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R Minkoff

Publications and source records attributed to R Minkoff.

26 records · Page 2Linked to original sources

Autoradiographic and cytochemical studies of phagocytic cells in selected fibre tracts of the mouse periodontium.

Proliferative and protein synthetic activities of phagocytic cells of specific fibre tracts of the periodontium of C57Bl mice were employing autoradiographic techniques; these were combined with a histochemical technique for horseradish peroxidase (HRP) as a marker for phagocytic activity. Animals were injected either with [3H]thymidine as a marker for proliferative activity, or with [3H]proline as a marker for protein synthetic activity prior to HRP injection. Blocks from the maxillae of experimental and control animals were fixed, decalcified, and sectioned at 50 micrometers. These were incubated with HRP localization media, dehydrated and flat embedded in Epon 812 wafers. The entire length of the periodontium, including adjacent tooth and bone, were selectively cut from the wafers, mounted on epoxy blocks and serially sectioned at 2 micrometers. Slides containing these sections were then dipped in NTB-3 nuclear track emulsion, and after appropriate exposure times, were developed and post-stained. Sections were examined microscopically, employing an ocular grid, and phagocytic cells within each area examined were delineated as either 'fibroblast-like' (FL cells) or 'endothelial/macrophage-like' (EML cells) according to criteria such as morphology, location, orientation and proximity to a vascular channel. They were then subclassified as labelled or unlabelled with respect to the autoradiographic markers. The thymidine labelling index obtained for non-phagocytic FL cells was 3.09%; this was more than twice that for phagocytic FL cells (1.35%). Similarly phagocytic FL cells in all regions studied incorporated less than half as much [3H]proline as did their non-phagocytic counterparts. This was determined by silver grain counts over HRP-stained and unstained cells using a matched pair system. In addition, the variation of the relative number of phagocytic FL cells in specific fibre tracts suggested a relationship to functional demand. The distribution of these cells was closely related to experimentally determined rates of protein turnover. Phagocytic FL cells have a markedly reduced proliferative rate and synthesize proline-containing proteins at a reduced rate. This may reflect protein production primarily for the purpose of cell maintenance. These findings are consistent with the presence of subpopulations of fibroblasts (or fibrocytes) developmentally or functionally modified for phagocytosis; alternatively, this could signify modulation of fibroblasts from primarily biosynthetic activities to degradative functions in response to varying microenvironmental conditions.

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Autoradiography of protein turnover in subcrestal versus supracrestal fibre tracts of the developing mouse periodontium.

Eighteen-day-old C57Bl mice were injected with 5 muCi/g body weight [3H]-proline and killed at intervals of 4 h to 7 wk later. Grain counts in three fibre tracts revealed that (1) half-lives of labelled protein in the developing periodontium were much shorter than those found previously for the mature periodontium , and (2) the half-life of labelled protein in the dento-gingival region was longer than the half-lives in the trans-septal and dento-alveolar fibre tracts (half-lives: dento-alveolar = 2.5 days, trans-septal = 3.8 days; dento-gingival = 7.9 days). Eight mice were given injections of [3H]-proline on days 11, 13, 15 and 17, which encompasses the formation of the three fibre tracts, and killed 4 h, 2.5, 5 and 8 wk after the last injection. All label incorporated during the formation of the periodontium had been lost by 5 wk post-injection, showing that a stable core fibre (i.e. one which is not metabolized) was not present.

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Regional variation of cell proliferation within the facial processes of the chick embryo: a study of the role of 'merging' during development.

Variation in rates of cell proliferation along the long axis of the maxillary process, within the lateral nasal process and in the zone of attachment between these structures was analyzed employing DNA labeling indices. Chick embryos were labeled with [3H]thymidine for 1 h and processed for histology and autoradiography. The percentage of labeled mesenchymal cells was determined in delineated areas. Analysis of labeling indices indicated that rates of cell proliferation varied within each of the facial processes. Regions where rates of proliferation were maintained at elevated levels were the boundary areas of the facial processes (e.g. the anterior tip of the maxillary process) and the zones of attachment between the facial processes (e.g. between the maxillary process and the lateral nasal process). Despite the presence of elevated rates of proliferation in selected regions within the facial processes, however, the percentage of labeled cells in all areas declined with advancing developmental age. These findings support the hypothesis, proposed by Streeter and Patten, that the 'merging' of adjacent facial primordia, such as the maxillary and lateral nasal processes, is accomplished by elevated rates of cell proliferation within the zones of attachment compared to the rates of proliferation in adjacent regions.

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Cell proliferation and cell density of mesenchyme in the maxillary process and adjacent regions during facial development in the chick embryo.

Cell proliferation, as measured by DNA labeling indices was analyzed during the early development of the maxillary process. Chick embryos were labeled with [3H]thymidine for 1 h and processed for autoradiography. The percentage of labeled mesenchymal cells was determined within delineated areas in the maxillary processes and in adjacent regions. Analysis of labeling indices in each of the areas at successive stages of development demonstrated a pattern of declining rates of cell proliferation with advancing developmental age. Cell proliferation in adjacent regions declined earlier and, in some instances, faster than it did in the maxillary process. Cell density was measured in the maxillary process and the roof of the stomodeum and was found to be higher in the maxillary process throughout the period studied. Cell density and cell proliferation data were analyzed with reference to the operation of 'density-dependent inhibition' of growth as a regulatory mechanism for the observed changes. 'Density-dependent inhibition' of growth was not a satisfactory explanation for the observed differences between the maxillary process and adjacent regions.

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Cell proliferation during formation of the embryonic facial primordia.

Cell proliferation of mesenchyme in the developing primary palate of the chick embryo was analyzed by tritiated thymidine autoradiography. Pulse labeling, repeated labeling, and label dilution techniques were employed to determine generation times, transit times, growth fractions, and other parameters of the cell cycle. In vivo and in vitro studies were performed to evaluate the role of tissue interactions during outgrowth of the facial primordia. These studies indicated that initially, during early stages of primary palate formation, virtually all mesenchymal cells are in the division cycle with relatively short generation times. As development proceeds, mesenchymal cell populations in the facial primordia, such as the maxillary process, retain cycle characteristics comparable to those of the progenitor cell populations. In regions adjacent to the facial primordia, such as the roof of the stomodeum, cell cycle times become more heterogeneous and result in removal of cells from rapidly cycling cell populations into subpopulations that are cycling more slowly and that, in some instances, become quiescent. Regional analysis of cell proliferation in the maxillary process indicated that growth rates of mesenchyme differ based on proximity to the overlying epithelium. Correlative in vitro studies of epithelial-mesenchymal separation and recombination experiments in organ culture revealed that the viability of mesenchyme was dependent on the presence of epithelium and that this effect was strongly stage-dependent. These and other results lead us to the conclusion that epithelial-mesenchymal interaction is significant to the maintenance of growth rates in the facial primordia and that the effects observed are mediated, at least in part, by developmental signals at the epithelial-mesenchymal interface.

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