Size Dependence of Ghosts from Stored Erythrocytes on Calcium and Adenosine‐Triphosphate

Abstract

Erythrocyte ATP depletion during storage of ACD blood is associated with disc-sphere transformation and a decrease in membrane viscoelastic properties. A heterogeneity within the stored cells is indicated by the presence of both biconcave and spherical cells and by bimodal derivative curves of osmotic fragility.The heterogeneity in size of the ghosts, prepared from red cells at various times of storage from ACD collected blood and its dependence on erythrocyte ATP and calcium content was studied by multichannel particle size analysis. Reconstituted ghosts were prepared by osmotic hemolysis followed by restoration of isotonicity. A single population of ghosts was obtained when fresh red cells were used. ATP depletion during blood storage was accompanied by a bimodal size distribution of erythrocyte ghosts due to the appearance of a second ghost population of smaller size. Such bimodality was not observed in the size distribution of intact stored erythrocytes. The population of smaller ghosts increased in number during prolonged storage while the normal population proportionally decreased. After 7 t o 8 weeks storage of red cells, about 50% of resultant ghosts were represented by a second population. A similar population was produced from fresh cells when CaCl, (1 x 1 0 -6~) was introduced into ghosts during hemolysis.The introduction of nucleotides (ATP, ITP, GTP, ADP, AMP) or EDTA (14 x 1 0 -s~) into ghosts during hemolysis of stored cells prevented the accumulation of small ghosts. The inhibitory effect of nucleotides was greater than predicted from stability constants for C a t + indicating their specific interaction with the membrane.Determination of the calcium content in extracts of dry ashed red cells by atomic absorption spectrophotometry revealed an increase during prolonged blood storage from 11.3f6.3 ,ug atoms/l cells (first two weeks of storage) to 22.0h7.0 (7-9 weeks of storage). The calcium content of both fresh and stored cells was quantitatively recovered from the ghosts.Kinetic data revealed that upon restoration of isotonicity, ghosts became smaller in volume than the corresponding erythrocytes but subsequently swelled to a final stable volume. The second ghost population underwent markedly little reswelling. This is indicative of increased membrane rigidity which prevents changes in volume, and decreased passive permeability to NaCl.

Energized endocytosis in human erythrocyte ghosts.

“…6) confirm the previously described Ca-linked constriction of erythrocyte ghosts (28)(29)(30). In the absence of added Mg, ghosts containing Ca and ATP remained in a sharply constricted form for at least 15 min of incubation at 370C (Fig.…”

“…Interpretation of the onset and extent of endocytosis was made independently on duplicate coded samples. In the absence of added Mg, endocytosis did not occur (7,9) and addition of Ca and ATP without Mg produced constricted ghosts (28)(29)(30) (Fig. 6A) sealed ghosts (Table IV) with a Ca/P ratio in this experiment of 0.95 (Table IV).…”

Time-course investigation of SAGM-stored leukocyte-filtered red bood cell concentrates: from metabolism to proteomics

“…35 The consistency of the role of Ca 2+ in the frame of RBC storage is strengthened by the consideration about the role of this ion in modulating the Ca-dependent K channel 36 and the influence on RBC membrane shape. 37,38 Most of the proteins detected through our 2-DE approach, both in the presence and absence of NEM, had already been reported to accumulate in RBC-leaked microand nano-vesicles, for example stomatin, ankyrin, biliverdin reductase, and 14-3-3 zeta/delta (Online Supplementary Table S1 in comparison to the findings of Bosman et al 15 ), just to mention few. The list included fragments of structural proteins, among which the cytosolic domain of band 3 deserves a special mention since it is known to represent a "respiratory metabolon" docking site at the RBC membrane, 39 and is also found in RBC vesicles.…”

Trilinolein improves erythrocyte deformability during cardiopulmonary bypass.

“…The normal shape and deformability of RBCs are maintained by intracellular ATP, low content of calcium ion and cell membrane fluidity [10][11][12][13][14]. Depletion of intracellular ATP, accumulation of calcium ion or changes in membrane fluidity is associated with a decrease in RBC deformability.…”