The release of soluble H-2 alloantigens during disaggregation of mouse embryo tissue by a chelating agent

Development ◽  
1966 ◽  
Vol 16 (3) ◽  
pp. 519-530
Author(s):  
Michael Edidin
Keyword(s):  
Molecular Weight ◽  
Ethylenediaminetetraacetic Acid ◽  
Divalent Cations ◽  
Weight Bearing ◽  
Chelating Agent ◽  
Cell Aggregation ◽  
Sponge Cell ◽  
Antigenic Sites ◽  
Dissociated Cells ◽  
Embryo Tissue

Treatment with chelating agents binding divalent cations has been found to effect the dissociation of a variety of tissues of both embryo and adult animals (reviewed in Steinberg, 1958). In the course of dissociation it appears that materials are released from cell surfaces which play a part in their specific adhesion, and which may be shown experimentally to promote selectively the re-aggregation of dissociated cells (Humphreys, 1963; Moscona, 1963). The extracted materials appear to be glycoprotein complexes (Humphreys, 1965), made up of fairly small subunits, estimated to be of 13000–20000 molecular weight (Margoliash et al. 1965). Units of about the same size appear to be the antigenic sites involved in the blocking of sponge cell aggregation by rabbit anti-sponge serum, specific for a given sponge species (MacLennan, 1963). I shall here present evidence that materials of similar molecular weight bearing immunological specificities of the H-2 alloantigen system are released from the tissues of certain mouse embryos during the course of their dissociation by the chelating agent Versene (ethylenediaminetetraacetic acid).

scholarly journals Surface Responses in Cultured Fibroblasts Elicited by Ethylenediaminetetraacetic Acid

1958 ◽  
Vol 4 (3) ◽  
pp. 243-250 ◽  
Author(s):  
Ernst J. Dornfeld ◽  
Alfred Owczarzak
Keyword(s):  
Ethylenediaminetetraacetic Acid ◽  
Divalent Cations ◽  
Chelating Agent ◽  
Time Lapse ◽  
Cellular Responses ◽  
Cultured Fibroblasts ◽  
Surface Activities ◽  
Dividing Cells ◽  
Chick Heart ◽  
And Migration

Cultures of chick heart fibroblasts were perfused with the chelating agent ethylenediaminetetraacetic acid (EDTA). Cellular responses were observed under phase optics and recorded by time-lapse cinemicrography. In interphasic fibroblasts, EDTA induces cellular contraction followed by continuous protrusion and retraction of ectoplasmic blebs ("surface bubbling"), formation of motile vermiform processes, and production of rotatory ectoplasmic swellings. The contraction and surface bubbling closely resemble the metaphase contraction and "anaphase bubbling" normally displayed by cultured fibroblasts. In dividing cells, EDTA does not affect metaphases, but anaphase bubbling appears and persists; telophasic expansion and migration of daughter cells are prevented. Initiation of new mitoses occurs during and after exposure to EDTA. No cellular responses are induced by calcium, magnesium, or ferrous chelates of EDTA. The EDTA elects are completely reversible on removal of the chelating agent, resulting in the restoration of the normal interphasic cell form and the normal expansion and migration of mitotic products. The EDTA effects are interpreted to result from the chelation and removal of divalent cations from the cell surface. Possible relations to surface activities observed in normal mitosis are considered, and an hypothesis is presented regarding the role of the developing spindle in cation transfer.

A ligand-receptor model for the cohesive behaviour of Dictyostelium discoideum axenic cells

1979 ◽  
Vol 37 (1) ◽  
pp. 157-167
Author(s):  
A.R. Jaffe ◽  
A.P. Swan ◽  
D.R. Garrod
Keyword(s):  
Molecular Weight ◽  
Stationary Phase ◽  
Chelating Agent ◽  
Cell Types ◽  
Low Molecular Weight ◽  
Receptor Model ◽  
Phase Growth ◽  
Developmentally Regulated ◽  
Receptor System ◽  
Contact Sites

Axenically grown cells of D. discoideum Ax-2 harvested in the log phase of growth, cohere rapidly when shaken in phosphate buffer. After 3.5 days in the stationary phase of growth, cells become completely non-cohesive. Although they do not stick to each other, stationary phase cells do stick to both log phase cells and aggregation-competent cells. The cohesion of stationary phase cells with these other 2 cell types is inhibited by both EDTA and the low-molecular-weight factor which we have previously demonstrated in stationary-phase growth medium. There is a decline in the sensitivity of slime mould cell cohesion to the low-molecular-weight inhibitory factor as the cells become aggregation-competent. This effect parallels the developmentally-regulated decline in sensitivity to EDTA. The low-molecular-weight inhibitor is not a chelating agent, however. The effect of the inhibitor seems to be specifically against contact sites-B mediated cohesion. We suggest that the simplest cohesive mechanism which can explain our results, is that the EDTA-sensitive cohesion of log phase cells could be dependent on a ligand-receptor system.

Effects of Cytochalasins on the Initial Aggregation in Vitro of Embryonic Chick Cells

1974 ◽  
Vol 14 (1) ◽  
pp. 187-196
Author(s):  
J. C. APPLETON ◽  
R. B. KEMP
Keyword(s):  
Cytochalasin B ◽  
Cell Aggregation ◽  
Cellular Injury ◽  
Cellular Motility ◽  
Carbon Dioxide Evolution ◽  
Site Of Action ◽  
Dissociated Cells ◽  
Cytochalasin A ◽  
Final Confirmation

The initial aggregation of trypsin-dissociated cells from the skeletal muscle tissue of 9-day-old chick embryos in the presence of cytochalasins A and B was studied in order to discover the effects of these agents on contact and adhesion. Cytochalasin B (3 µg/ml) had a negligible effect on the rate of aggregation of cells over an 8-h period, but cytochalasin A at concentrations between 3 and 20 µg/ml markedly inhibited aggregation. Both agents altered the shape and size of aggregates and caused cells at their periphery to appear more spherical. The oxygen uptake of the treated cells was not noticeably different from that of the controls, despite the severe inhibition of isotopic carbon dioxide evolution. The effect of cytochalasin B on cell aggregation was reversible and although the cytochalasin A effect could not be abolished on return to medium free of A, the unaltered oxygen consumption was taken as an indication that permanent cellular injury did not occur. The effect of the cytochalasins on aggregate structure was interpreted on the basis of arrested cellular motility, but the singular inhibition by cytochalasin A of the rate of aggregation must await final confirmation of its site of action.

Red Cell Aggregation Induced by a High Molecular Weight Gelatin Plasma Substitute

1971 ◽  
Vol 3 (6) ◽  
pp. 428-435 ◽  
Author(s):  
P.M. Scholz ◽  
J. Engeset ◽  
N.A. Matheson ◽  
U.F. Gruber
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Cell Aggregation ◽  
Red Cell ◽  
Plasma Substitute ◽  
Red Cell Aggregation

Extracellular alkaline phosphatase from marine bacteria: purification and properties of extracellular phosphatase from a marine Pseudomonas sp.

1980 ◽  
Vol 26 (7) ◽  
pp. 833-838 ◽  
Author(s):  
Hiromi Kobori ◽  
Nobuo Taga
Keyword(s):  
Molecular Weight ◽  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Marine Bacteria ◽  
Divalent Cations ◽  
Maximal Activity ◽  
Pseudomonas Sp ◽  
Purification And Properties ◽  
Extracellular Alkaline Phosphatase ◽  
Increased Pressure

Extracellular alkaline phosphatase produced by a marine Pseudomonas was purified to electrophoretic homogeneity. The molecular weight of the enzyme was estimated to be 100 000. The enzyme had maximal activity at pH 11.5. The enzyme was completely inhibited by 1 mM EDTA. However, divalent cations reversed the enzyme inhibition and their order of effectiveness on the reaction was Zn2+ > Ca2+ > Mn2+ > Mg2+ > Sr2+ > Co2+. The enzyme activity was affected by the species of anion whose order of effectiveness was demonstrated to follow the lyotrophic series, Cl− > Br− > NO3−> ClO4− > SCN−. The activity of phosphatase was accelerated linearly by increased pressure until up to 1000 atm (1 atm = 101.325 kPa), and the enzyme activity at 1000 atm was 3.2 times higher than that at 1 atm.

scholarly journals THE CHEMICAL NATURE OF MACROPHAGE RNA-ANTIGEN COMPLEXES AND THEIR RELEVANCE TO IMMUNE INDUCTION

1969 ◽  
Vol 130 (3) ◽  
pp. 557-574 ◽  
Author(s):  
Georges E. Roelants ◽  
Joel W. Goodman
Keyword(s):  
Amino Acids ◽  
Complex Formation ◽  
Chemical Nature ◽  
Divalent Cations ◽  
Chelating Agent ◽  
Total Absence ◽  
Synthetic Polypeptides ◽  
Antigen Complex ◽  
Anionic Groups ◽  
Do So

10 different compounds, including natural and synthetic polypeptides, proteins, polysaccharides, amino acids, and steroid hormones, were assayed for their capacity to form complexes with peritoneal exudate cell RNA. Only molecules carrying negatively charged groups were able to do so. The formation of RNA-antigen complexes was unrelated to the immuno-potency of the "antigen," was not an enzyme-dependent reaction, did not require the synthesis of RNA following introduction of the antigen, did not seem to involve antigen-specific RNAs, was not specific for macrophages, since HeLa cells could be used as effectively, and occurred when purified RNA was mixed with antigen only in the presence of divalent cations. The complexes were very stable, once formed, but could be dissociated by exhaustive dialysis against buffers containing a chelating agent. The macrophage RNA-antigen complex therefore appears to be a chelate between anionic groups on the two components. Based on the total absence of a relationship between immunogenicity and the capacity to form such complexes, as well as the nonspecific nature of complex formation at every level examined, it appears unlikely that RNA-antigen complexes play a physiologically significant role in immune induction.

scholarly journals Removal of tetracyclines in wastewater : accumulation and distribution of chlortetracycline in bulk water and biomass compartments in activated sludge

2021 ◽  
Author(s):  
Ruba F. Farkh
Keyword(s):  
Microbial Community ◽  
Activated Sludge ◽  
Ethylenediaminetetraacetic Acid ◽  
Divalent Cations ◽  
Bulk Water ◽  
Microbial Cells ◽  
Initial Concentration ◽  
Anoxic Conditions ◽  
Polymeric Substance ◽  
The Impact

A study was conducted to examine the removal of chlortetracycline and its distribution and accumulation in three compartments; bulk water, extracellular polymeric substance (EPS) and the microbial cells in activated sludge. Also the effect of different environmental conditions on the distribution and accumulation in the three compartments was investigated. Effluent samples collected from a municipal activated sludge treatment system were set up in bath experiments to test the distribution and accumulation of chlortetracycline under aerobic and anoxic conditions for 14 days. In addition, the impact of the activity of the microbial community on the amassing of the antibiotic in the biomass was examined. The effect of divalent cations on import and accumulation of chlortetracycline was tested. Sorption in believed to be the main removal pathway in wastewater treatment systems for tetracyclines in general and chlortetracycline in particular. In this study that notion was confirmed, and it was found that the removal via sorption under anoxic condition (43.2%) is almost double of that under aerobic conditions (27.0%). The amount of what accumulated in the cells compared to that sorbed in the EPS is twice as much in the former and triple as much in the latter. These findings suggest that changes in the structure and charge of the EPS could be the reason of higher accumulation in the polymeric substance. The impact of microbial activity on the sorption and distribution of the chlortetracycline in the three compartments showed almost a similar behaviour to that under aerobic and anoxic conditions. It was clear that the more viable the microbial community, the less the antibiotic accumulated in the [sic] both biomass compartments; the EPS and microbial cells. Biomass with inhibited respiration accrued 90% of the initial concentration; where as the active microbial community was more resistant and only 24.2% of the initial concentration accumulated within the cells. The findings suggest that the antibiotic makes its way to the cells thus bypassing the EPS, and is trapped in the EPS as it is pumped out of the cells in an energy dependent mechanism. The presence of ethylenediaminetetraacetic acid (EDTA) which is a strong chelator had no import effect. Nevertheless it did indicate that the accumulation in the EPS could be attributed to the presence of cations since there was a high negative correlation (-0.98) between the disappearance of the antibiotic from the EPS compartment and the EDTA concentration used in incubation.

Abolition by Myosin and Heavy Meromyosin of the Inhibitory Effect of Smooth-Muscle Actomyosin Antibodies on Cell Aggregation In Vitro

1973 ◽  
Vol 12 (2) ◽  
pp. 631-639
Author(s):  
R. B. KEMP ◽  
B. M. JONES ◽  
U. GRÖSCHEL-STEWART
Keyword(s):  
Smooth Muscle ◽  
Striated Muscle ◽  
Cell Aggregation ◽  
Inhibitory Effect ◽  
Regulatory Function ◽  
Rabbit Serum ◽  
Heavy Meromyosin ◽  
Γ Globulins ◽  
Dissociated Cells

The ability of anti-chicken smooth-muscle actomyosin γ-globulins (anti-GAM) to inhibit the aggregation of dissociated cells from the skeletal muscle and liver of chick embryos was abolished by pretreatment of the anti-GAM with either myosin or heavy meromyosin (HMM). When the same cells were treated with HMM at a concentration of 1 mg per 2 x 106 cells/ml Eagle's MEM they aggregated as readily as untreated cells. The negative electrophoretic mobility of the embryonic chick fibroblastic cells was significantly reduced by the globulin fraction of anti-GAM but not of HMM-treated anti-GAM or non-immunized rabbit serum. Anti-chicken striated muscle actomyosin γ-globulins slightly reduced negative mobility but HMM had no effect. The experiments show that the inhibitory effect on cell aggregation of anti-GAM preparations is produced by the anti-myosin antibodies. They also provide support for the theory that a surface-localized myosin-like protein has a regulatory function in cell adhesion.

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