Changes in the cation composition and active K+ transport in the red cells of fetal sheep prepartum

1985 ◽  
Vol 63 (11) ◽  
pp. 1454-1459
Author(s):  
M. W. Wolowyk ◽  
J. C. Ellory

The red blood cells of lambs, genotypically low potassium type, undergo a transition from high potassium to low potassium cell type from parturition onwards. This involves gradual changes in cell ion content, sodium pump activity, and ouabain binding. In the present study we investigated the properties of fetal red blood cells from 30 days prepartum using the chronically cannulated pregnant ewe preparation. We demonstrate that intracellular sodium increases and potassium decreases from −30 days onwards. Sodium pump activity monitored either by tracer potassium influx or ouabain binding is markedly higher in the early fetal samples examined and declines fourfold during the final month in utero. Unlike the maternal low potassium cells the early fetal red cells are refractory in terms of sodium pump stimulation by anti-L, the antibody in fact consistently inhibiting the pump. Finally, we have investigated the volume sensitivity and development of the ouabain-insensitive potassium fluxes in these cells and found that both fetal and maternal cells show a marked chloride-dependent, volume-sensitive passive potassium flux. We conclude that the decrease in active sodium transport between fetal red cells and adult low potassium cells is achieved partly by a reduction in the density of sodium pumps per cell, and then later by the introduction into the circulation of cells with Lp-antigen-modified sodium pumps.

1974 ◽  
Vol 64 (5) ◽  
pp. 536-550 ◽  
Author(s):  
John R. Sachs ◽  
Philip B. Dunham ◽  
Donna L. Kropp ◽  
J. Clive Ellory ◽  
Joseph F. Hoffman

The characteristics of the interaction of Na-K pumps of high potassium (HK) and low potassium (LK) goat red blood cells with ouabain have been determined. The rate of inhibition by ouabain of the pump of HK cells is greater than the rate of inhibition of the pumps of LK cells. Treatment of LK cells with an antibody (anti-L) raised in HK sheep by injecting LK sheep red cells increases the rate of inhibition of the LK pumps by ouabain to that characteristic of HK pumps; reduction of intracellular K (Kc) in LK cells increases the rate at which ouabain inhibits their pumps and exposure of these low Kc cells to anti-L does not affect the rate of inhibition. There is considerable heterogeneity in the pumps of both HK and LK cells in the rate at which they interact with ouabain or the rate at which they pump or both. LK pumps which are sensitive to stimulation by anti-L bind ouabain less rapidly than the remainder of the LK pumps and exposure to antibody increases the rate at which ouabain binds to the sensitive pumps; the difference between the two types of pumps disappears if intracellular K is very low. The calculated number of ouabain molecules bound at 100% inhibition of the pump is about the same for HK and LK cells. Although exposure to anti-L increases the apparent number of ouabain binding sites in LK cells at normal Kc, it does not alter the apparent number of sites in LK cells when Kc has been reduced.


1974 ◽  
Vol 63 (4) ◽  
pp. 389-414 ◽  
Author(s):  
John R. Sachs ◽  
J. Clive Ellory ◽  
Donna L. Kropp ◽  
Philip B. Dunham ◽  
Joseph F. Hoffman

The kinetic characteristics of the Na:K pump in high potassium (HK) and low potassium (LK) goat red cells were investigated after altering the intracellular cation concentrations. At low concentrations of intracellular K (Kc), increasing Kc at first stimulates the active K influx in HK cells, but at higher Kc the pump is inhibited. These results suggest that in HK cells Kc acts both at a stimulatory site at the inner aspect of the pump and by competition with intracellular Na (Nac) at the Na translocation sites. In LK cells, Kc inhibits the active K influx and the sensitivity of LK cells to inhibition is much greater than the sensitivity of HK cells. Exposure of LK cells to an antibody (anti-L), raised in an HK sheep by injection of LK sheep cells, increased the active K influx at any given Kc. The effect of the antibody was greater at higher intracellular K concentrations, and in cells with very low concentrations of K the antibody had little effect on the pump rate. The failure of anti-L to stimulate the pump in low Kc LK cells was not due to failure of the antibody to bind to the cells. Anti-L combining at the outer surface of the cell reduces the affinity of the pump at the inner surface for K at the inhibitory sites. The maximal pump rate in LK cells at optimal Na and K concentrations is less than the maximal pump rate of HK cells under the same circumstances.


1971 ◽  
Vol 57 (5) ◽  
pp. 593-609 ◽  
Author(s):  
Robert B. Gunn ◽  
Daniel C. Tosteson

2,4,6-Trinitro-3-methyl-phenol (trinitrocresol, H+TNC-) was found to inhibit anion and stimulate cation movements across the membranes of both high potassium (HK) and low potassium (LK) sheep red blood cells. The concentration of TNC- required to inhibit SO4- and Cl- efflux (10-5-10-3 M) was less than that required to increase Na+ and K+ leakage (10-3-10-2 M). Both the inhibition of anion and stimulation of cation permeation were reversed if TNC- was washed from the red cells. The cation leak caused by TNC- was much greater at 0° and 37°C than at room temperature (23°C). In sheep red cells, TNC- was found to be about 20 times more effective than salicylate and about 40 times more effective than thiocyanate in increasing cation leak. TNC- also inhibited the ouabain-sensitive potassium influx.


1990 ◽  
Vol 258 (6) ◽  
pp. C1169-C1172 ◽  
Author(s):  
J. A. Halperin ◽  
C. Brugnara ◽  
T. Van Ha ◽  
D. C. Tosteson

We have recently reported that voltage-activated fluxes of Na, K, and Ca occur in human red blood cells [J.A. Halperin, C. Brugnara, M. Tosteson, T. Van Ha, and D. C. Tosteson. Am. J. Physiol. 257 (Cell Physiol. 26): C986-C996, 1989]. The cation permeability increases progressively as the membrane potential becomes more inside positive above +20 mV. In this paper we show that this effect also occurs in high-potassium (HK), but not in low-potassium (LK), sheep and dog red blood cells. This result suggests that the voltage-activated cation transport pathway is not the result of nonspecific dielectric breakdown of the lipid bilayer but, rather, relates to some membrane component, presumably a protein, that is expressed in HK human and sheep but not in LK sheep and dog red blood cells.


1996 ◽  
Vol 271 (5) ◽  
pp. R1287-R1294 ◽  
Author(s):  
P. L. Else ◽  
D. J. Windmill ◽  
V. Markus

Previous research has shown ectotherms to have markedly lower sodium pump metabolism than endotherms. Direct measurement of enzymatic activity of the sodium pump (Na(+)-K(+)-adenosinetriphosphatase) confirmed this difference. To determine the source of this difference, sodium pump density was measured with the use of [3H]ouabain binding. Ectotherms and endotherms were found to share similar sodium pump numbers. Approximate densities (in pmol/g) were 250 for skeletal muscle, 500 for liver, 900 for heart, and 8,000 for kidney and brain. Therefore, differences in sodium pump activity between endotherms and ectotherms were due to differences in turnover rates or molecular activities of sodium pumps. Molecular activities of sodium pumps (in ATP/min) of tissues from endotherms were between 6,000 and 12,000 and, for ectotherms, between 1,500 and 2,500. Exceptions were found that included the heart of Bufo marinus. In a single invertebrate species studied, Charax destructor, the sodium pumps of the heart had a low molecular activity characteristic of ectothermic tissues. These results suggest that during the evolution of endothermy there was a general increase in the molecular activity of the sodium pump.


1978 ◽  
Vol 235 (5) ◽  
pp. C245-C250 ◽  
Author(s):  
R. G. Kirk ◽  
C. Bronner ◽  
W. Barba ◽  
D. C. Tosteson

The concentrations of potassium, sodium, and iron in human and sheep red blood cells were measured with an electron probe. Cells were prepared for analysis by spraying them on pyrolytic graphite supports. The results obtainedwith this spray technique agreed well with values measured on similar cells that were prepared for analysis by freezing, sectioning, and freeze-drying. Higher Na concentrations and lower K concentrations were found to be associated with lower cell volumes in human and high-potassium sheep cells. In low-potassium sheep cells the reverse was found, lower Na and higher K concentrations were associated with lower cell volumes. However, the amounts of iron were found to remain relatively constant in all human cells.


PEDIATRICS ◽  
1955 ◽  
Vol 15 (1) ◽  
pp. 54-62
Author(s):  
Clare N. Shumway ◽  
Gerald Miller ◽  
Lawrence E. Young

Ten infants with hemolytic disease of the newborn due to ABO incompatibility were studied. In every case the investigations were undertaken because of jaundice occurring in the first 24 hours of life. The clinical, hematologic and serologic observations in the infants and the serologic findings in the maternal sera are described. Evidence is presented to show that the diagnosis of the disorder rests largely upon the demonstration of spherocytosis, increased osmotic fragility of the red cells, reticulocytosis, and hyperbilirubinemia in a newborn infant whose red blood cells are incompatible with the maternal major blood group isoantibody and against whose cells no other maternal isoantibody is demonstrable. The anti-A or anti-B in each of the maternal sera tested in this series hemolyzed A or B cells in the presence of complement. Other serologic findings in the maternal sera were less consistently demonstrated.


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