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Biomedical subjects

M Edidin

Publications and source records attributed to M Edidin.

At least 91 records · Page 5Linked to original sources

Maternal immunostimulation of a teratocarcinoma-derived cell line, TerCs.

Since murine teratocarcinomas and early embryos are known to share cell surface antigens, we investigated the possibility of maternal immune responses to normal pregnancy using teratocarcinoma-derived cell lines as targets. We found that an adherent cell population from both the spleen and peritoneum of syngeneically mated 129/SvSl pregnant females stimulated the uptake of [125I]iododeoxyuridine ( [125I]IUdR) by a teratocarcinoma-derived cell line, TerCs in vitro. Adherent cells from multiparous females did not stimulate the growth of other tumor cell lines. However, levels of natural anti-tumor activity detected in peritoneal cell populations of 129/SvSl virgin females were greatly reduced during pregnancy. Peritoneal cells from multiparous females with growth-stimulating activity were retained on nylon-wool columns and not eliminated by treatment with anti-theta antiserum and complement. Peritoneal cells from virgin females, treated with anti-theta antiserum and complement to eliminate cytotoxic lymphocytes, gained the ability to stimulate the uptake of [125I]IUdR by TerCs cells. [125I]IUdR uptake by cultured normal mouse blastocysts was significantly enhanced by peritoneal cells from multiparous females, while cells from age-matched virgin females had no effect. These results suggest that changes in immunocyte populations occur during pregnancy in the mouse; these changes could promote the growth of the embryo in utero.

Animals↗

Effect of microvilli on lateral diffusion measurements made by the fluorescence photobleaching recovery technique.

To consider the effect of surface microvilli on measurements of lateral diffusion by fluorescence photobleaching recovery, we have measured the diffusion of the lipid probe 3,3'-dihexadecyl indocarbocyanine iodide on the villated main body and unvillated budding polar body of unfertilized mouse eggs. On the main body we found D = (6.41 +/- 0.62) x 10(-9) cm(2)/s with (77.0 +/- 2.1)% recovery, and on the budding polar body we found D = (7.05 +/- 0.75) x 10(-9) cm(2)/s with (84.7 +/- 1.3)% recovery. We thus find only slight differences in diffusion in the two regions.

Animals↗

Diffusion rates of cell surface antigens of mouse-human heterokaryons. III. Regulation of lateral diffusion rates by calcium ions.

In mouse-human heterokaryons, the lateral diffusion of major histocompatibility (MHC) antigens in the plasma membrane is enhanced by treatment of parent cells with ouabain. Ouabain treatment is ineffective if the medium lacks calcium ion, or if Verapamil, a blocker of calcium channels, is present. The divalent ionophore A23187 also enhances lateral diffusion of MHC antigens, to the same extent as ouabain, A23187 is effective only if calcium is present in the medium. Thus it appears that increased levels of cell calcium release constraints to lateral diffusion of MHC antigens.

Animals↗

Lateral diffusion of H-2 antigens on mouse fibroblasts.

We have used fluorescence photobleaching and recovery (FPR) to measure the lateral diffusion of mouse H-2 antigens, labeled with fluorescent Fab fragments, in the membrane of cl 1d fibroblasts. Diffusion coefficients, D, vary more than 20-fold from cell to cell, though they vary no more than twofold when measured at different points on a single cell. The fraction of H-2 antigens mobile, R, also varies from cell to cell, and no lateral diffusion of H-2 antigens can be detected in approximately 20% of the cells examined. Treatment of cells with NaCN + NaF, reducing their levels of ATP reduces the proportion of cells in which no lateral diffusion can be detected. The maximum values of D seen in poisoned cells are less than those in controls. Treatment of cells with the divalent inophore, A23187, greatly increases the proportion of cells in which diffusion of H-2 is rapid, D greater than 2 x 10(-9) cm2 s-1. The data obtained on diffusion by FPR can be replotted in the form of an experiment in which lateral diffusion of H-2 antigens is measured in a population of heterokaryons. There is good agreement between this transformation and actual data on heterokaryons. Thus the two methods appear to measure the same transport process.

Animals↗

H-2 expression on a teratocarcinoma-derived cell line, TerC.

The murine teratocarcinoma-derived cell line TerC, previously thought to lack products of the major histocompatibility locus H-2, was shown to express low levels of K- and D-end H-2 specificities with the use of a sensitive cytotoxic assay. The assay, based on the ability of antibody and complement to inhibit uptake of the thymidine analogue [125I]5-iodo-2'-deoxyuridine, detected appropriate public and private specificities, as demonstrated with the use of oligospecific antisera and by absorption analyses. A series of clone of TerC varied only slightly in H-2 expression, and there was no tendency for expression to increase with time in culture; thus the low levels of H-2 were not the result of a differentiating subpopulation.

Animals↗

Lipid composition and lateral diffusion in plasma membranes of teratocarcinoma-derived cell lines.

We measured lipid lateral diffusion rates for a series of teratocarcinoma-derived and embryo-derived cell lines, using the technique of fluorescence photobleaching recovery with a fluorescent lipid probe, C16dil. The probe diffuses more rapidly in plasma membranes of embryonal carcinoma cells than in plasma membranes of teratocarcinoma-derived endodermal cell lines. When embryonal carcinoma cells are induced to differentiate by treatment with retinoic acid, diffusion constants of C16dil are reduced to levels typical of endoderm. These changes are paralleled by differences in membrane cholesterol content; membrane free cholesterol levels in embryonal carcinoma lines are approximately half those found in endodermal lines, and are markedly increased upon retinoic-acid-induced differentiation.

Animals↗

Influence of the mouse major histocompatibility complex, H-2, on liver adenylate cyclase activity and on glucagon binding to liver cell membranes.

The major histocompatibility complex of mice, the H-2 complex, regulates the steady-state level of adenosine cyclic 3',5'-monophosphate (cAMP) in liver. This effect of H-2 may be due to an effect on hormone binding to receptors. Here we show that liver membranes from animals of different H-2 types differ in their sensitivity to glucagon stimulation of adenylate cyclase and in the affinity of their receptors for glucagon. No H-2-associated differences are seen in basal, NaF-stimulated, or GMP-PNP-stimulated adenylate cyclase.

Adenylyl Cyclases↗

Effect of bleaching light on measurements of lateral diffusion in cell membranes by the fluorescence photobleaching recovery method.

The Fc receptors for IgE on rat basophil leukemia 2H3 cells were labeled with either rhodamine- or fluorescein-conjugated IgE and then with Fab fragments of anti-IgE conjugated with the other dye. Fluorescence photobleaching recovery measurements of lateral diffusion were performed with one of the dyes before and after extensive bleaching of the other dye over an entire cell. Bleaching one dye did not affect subsequent measurements made with the other dye. We thus detect no evidence for photoinduced artifacts in fluorescence photobleaching recovery experiments.

Animals↗

Redistribution of membrane proteins in isolated mouse intestinal epithelial cells.

Single mouse intestinal epithelial cells (IEC) may be isolated by the use of a combination of methods used for the isolation of IEC from other species. Isolated cells remain viable for several hours. The membrane integral enzymes alkaline phosphatase and leucine aminopeptidase of isolated IEC are localized to the brush borders of IEC in tissue and in most newly isolated IEC. With time, both enzymes are found distributed over the entire cell surface. Redistribution appears to occur by diffusion in the plane of the membrane. It is slowed, but not blocked, if cells are maintained at 0 degrees C instead of at 37 degrees C, and it is not blocked by fixation in 0.5-3% paraformaldehyde. Drugs that alter cell membrane potential or that affect cell levels of ATP enhance the rate of redistribution of the enzymes.

Alkaline Phosphatase↗