars in order to localize the cellular sites of amelogenin gene transcripts using high resolution in situ hybridization. Amelogenin is the major structural protein of coronal enamel and is highly conserved among mammalian species at the DNA and amino acid sequence level. Within the limits of sensitivity for in situ hybridization and utilizing either cRNAs or oligodeoxynucleotide probes, we were unable to localize amelogenin transcripts within HERS cells from selected developmental stages associated with mouse molar root formation. In contrast, previous studies using antipeptide antibodies have provided immuno-histochemical localization of amelogenin domains in HERS cell-derived products. For these HERS cell-derived proteins to contain both amelogenin epitopes and yet fail to yield nucleic acid hybridization signals suggests that either gene rearrangement and/or alternative processing of messenger RNAs from the structural gene locus operate to produce immunologically related motifs sharing insufficient complementarity at the nucleotide level to permit efficient detection by hybridization. It is postulated that HERS cells synthesize proteins which contain amelogenin domains and that these proteins participate during cementogenesis. However, these enamel-related proteins are neither identical to, nor collinear with coronal canonical amelogenin transcripts.

Ameloblasts↗
PubMed1990

Isolation and partial characterization of a human amelogenin from a single fetal dentition using HPLC techniques.

A strategy based on high pressure liquid chromatography (HPLC) techniques for the isolation or a principal amelogenin molecule from a single human dentition is described. A partial sequence (33 residues) for this 24 kDa amelogenin is presented and related to earlier studies of human 5 kDa tyrosine-rich amelogenin polypeptides (TRAPs). A failure to identify amino acid residue #25 (tryptophan in other amelogenins) suggests that this 24 kDa amelogenin is the progenitor of the human TRAP-2 molecule and provides further support for the possibility of several human amelogenin gene products, generated by splice-junction selection, from the single amelogenin gene in the same individual. Alternatively, multiple amelogenins may arise by expression of both the AMELX and AMELY loci.

Amelogenin↗
PubMed1990

Epidermal growth factor regulates gene expression of both epithelial and mesenchymal cells in mouse molar tooth organs in culture.

Epidermal growth factor and cis-hydroxyproline specifically inhibited synthesis of type 1 collagen, a major gene product of the differentiated dental mesenchymal cells (odontoblasts). In tandem, synthesis of enamel proteins, specific gene products of differentiated dental epithelial cells (ameloblasts), was also inhibited. Under these culture conditions, total protein synthesis in tooth organs was not inhibited but rather increased. Inhibition curves of the gene products specific for epithelial and mesenchymal phenotypes were quite similar, indicating coordinate and intimately associated regulation of gene expression under conditions that perturb cytodifferentiation.

Animals↗
PubMed1990

Regulatory issues during early craniofacial development: a summary.

During neurulation, craniofacial structures derived from the first branchial arch are determined to become maxillary, mandibular, and tongue formations. At least four interdependent developmental processes become integrated: (1) allocation of cells into specific lineages (perhaps during gastrulation); (2) regulation of time-dependent differential regulatory and structural gene expressions; (3) positional information resulting in pattern formations; and (4) morphogenesis, histogenesis, and cytodifferentiation. This review highlights progress toward understanding when, where, and how the one-dimensional genetic information encoded within DNA (deoxyribonucleic acid) is sequentially expressed into the embryonic craniofacial complex. Specifically, how might intrinsic autocrine and/or paracrine regulatory factors control the developmental program for early first branchial arch morphogenesis, histogenesis, and cytodifferentiation. Rules learned from normal development should be useful toward advancing the diagnosis, treatment, prognosis, and prevention of congenital craniofacial malformations.

Animals↗
PubMed1990

Analysis of human enamel genes: insights into genetic disorders of enamel.

A number of inherited craniofacial diseases are known to be associated with gene mutations. Inherited genetic disorders of enamel formation called amelogenesis imperfecta (AI) affect the human population with a prevalence of 1 in 14,000 in the United States. Amelogenins, the major proteins in developing enamel matrix of mammalian teeth, have been suggested to participate in normal enamel matrix biomineralization, as well as with abnormal biomineralization such as seen in AI. The complementary DNA for mouse amelogenin gene (AMEL) has been cloned, characterized, and used as a probe to establish the chromosomal locations of AMEL for mouse and man. The human AMEL gene sequences have been located to the distal short arm p22.1----p22.3 region of the X chromosome, and the pericentromeric region of the Y chromosome. An assignment of human AMEL gene to the X chromosome p22 region together with a recent assignment of the X-linked AI disease locus to the Xp22.2 region support the association of the AMEL-X gene with AI. This also leads us to propose that a mutated AMEL-X gene produces altered amelogenin polypeptide, which is defective in its ability to participate in mineralization of enamel matrix, thus giving rise to the X-linked phenotypes of AI.

Amelogenesis Imperfecta↗
PubMed1990

Cartilage, bone and tooth induction during early embryonic mouse mandibular morphogenesis using serumless, chemically-defined medium.

Studies were designed to test the hypothesis that plasma- and serum-deprived embryonic cells and tissues in vitro are capable of producing growth regulating factors which augment cartilage, bone and tooth induction during mouse mandibular process development. Embryonic mouse first branchial arch-derived mandibular processes (E11-E12, Theiler stages 18-19) or cap stage molar tooth (M1) organs (E15-E16, Theiler stage 23) expressed morphogenesis, histogenesis and cytodifferentiation (e.g., Meckel's cartilage and mandibular bone) when cultured as explants in permissive serumless and chemically-defined BGJB medium for periods up to 31 days in vitro. Organ cultures of early mandibular process explants in serumless conditions showed DNA synthesis comparable to the time- and position-restricted patterns characteristic for control in vivo development. As a paradigm for embryonic cell expression of putative growth factors, sense and antisense oligodeoxynucleotide probes corresponding to amino acids 1070-1081 for preproEGF, and antibodies directed against amino acids 348-691 of preproEGF, were used to identify and localize mRNA transcripts and translation products. Our preliminary evidence suggests that odontogenic epithelial and ectomesenchyme cells produce EGF-like products during instructive phases of tooth development. We suggest that plasma- and serum-deprived cells and tissues in vitro produce autocrine and/or paracrine growth factors which mediate embryonic mandibular morphogenesis, histogenesis and cytodifferentiation.

Animals↗
PubMed1989

Human and mouse cementum proteins immunologically related to enamel proteins.

SDS-polyacrylamide gel electrophoresis, immunoblot and amino acid composition analyses were applied to human and mouse acellular cementum proteins immunologically related to enamelins and amelogenins. In this analysis, anti-mouse amelogenin, anti-human enamelin and synthetic peptide (e.g., -LPPHPGHPGYIC-) antibodies were shown to cross-react with tooth crown-derived enamelin with a molecular mass of 72,000 Da (72 kDa), amelogenins (26 kDa), and also to four human cementum proteins (72, 58, 50 and 26 kDa) and two mouse cementum proteins (72 and 26 kDa). Each of the antibodies recognized tooth root-derived cementum polypeptides which share one or more epitopes with tooth crown-derived enamel proteins. The molecular mass and isoelectric points for crown-derived and root-derived enamel-related proteins were similar. Analysis of human and mouse cementum proteins revealed a characteristic amino acid composition enriched in glutamyl, serine, glycine, alanine, proline, valine and leucine residues; compared to the major enamel protein amelogenin, cementum proteins were low in proline, histidine and methionine. The human and mouse putative intermediate cementum proteins appear to represent a distinct class of enamel-related proteins. Moreover, these results support the hypothesis that epithelial root sheath epithelia express several cementum proteins immunologically related to canonical enamel proteins.

Amelogenin↗
PubMed1989

Human amelogenins: sequences of "TRAP" molecules.

The extracellular protein matrix of developing enamel includes a major class of proteins, the amelogenins, which are believed to be concerned in regulating enamel biomineralization. Previous studies have shown the amelogenins of the extracellular matrix to be a complex of proline-rich hydrophobic proteins which, it is suggested, arise through posttranslational and postsecretory processing of a primary ameloblast gene product. More recently, it has been shown that the human amelogenin gene is located on both the X and Y chromosomes raising the possibility that polymorphism at the level of the gene may also contribute to the observed complexity of these enamel matrix proteins. To investigate such possible amelogenin polymorphism in developing human dental enamel, individual fractionated by size-exclusion and reversed-phase high pressure liquid chromatography (HPLC). Two tyrosine-rich amelogenin polypeptides (TRAPs) of approximately 5 kDa in size were isolated from an individual human dentition and characterized by automated gas-phase sequencing. These polypeptides were found to be of 42 (TRAP-2) and 44 (TRAP-1) amino acid residues in length; TRAP-2 lacked a carboxy-terminal -Gly-Trp sequence as has previously been described for analogous bovine TRAP molecules. However, residue #25 of the human TRAP-2 sequence was refractory to sequencing, apparently differing from the Trp-25 identified in TRAP-1. These findings suggest (1) two forms of TRAP molecules, differing only by cleavage of a carboxy-terminal dipeptide, are a general feature of human and other mammalian enamel proteins, probably being derived by postsecretory cleavage from the primary extracellular amelogenin; and (2) in human developing enamel four forms of TRAPs may arise either from polymorphism at the level of the gene, or by posttranscriptional alternative splicing of amelogenin mRNAs, coupled with specific post-secretory proteolytic processing.

Amelogenin↗
PubMed1989

Localization of epidermal growth factor precursor in tooth and lung during embryonic mouse development.

The murine epidermal growth factor (EGF) precursor is a 1217 amino acid protein which contains mature EGF (amino acid residues 977-1029) as well as eight EGF-like repeats. Although the highest levels of EGF are found in the adult male mouse submandibular gland, the results of in situ hybridization studies and mRNA analyses suggest that EGF precursor mRNA is synthesized in several adult mouse tissues including the lung and the incisor. To determine if EGF precursor gene expression is intrinsic to the developmental program for either embryonic tooth or lung organogenesis, sense and antisense oligodeoxyribonucleotide probes corresponding to amino acids 1070-1081 of the precursor were used to localize cellular sites of synthesis of EGF precursor mRNA by in situ hybridization. Antibodies directed against amino acid residues 348-691 of the precursor were used in immunodetection techniques to identify either EGF precursor protein or processed derivatives. In contrast to earlier reports indicating that embryonic mouse tissues do not synthesize EGF precursor mRNA, we found that EGF precursor mRNA is present in clusters of ectoderm-, mesoderm-, and ectomesenchyme-derived cells associated with embryonic teeth and lung organs. Moreover, epitopes common to the EGF precursor were immunolocalized in both the epithelial and mesenchymal tissues of embryonic mouse tooth and lung organs. These results suggest that the EGF precursor and/or motifs contained within the precursor molecule, including mature EGF, may play an instructive or permissive role in epithelial-mesenchymal interactions pursuant to organogenesis.

Aging↗
PubMed1989

Human and mouse amelogenin gene loci are on the sex chromosomes.

Enamel is the outermost covering of teeth and is the hardest tissue in the vertebrate body. The enamel matrix is composed of enamelin and amelogenin classes of protein. We have determined the chromosomal locations for the human and mouse amelogenin (AMEL) loci using Southern blot analyses of DNA from human, mouse, or somatic cell hybrids by hybridization to a characterized mouse amelogenin cDNA. We have determined that human AMEL sequences are located on the distal short arm of the X chromosome in the p22.1----p22.3 region and near the centromere on the Y chromosome, possibly at the proximal long arm (Yq11) region. These chromosomal assignments are consistent with the hypothesis that perturbation of the amelogenin gene is involved in X-linked types of amelogenesis imperfecta, as well as with the Y-chromosomal locations for genes that participate in regulating tooth size and shape. Unlike the locus in humans, the mouse AMEL locus appears to be assigned solely to the X chromosome. Finally, together with the data on other X and Y chromosome sequences, these data for AMEL mapping support the notion of a pericentric inversion occurring in the human Y chromosome during primate evolution.

Amelogenin↗
PubMed1989

Amelogenesis in vitro: a model for studies of epithelial postsecretory processing during tissue-specific extracellular matrix biomineralization.

The extracellular matrix (ECM) of developing mammalian enamel comprises a complex of unusual epithelial-derived proteins, which appear to function in concert to initiate and propagate tissue-specific biomineralization. Following enamel protein synthesis by ameloblast cells within the enamel organ, the subsequent steps of posttranslational modification, secretion, postsecretory processing and eventual removal of these proteins from forming enamel are largely unknown. To address this issue we have designed studies to investigate the hypothesis that enamel proteins are removed from enamel and translocated into the vasculature as relatively high-molecular-weight components. We examined enamel proteins recovered from serumless medium during prolonged organ culture of mouse capstage mandibular first molars. By 21 days in vitro the tooth crown formed and dentine and enamel biomineralization were apparent. At 31 days, explants retained metabolic activity and the enamel matrix showed extensive transformation. Immunologically identified enamel proteins of 26-18 k Da were produced by cultured tooth organs, translocated from tooth explants to the culture medium, recovered from the medium and then compared to control enamel protein from in vivo preparations. Comparable postsecretory processing of the 26-k Da amelogenin protein was observed in vitro and in vivo. We speculate that the pathway reported in the present studies is comparable to the processing of the enamel protein polypeptides of the maturing enamel which occurs in vivo. The in vitro organ culture model described in this report provides an approach with which to investigate the molecular events associated with epithelial-derived postsecretory processing of ECM molecules associated with tissue-specific biomineralization.

Ameloblasts↗
PubMed1989

Lamellar body formation precedes pulmonary surfactant apoprotein expression during embryonic mouse lung development in vivo and in vitro.

The purpose of this investigation was to determine whether lamellar inclusion body (LB) formation and surfactant apoprotein (SP-35) production are directly coordinated by temporal and positional information during development. In the present study we report a comparison between embryonic B10.A mouse lung morphogenesis and cytodifferentiation in vivo with that observed during organ culture in serumless medium. Precursor LB were first detected at embryonic day 12 (E12d), and progressively larger numbers and forms were produced during subsequent differentiation of respiratory alveolar duct epithelium. SP-35 was first detected during the canalicular period (E16.5d). Lung cultures (E12d) showed pseudoglandular and canalicular periods of morphogenesis, and both ciliated epithelial and type II cell differentiation. Nonciliated cells produced increasing numbers of lamellar inclusion bodies throughout the culture period. SP-35 was detected at 9 days in vitro (d.i.v.). These observations indicate (i) precursor LB formation precedes SP-35 expression and is not dependent on apoprotein synthesis; (ii) E12d lung development in vitro using serumless medium proceeds at a rate equivalent to 0.5 days in vivo through 11 d.i.v.; and (iii) morphogenesis and differentiation occur in the absence of exogenous hormones and growth factors. The cell-cell interactions that play a role in morphogenesis and cell differentiation appear to be intrinsic to the developmental program for embryonic lung development and are likely to be mediated by autocrine and/or paracrine factors.

Apoproteins↗