scholarly journals THE ELECTROPHORETIC MOBILITY OF RABBIT ERYTHROCYTES AND GHOSTS

1939 ◽  
Vol 22 (4) ◽  
pp. 545-553 ◽  
Author(s):  
Harold A. Abramson ◽  
Robert F. Furchgott ◽  
Eric Ponder

Measurements of the electrical mobility of washed rabbit red cells and of ghosts produced by hypotonic solutions, freezing-and-thawing, chloroform, and saponin were made in the Abramson horizontal microelectrophoresis cell. These different forms of lysis, which corresponds to a variety of degrees of injury to the red cell, are unaccompanied by any change in electrical mobility. These observations are discussed from the standpoint of the possible structure of the cell membrane and the action of lysins upon it.

1974 ◽  
Vol 64 (6) ◽  
pp. 706-729 ◽  
Author(s):  
W. R. Redwood ◽  
E. Rall ◽  
W. Perl

The permeability coefficients of dog red cell membrane to tritiated water and to a series of[14C]amides have been deduced from bulk diffusion measurements through a "tissue" composed of packed red cells. Red cells were packed by centrifugation inside polyethylene tubing. The red cell column was pulsed at one end with radiolabeled solute and diffusion was allowed to proceed for several hours. The distribution of radioactivity along the red cell column was measured by sequential slicing and counting, and the diffusion coefficient was determined by a simple plotting technique, assuming a one-dimensional diffusional model. In order to derive the red cell membrane permeability coefficient from the bulk diffusion coefficient, the red cells were assumed to be packed in a regular manner approximating closely spaced parallelopipeds. The local steady-state diffusional flux was idealized as a one-dimensional intracellular pathway in parallel with a one-dimensional extracellular pathway with solute exchange occurring within the series pathway and between the pathways. The diffusion coefficients in the intracellular and extracellular pathways were estimated from bulk diffusion measurements through concentrated hemoglobin solutions and plasma, respectively; while the volume of the extracellular pathway was determined using radiolabeled sucrose. The membrane permeability coefficients were in satisfactory agreement with the data of Sha'afi, R. I., C. M. Gary-Bobo, and A. K. Solomon (1971. J. Gen. Physiol. 58:238) obtained by a rapid-reaction technique. The method is simple and particularly well suited for rapidly permeating solutes.


Blood ◽  
1989 ◽  
Vol 74 (5) ◽  
pp. 1836-1843 ◽  
Author(s):  
G Pasvol ◽  
JA Chasis ◽  
N Mohandas ◽  
DJ Anstee ◽  
MJ Tanner ◽  
...  

Abstract The effect of well-characterized monoclonal antibodies to red cell surface molecules on the invasion of human red cells by the malarial parasites Plasmodium falciparum and Plasmodium knowlesi was examined. Antibodies to glycophorin A (GP alpha) inhibit invasion for both parasite species, and this is highly correlated with the degree to which they decrease red cell membrane deformability as measured by ektacytometry. This effect on rigidity and invasion was also seen with monovalent Fab fragments. The closer the antibody binding site was to the membrane bilayer, the greater was its effect on inducing membrane rigidity and decreasing parasite invasion. Antibodies to the Wright determinant in particular were the most inhibitory. This differential effect of the various antibodies was not correlated with their binding affinities or the number of sites bound per cell. Antibodies to surface molecules other than GP alpha were without effect. A novel mechanism is described whereby monoclonal antibodies and their Fab fragments directed at determinants on the external surface of red cells might act to inhibit invasion by malarial parasites by altering membrane material properties.


Blood ◽  
1951 ◽  
Vol 6 (4) ◽  
pp. 350-356 ◽  
Author(s):  
ERIC PONDER

Abstract Human red cells treated with trypsin in such a way as to become agglutinable in the presence of incomplete antibodies are affected in certain other respects. Their volume is slightly increased, their ghosts are unusually rigid or "volume-occupying," their osmotic and mechanical fragilities are slightly increased and their electrophoretic mobility is reduced. These changes are probably due to effects on the protein components of the red cell surface ultrastructure. Similar effects are also produced by the related enzyme mexacain.


1935 ◽  
Vol 19 (1) ◽  
pp. 35-44 ◽  
Author(s):  
Eric Ponder ◽  
Douglas Marsland

By means of measurements from cinematograph films of the time taken for human red cells to lose hemoglobin while hemolyzing, it is shown that small concentrations of saponin bring about a relatively small permeability of the cell membrane to the pigment, whereas large concentrations so destroy the membrane that the theoretical time for loss of pigment through a completely permeable membrane (0.16 second) is very nearly attained. These results are in agreement with those obtained from electrical measurements, and the dependence of permeability on lysin concentration can be explained on the basis of what is known about the rate of transformation of lysin as it reacts with the cell envelope. When cells are hemolyzed by hypotonic solutions, on the other hand, the permeability of the membrane to pigment is nearly constant, irrespective of the tonicity used to bring about lysis.


Blood ◽  
1987 ◽  
Vol 69 (2) ◽  
pp. 401-407
Author(s):  
RS Schwartz ◽  
JA Olson ◽  
C Raventos-Suarez ◽  
M Yee ◽  
RH Heath ◽  
...  

The intraerythrocytic development of the malaria parasite is accompanied by distinct morphological and biochemical changes in the host cell membrane, yet little is known about development-related alterations in the transbilayer organization of membrane phospholipids in parasitized cells. This question was examined in human red cells infected with Plasmodium falciparum. Normal red cells were infected with strain FCR3 or with clonal derivatives that either produce (K+) or do not produce (K-) knobby protuberances on the infected red cells. Parasitized cells were harvested at various stages of parasite development, and the bilayer orientation of red cell membrane phospholipids was determined chemically using 2,4,6-trinitrobenzene sulphonic acid (TNBS) or enzymatically using bee venom phospholipase A2 (PLA2) and sphingomyelinase C (SMC). We found that parasite development was accompanied by distinct alterations in the red cell membrane transbilayer distribution of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS). Increases in the exoplasmic membrane leaflet exposure of PE and PS were larger in the late-stage parasitized cells than in the early-stage parasitized cells. Similar results were obtained for PE membrane distribution using either chemical (TNBS) or enzymatic (PLA2 plus SMC) methods, although changes in PS distribution were observed only with TNBS. Uninfected cohort cells derived from mixed populations of infected and uninfected cells exhibited normal patterns of membrane phospholipid organization. The observed alterations in P falciparum-infected red cell membrane phospholipid distribution, which is independent of the presence or absence of knobby protuberances, might be associated with the drastic changes in cell membrane permeability and susceptibility to early hemolysis observed in the late stages of parasite development.


Blood ◽  
1984 ◽  
Vol 64 (5) ◽  
pp. 1129-1134 ◽  
Author(s):  
Y Yawata ◽  
T Sugihara ◽  
M Mori ◽  
S Nakashima ◽  
Y Nozawa

Abstract Membrane lipid analyses and electron spin resonance (ESR) studies of membrane fluidity were carried out on the red cells of a Japanese patient with hereditary high red cell membrane phosphatidylcholine hemolytic anemia (HPCHA). Increased amounts of phosphatidylcholine (PC) and cholesterol were found in the membrane lipids of the affected patient, despite normal plasma lipids. The order parameter of cholesterol-free pure phospholipid liposomes prepared from this patient's red cells was decreased, apparently because of the increased PC. In contrast, the order parameter of the total red cell lipid liposomes (containing free cholesterol) was essentially normal. The overall fluidity of the intact red cells was determined by ESR with a spin probe, 5-SAL. Again, the order parameters were normal in the intact red cells of the patient with HPCHA. This suggests that the concomitant increase of membrane cholesterol and phosphatidylcholine serves to maintain normal membrane fluidity in the HPCHA red cells.


Blood ◽  
1987 ◽  
Vol 69 (2) ◽  
pp. 401-407 ◽  
Author(s):  
RS Schwartz ◽  
JA Olson ◽  
C Raventos-Suarez ◽  
M Yee ◽  
RH Heath ◽  
...  

Abstract The intraerythrocytic development of the malaria parasite is accompanied by distinct morphological and biochemical changes in the host cell membrane, yet little is known about development-related alterations in the transbilayer organization of membrane phospholipids in parasitized cells. This question was examined in human red cells infected with Plasmodium falciparum. Normal red cells were infected with strain FCR3 or with clonal derivatives that either produce (K+) or do not produce (K-) knobby protuberances on the infected red cells. Parasitized cells were harvested at various stages of parasite development, and the bilayer orientation of red cell membrane phospholipids was determined chemically using 2,4,6-trinitrobenzene sulphonic acid (TNBS) or enzymatically using bee venom phospholipase A2 (PLA2) and sphingomyelinase C (SMC). We found that parasite development was accompanied by distinct alterations in the red cell membrane transbilayer distribution of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS). Increases in the exoplasmic membrane leaflet exposure of PE and PS were larger in the late-stage parasitized cells than in the early-stage parasitized cells. Similar results were obtained for PE membrane distribution using either chemical (TNBS) or enzymatic (PLA2 plus SMC) methods, although changes in PS distribution were observed only with TNBS. Uninfected cohort cells derived from mixed populations of infected and uninfected cells exhibited normal patterns of membrane phospholipid organization. The observed alterations in P falciparum-infected red cell membrane phospholipid distribution, which is independent of the presence or absence of knobby protuberances, might be associated with the drastic changes in cell membrane permeability and susceptibility to early hemolysis observed in the late stages of parasite development.


1952 ◽  
Vol 29 (4) ◽  
pp. 605-619
Author(s):  
ERIC PONDER

1. A large number of kinds of ghost, each with its own characteristic properties, can be prepared from human red cells. These ghosts, which differ from each other in their appearance as seen with phase optics, in their heat fragmentation, and probably in other respects also, are the result of the procedures used to prepare them (lysis by water as opposed to lysis in dilute saline, lysis followed by ‘reversal of haemolysis’ by phosphate or veronal buffers at various pH or by saturating the haemolysate with CO2, lysis by freezing and thawing, lysis by chemical lysins, etc.). In view of this, it is meaningless to speak of the ‘red cell ghost’ and of its structure without further specification. 2. No entirely satisfactory explanation for the differences in heat fragmentation of the various kinds of ghost has been found. The best one is that fragmentation into stable fragments occurs when the ghost is discoidal and when it has a certain plasticity, the plasticity depending on the concentration of residual Hb. It is shown that heat fragmentation of red cells occurs maximally in a tonicity of about 1.5, i.e. when the Hb molecules are a certain optimal distance apart. 3. A number of other phenomena which occur optimally at certain values of tonicity, i.e. when the Hb molecules are neither too closely packed nor too widely separated, are described. Among these phenomena are mechanical fragility, the formation of typical sickles, resistance to certain lysins, and K-Na exchange. The various forms of disk-sphere transformation seem to have no tonicity dependence.


1974 ◽  
Vol 15 (3) ◽  
pp. 555-573
Author(s):  
J. ST J. WAKEFIELD ◽  
R. M. HICKS

N-methyl-N-nitrosourea (MNU), methyl methane sulphonate (MMS), ethyl methane sulphonate (EMS) and cyclophosphamide (CP) are all cytotoxic to the transitional epithelium lining the urinary bladder, and also cause haemorrhage from the subepithelial blood capillaries. Erythrocytes enter the epithelium and are phagocytosed by the epithelial cells where they are subsequently digested. Two other compounds, dibutyl nitrosamine (DBN) and N-(4-(5-nitro-2-furyl)-2-thiazolyl)-formamide (NFTF) are only mildly cytotoxic but produce bladder epithelial tumours. Where haemorrhage occurs in these tumours, the erythrocytes are phagocytosed and digested by the neoplastic epithelial cells. The ingested red cells appear to lie free in the epithelial cell cytoplasm, surrounded by only a single unit membrane. Densitometric tracings, however, show that this membrane is derived from the epithelial cells and that the red cells lie within a phagocytic vacuole. Careful inspection reveals the red cell membrane within the dense content of the vacuole, indicating that the permeability of the red cell membrane changes rapidly as soon as it is engulfed, so that haemoglobin is released into the phagocytic vacuole. The contents of the vacuole disintegrate and myelin figures and small amounts of residual dense material appear, but at no time was it possible to demonstrate acid phosphatase activity associated with the vacuole.


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