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Biomedical subjects

E J Kollar

Publications and source records attributed to E J Kollar.

At least 19 recordsLinked to original sources
PubMed1992

Alteration of murine odontogenic patterning and prolongation of expression of epidermal growth factor mRNA by retinol in vitro.

Retinoids alter the patterning of murine odontogenesis in vivo and stimulate epithelial proliferation. Because odontogenesis is dependent on proliferation of mandibular epithelium, the effects of retinol on the patterning of odontogenic epithelium were studied. These experiments control for developmental stage, applied retinoid concentration and duration of exposure. Explants exposed for 24 h to 0.1 micrograms/ml of retinol exhibited enhanced odontogenesis. Day-9 mandibles exposed to retinol at 1-5 micrograms/ml had altered epithelial patterns consistent with those in previous in vivo experiments, including supernumerary epithelial buds in regions associated with supernumerary incisors in vivo. These changes were associated with a dose-dependent increase in epithelial proliferation and a prolonged expression of epidermal growth factor (EGF) mRNA. Altered expression of EGF mRNA may be responsible for the disrupted pattern of the dental lamina. This is the first report of a retinoid-induced alteration in EGF mRNA expression.

Animals↗
PubMed1992

The correlation of temporal regulation of glycosaminoglycan synthesis with morphogenetic events in mouse tooth development.

The purpose of this study was to investigate the pattern of sulphated glycosaminoglycan synthesis during morphogenesis and cytodifferentiation in mouse tooth rudiments and to compare the results with those obtained in another study for salivary gland, a branched organ. Sulphated glycosaminoglycan was labelled by incubating molar rudiments from day 15 of gestation to day 1 post partum in medium containing [35S]-sodium sulphate. The rudiments were washed, homogenized and digested in pronase and then were sequentially digested by chondroitinase ABC and chemically degraded by nitrous acid oxidation. The fractions from each of these procedures were analysed by chromatography on Sephadex G-50 columns. The analysis revealed that, during morphogenesis, levels of chondroitin sulphate increased to a peak of 91% at day 18 and levels of heparan sulphate diminished to 8% during this period. As cytodifferentiation occurred, the level of chondroitin sulphate dropped to 64% and that of heparan sulphate increased to 35%. These results are similar to those reported for rat submaxillary gland, a branching organ. It appears that this pattern of sulphated glycosaminoglycan synthesis is not a unique feature of branching morphogenesis but may be one which marks the transition between morphogenesis and cytodifferentiation in non-branching rudiments as well.

Animals↗
PubMed1991

Expression of epidermal growth factor mRNA in the developing mouse mandibular process.

Reverse transcription and cDNA amplification (polymerase chain reaction) of total RNA preparations were used to characterize the expression of EGF mRNA in the mandibular arch of day 9-17 mouse embryos. EGF mRNA was present in mandibles at day 9 and 10 but not at days 11-17. Separate RNA preparations from epithelium and mesenchyme at day 10 revealed EGF mRNA in both components.

Animals↗
PubMed1991

EGF antisense oligodeoxynucleotides block murine odontogenesis in vitro.

The initiation of odontogenesis depends on the site-specific proliferation of mandibular epithelium beginning at Day 11 in embryonic mice. We have previously reported that the local expression of epidermal growth factor mRNA in the murine mandible is developmentally regulated, expressed at Days 9 and 10 immediately prior to the initiation of tooth bud formation at Day 11. Exposure of Day 9 mandibular explants to antisense oligomers of epidermal growth factor blocks the initiation of odontogenesis. These results are the first demonstration of the involvement of epidermal growth factor in the inductive specification of a complex epithelial derivative.

Amino Acid Sequence↗
PubMed1991

Stage-related chondrogenic potential of avian mandibular ectomesenchymal cells.

We have examined the in vitro stage-related chondrogenic potential of avian mandibular ectomesenchymal cells using micromass cultures. Our results indicate that mandibular ectomesenchymal cells as early as stage 16, soon after the formation of the mandibular arches and well before the initiation of in vivo chondrogenesis, have chondrogenic potential which is expressed in micromass culture. There is an increase in the total area of the cultures occupied by cartilage when cells from increasing stages of development are used. The nodular pattern of chondrogenesis in these cultures indicates that mandibular ectomesenchymal cells are a heterogenous population from the time of mandibular arch formation. In addition, we studied the temporal expression of the genes for extracellular matrix proteins during in vitro chondrogenesis and correlated the morphological changes with the pattern of gene expression. Low levels of type II collagen mRNA are present in the cultures prior to detection of any stainable cartilage matrix and increase 5 fold just before the onset of chondrogenesis in vitro. On the other hand mRNA for cartilage proteoglycan core protein was not detected until the second day of culture when stainable cartilage matrix was present and progressively increased thereafter. Messenger RNA for type I collagen was present at the time of initiation of cultures and continuously increased during the culture period. Our experiments also indicated that embryonic epithelia can inhibit the in vitro chondrogenesis of mandibular ectomesenchymal cells and that the inhibitory effect of embryonic epithelia is independent of its age and site of origin.

Animals↗
PubMed1991

Temporal and spatial expression of genes for cartilage extracellular matrix proteins during avian mandibular arch development.

We have examined the temporal expression of genes for extracellular matrix proteins (type I collagen, type II collagen, and the cartilage specific proteoglycan core protein) during the development of the avian mandibular arch. We detected low levels of type II collagen mRNA in the mandibular arch as early as stage 15. Type II collagen mRNA remained low but increased slightly as development progressed from stage 15 to stage 25. More dramatic increases occurred after stage 25 coincident with overt chondrogenesis. In contrast, mRNA for the core protein of cartilage specific proteoglycan was not detected prior to the onset of chondrogenesis, appeared at stage 25, and increased thereafter. Type I collagen mRNA was also present as early as stage 15 and dramatically increased after stage 28/29, coincident with initiation of osteogenesis. Using in situ hybridization, we found that type II collagen mRNA became detectable in the center of the mandible around stage 24/25 coincident with the initiation of chondrogenesis. At later stages (26-32) type II collagen mRNA was localized in the cartilaginous rudiment. The pattern of hybridization observed with the proteoglycan core protein probe at later stages of development was essentially identical to that observed with the type II collagen probe. In contrast, the probe for the alpha 1 (I) collagen mRNA was localized over the perichondrium, over differentiated bone, and in areas within the mandibular arch where bone formation had been initiated.

Animals↗
PubMed1990

Interactions between cloned gingival or periodontal ligament cells and oral epithelial cells in vitro.

Interactions between epithelial cells and fibroblast clones were examined to determine whether various fibroblasts clones derived from gingival or periodontal tissues responded differently to oral epithelial cells. We provide evidence that epithelial root sheath (ERS) cells enhanced collagen type I (CI) expression in most periodontal clones, whereas ERS cells variably influenced CI expression of gingival fibroblast clones. Modulation of collagen type III (CIII) expression in both gingival and periodontal clones by ERS cells was less in magnitude and mostly suppressive in gingival clones. Fibronectin (Fn) expression in many gingival and periodontal clones was decreased in cells associated with ERS cells. On the other hand, the influence of gingival epithelial cells on fibroblast clones tended to be inhibitory, especially in periodontal clones. Thus, gingival epithelial (GE) cells suppressed the expression of collagen types I and III and Fn in most periodontal clones. Except for suppression of Fn expression, GE cells had less influence on CI and CIII expression in gingival clones. The modulations of fibroblast extracellular matrix components by ERS and GE have profound implications for the regulation of the development, repair and regeneration of the periodontal tissues.

Clone Cells↗
PubMed1989

Retrovirus-induced insertional mutation in Mov13 mice affects collagen I expression in a tissue-specific manner.

In the Mov13 mouse mutant, transcription of the alpha 1 (1) collagen gene is blocked by a retroviral insert in the first intron. We now report that teeth derived from homozygous embryos produce a dentin layer containing normal amounts of collagen 1. In situ hybridization and RNAase protection experiments indicate that the mutant allele is efficiently transcribed in odontoblasts, in contrast to other cell types. Correct splicing of the primary transcript containing the viral sequence results in a functional alpha 1 (1) collagen mRNA. The absence of a mutagenic effect in odontoblasts, as opposed to fibroblasts, suggests that the retroviral insert interferes with tissue-specific transcriptional control of the alpha 1 (1) collagen gene, most likely by inactivating cell-type-specific cis-acting regulatory elements.

Animals↗
PubMed1989

Differentiation of odontoblasts in grafted recombinants of murine epithelial root sheath and dental mesenchyme.

The epithelial root sheath (ERS) was isolated from first molars of 5-day-old post-natal CD-1 mice using trypsin. After isolation, ERS cells remained viable in vitro and immunohistochemical examination of cultures confirmed the epithelial phenotype and the absence of mesenchymal contamination. Recombinants of isolated ERS and dental papilla resulted in odontoblast differentiation within cells of the dental papilla, and the formation of root-like fragments of dentine after 2 weeks of intra-ocular grafting. These findings indicate the inductive influence of the ERS on dental papilla cells.

Animals↗
PubMed1987

The induction of odontogenesis in non-dental mesenchyme combined with early murine mandibular arch epithelium.

First and second branchial arches were dissected from mouse embryos of 9-13 days gestational age. The epithelial and mesenchymal components were separated after enzymic digestion. Scanning electron microscopy did not reveal a dental lamina along the dental arches before day 12, after which the lamina was formed locally in the incisor and molar regions. There was no epithelial down-growth in the diastema region. Heterotypic recombinations of mandibular arch and second branchial arch tissues showed that early mandibular arch epithelia, before day 12, have odontogenic potential and can elicit the formation of a dental papilla in non-odontogenic, neural-crest-derived mesenchymal cells of the second arch. However, the mandibular mesenchyme must interact with mandibular epithelium in order to have the competence to induce teeth in non-odontogenic epithelium.

Animals↗
PubMed1986

Tissue interactions in development of teeth and related ectodermal derivatives.

A number of traditional techniques have been used to examine epithelial-mesenchymal interactions; they remain to this day the only means to examine the problems of tissue interactions. Problems at the tissue level of organization require that the approaches to these problems ask questions and use techniques that address the issues of tissue organization and cellular morphology in the context of three-dimensional organization. Manipulating the embryo is exactly that--manipulating a three-dimensional organized organism that must express its differentiated functional organs and organ systems. When the parallel and correlative studies of cell biological function achieve new insights, they will have to be tested in terms of the cellular response at the tissue level of organization.

Animals↗
PubMed1979

Tooth morphogenesis: the role of the innervation during induction and pattern formation.

The hypothesis is discussed that the innervation of the early mandibular and maxillary processes influences the initiation and patterning of tooth germs. Silver staining of embryonic mouse tissue supports the notion that the innervation is present before tooth buds appear. Data from other experimental studies is discussed in support of this hypothesis. The importance of the early events of tooth morphogenesis in initiating and patterning the dentition is discussed in the context of tooth morphogenesis as a whole. The processes involved in odontogenesis are categorized as Phase I: Initiating events; Phase II: Histogenetic events; and Phase III: Cytodifferentiative events. The complications of the interrelationship of these three seemingly discrete segments of tooth development are discussed in the context of inductive tissue interaction.

Ameloblasts↗
PubMed1978

In vitro inhibition of mouse dental development by tetracycline.

Embryonic molars and incisors were dissected from mandibles of 15-day post-fertilization C57BL/10 mouse embryos and were cultured in vitro for six days on agar-solidified Eagle's basal medium. Experimental explants were cultured on medium which was the same as the control except that 50, 75 or 100 microgram/ml tetracycline was added. Treated explants of both incisors and molars were suppressed in development and reduced in size. Enamel organs and dental papillae of all tooth germs subjected to higher tetracycline concentrations were abnormal in structure and differentiation of ameloblasts and odontoblasts was inhibited. Explants treated with higher dosage levels of the drug were more severely affected than those exposed to lower concentrations. Recovery from the suppression induced by tetracycline was observed in explants transferred to control medium for four days of growth following treatment. Differentiated ameloblasts and odontoblasts observed in the recovering tooth germs indicated that the inhibition in development was temporary. The results of this study showed that tetracycline can alter dental development in vitro prior to mineralization. The observed inhibition may be related to a disruption of collagen biosynthesis which is thought to play a role in the controlling epithelial-mesenchymal interaction involved in tooth germ morphogenesis.

Animals↗
PubMed1978

Protease secretion during onset of development in Dictyostelium discoideum.

At the onset of development, the single cells of the eukaryotic micro-organism Dictyostelium discoideum secrete proteolytic activity which can be assayed using the insoluble substrate remazolbrilliant blue hide. The activity is not secreted by exponentially growing cells, but does appear extracellularly at the onset of the stationary growth phase. When growth phase cells are resuspended in non-nutrient buffer, proteolytic activity begins to appear outside the cells. It accumulates in the buffer at a rate similar to that observed for 2 glycosidases of lysosomal origin and reaches a maximum after about 2 h of incubation. After 3--4 h incubation, centrifugation of the non-nutrient buffer removes the cells, producing a supernatant which we refer to as conditioned medium. Subsequent experiments with conditioned medium showed: (a) its incubation with purified plasma membranes results in the release of polypeptides which can be recovered and, when displayed on polyacrylamide gels, can be shown to be stage specific; and (b) that conditioned medium can decrease the rate of detachment of cells from a collagen substratum. Both effects can be prevented by the addition of remazolbrilliant blue hide suggesting that they are due to proteolytic activity present in the conditioned medium. Finally, we were able to show that conditioned medium contains components which, when spread over the bottom of plastic Petri dishes, enhance the rate of multicellular structure formation. Additional studies showed that this effect of conditioned medium could also be brought about by components which remained behind on uncoated plastic dishes after the removal of a D. discoideum cell layer. These data may be accommodated to a model in which the protease secreted during the onset of development acts on the cell membrane releasing components which coat the substratum and facilitate migration and multicellular structure formation.

Cell Membrane↗