Red blood cell volume can be independently determined in vitro using sheep and human red blood cells labeled at different densities of biotin

Blood is the major oxygen store in ducks forced to dive, and underwater endurance depends on how much of this store can be used by oxygen-sensitive tissues such as the heart and brain. Arterial injection of macroaggregated albumin labelled with technetium-99 m, which is trapped and held by capillaries, showed that circulation in dives was restricted to the thoracic and head areas. However, tracing red blood cells labelled with technetium-99 m as they were injected during dives showed not only that the time required for the activity to reach equilibrium was 4–10 times longer than when labelled cells were injected into resting ducks but also that blood flow continued in the leg and visceral regions. Tracing red blood cells, labelled with technetium-99 m and mixed in the circulation before a dive, during the dive showed that labelled red blood cells were redistributed from the peripheral and visceral areas to the central cardiovascular area. Measurement of circulating red blood cell volume during and after diving showed that, on average, 75.24 ± 4.56% of the total red blood cell volume was circulated during forced submergence. Hence, in forced dives, red blood cell volume is positioned in such a manner that the heart and brain have access to the oxygen stored there, and the residual blood flow in the periphery ensures that most of the red blood cell volume is circulated.

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Red blood cell volume and distribution before and after bleed-out in the rat

American Journal of Physiology-Legacy Content ◽

10.1152/ajplegacy.1960.198.6.1177 ◽

1960 ◽

Vol 198 (6) ◽

pp. 1177-1180 ◽

Cited By ~ 6

Author(s):

Robert J. Dellenback ◽

Gerhard H. Muelheims

Keyword(s):

Body Weight ◽

Blood Cell ◽

Red Blood Cells ◽

Cell Volume ◽

Red Blood Cell ◽

Blood Cells ◽

Before And After ◽

Negligible Contribution

The distribution of red blood cells in nine normal Nembutalized rats (323.2–415.0 gm body weight) was determined by the Cr51-labeled red blood cell technique. Microliters of red blood cells per total and per gram of tissue are reported for the testes, brain, intestine, kidney, heart-lung, spleen, liver, bone, muscle and skin. Values are also listed for the same organs and tissues determined after rapid bleed-out as found by Muelheims, Dellenback and Rawson. A comparison of these values shows that the liver, heart-lung and muscle contribute approximately 80% of all red blood cells removed in the hemorrhage. The skin, bone, kidney and intestine contribute as a group the remaining 20% with a negligible contribution from the testes and brain and no contribution from the spleen.

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Vesiculation of Red Blood Cells in the Blood Bank: A Multi-Omics Approach towards Identification of Causes and Consequences

Proteomes ◽

10.3390/proteomes8020006 ◽

2020 ◽

Vol 8 (2) ◽

pp. 6

Author(s):

Joames K. Freitas Leal ◽

Edwin Lasonder ◽

Vikram Sharma ◽

Jürgen Schiller ◽

Giuseppina Fanelli ◽

...

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Blood Bank ◽

Cell Aging ◽

Lipid Organization ◽

Membrane Components

Microvesicle generation is an integral part of the aging process of red blood cells in vivo and in vitro. Extensive vesiculation impairs function and survival of red blood cells after transfusion, and microvesicles contribute to transfusion reactions. The triggers and mechanisms of microvesicle generation are largely unknown. In this study, we combined morphological, immunochemical, proteomic, lipidomic, and metabolomic analyses to obtain an integrated understanding of the mechanisms underlying microvesicle generation during the storage of red blood cell concentrates. Our data indicate that changes in membrane organization, triggered by altered protein conformation, constitute the main mechanism of vesiculation, and precede changes in lipid organization. The resulting selective accumulation of membrane components in microvesicles is accompanied by the recruitment of plasma proteins involved in inflammation and coagulation. Our data may serve as a basis for further dissection of the fundamental mechanisms of red blood cell aging and vesiculation, for identifying the cause-effect relationship between blood bank storage and transfusion complications, and for assessing the role of microvesicles in pathologies affecting red blood cells.

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THE IN VITRO METABOLISM OF ERYTHROCYTES FROM NEWBORN INFANTS

Canadian Journal of Biochemistry and Physiology ◽

10.1139/y60-090 ◽

1960 ◽

Vol 38 (1) ◽

pp. 727-738 ◽

Cited By ~ 4

Author(s):

A. Zipursky ◽

T. LaRue ◽

L. G. Israels

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Membrane Permeability ◽

Red Blood Cell ◽

Blood Cells ◽

Newborn Infants ◽

Phosphate Esters ◽

C Storage ◽

Rapid Fall

In the red blood cells of the newborn there is a rapid fall in non-hydrolyzable phosphate during in vitro incubation. This difference appears to be due to a decreased rate of synthesis of 2,3-diphosphoglyceric acid in the erythrocyte of the newborn. In addition the incorporation of P32 orthophosphate into the red blood cell is slower in the newborn than in the adult. During 4 °C storage of blood of adults and newborns there is a progressive fall in phosphate esters which is similar in both groups.The erythrocytes of the newborn contain more potassium and water than those of adults. During storage at 4 °C the cells of the newborn lose potassium more rapidly than those of the adult. This may be related to differences in membrane permeability.

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Na- and Cl-dependent glycine transport in human red blood cells and ghosts. A study of the binding of substrates to the outward-facing carrier.

The Journal of General Physiology ◽

10.1085/jgp.93.2.321 ◽

1989 ◽

Vol 93 (2) ◽

pp. 321-342 ◽

Cited By ~ 13

Author(s):

P A King ◽

R B Gunn

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Human Red Blood Cells ◽

Glycine Transport ◽

Rapid Equilibrium ◽

Cis And Trans ◽

Best Fit ◽

Na Uptake

Na- and Cl-dependent glycine transport was investigated in human red blood cells. The effects of the carrier substrates (Na, Cl, and glycine) on the glycine transport kinetics were studied with the goal of learning more about the mechanism of transport. The K1/2-gly was 100 microM and the Vmax-gly was 109 mumol/kg Hb.h. When cis Na was lowered (50 mM) the K1/2-gly increased and the Vmax-gly decreased, which was consistent with a preferred order of rapid equilibrium loading of glycine before Na. Na-dependent glycine influx as a function of Na concentration was sigmoidal, and direct measurement of glycine and Na uptake indicated a stoichiometry of 2 Na:1 glycine transported. The sigmoidal response of glycine influx to Na concentration was best fit by a model with ordered binding of Na, the first Na with a high K1/2 (greater than 250 mM), and the second Na with a low K1/2 (less than 10.3 mM). In the presence of low Cl (cis and trans 5 mM), the K1/2-gly increased and the Vmax-gly increased. The Cl dependence displayed Michaelis-Menten kinetics with a K1/2-Cl of 9.5 mM. At low Cl (5 mM Cl balanced with NO3), the glycine influx as a function of Na showed the same stoichiometry and Vmax-Na but a decreased affinity of the carrier for Na. These data suggested that Cl binds to the carrier before Na. Experiments comparing influx and efflux rates of transport using red blood cell ghosts indicated a functional asymmetry of the transporter. Under the same gradient conditions, Na- and Cl-dependent glycine transport functioned in both directions across the membrane but rates of efflux were 50% greater than rates of influx. In addition, the presence of trans substrates modified influx and efflux differently. Trans glycine largely inhibited glycine efflux in the absence or presence of trans Na; trans Na largely inhibited glycine influx and this inhibition was partially reversed when trans glycine was also present. A model for the binding of these substrates to the outward-facing carrier is presented.

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IN VITRO INTERACTIONS BETWEEN OXYGEN AND CARBON DIOXIDE TRANSPORT IN THE BLOOD OF THE SEA LAMPREY (PETROMYZON MARINUS)

Journal of Experimental Biology ◽

10.1242/jeb.173.1.25 ◽

1992 ◽

Vol 173 (1) ◽

pp. 25-41 ◽

Cited By ~ 1

Author(s):

R. A. Ferguson ◽

N. Sehdev ◽

B. Bagatto ◽

B. L. Tufts

Keyword(s):

Carbon Dioxide ◽

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Whole Blood ◽

Blood Cells ◽

Sea Lamprey ◽

Carbon Dioxide Transport ◽

Dissociation Curves

In vitro experiments were carried out to examine the interactions between oxygen and carbon dioxide transport in the blood of the sea lamprey. Oxygen dissociation curves for whole blood obtained from quiescent lampreys had Hill numbers (nH) ranging from 1.52 to 1.89. The Bohr coefficient for whole blood was -0.17 when extracellular pH (pHe) was considered, but was much greater (-0.63) when red blood cell pH (pHi) was considered. The pHi was largely dependent on haemoglobin oxygen- saturation (SO2) and the pH gradient across the red blood cell membrane was often reversed when PCO2 was increased and/or SO2 was lowered. The magnitude of the increase in pHi associated with the Haldane effect ranged from 0.169 pH units at 2.9 kPa PCO2 to 0.453 pH units at a PCO2 of 0.2 kPa. Deoxygenated red blood cells had a much greater total CO2 concentration (CCO2) than oxygenated red blood cells, but the nonbicarbonate buffer value for the red blood cells was unaffected by oxygenation. Plasma CCO2 was not significantly different under oxygenated or deoxygenated conditions. Partitioning of CO2 carriage in oxygenated and deoxygenated blood supports recent in vivo observations that red blood cell CO2 carriage can account for much of the CCO2 difference between arterial and venous blood. Together, the results also suggest that oxygen and carbon dioxide transport may not be tightly coupled in the blood of these primitive vertebrates. Finally, red cell sodium concentrations were dependent on oxygen and carbon dioxide tensions in the blood, suggesting that sodium-dependent ion transport processes may contribute to the unique strategy for gas transport in sea lamprey blood.

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In vitro analysis of volume and pH regulation in the red blood cells of a primitive air-breathing fish, the bowfin, Amia calva

Canadian Journal of Zoology ◽

10.1139/z94-038 ◽

1994 ◽

Vol 72 (2) ◽

pp. 280-286 ◽

Cited By ~ 4

Author(s):

B. L. Tufts ◽

R. C. Drever ◽

B. Bagatto ◽

B. A. Cameron

Keyword(s):

Rainbow Trout ◽

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Disulfonic Acid ◽

Osmotic Swelling ◽

Ph Range ◽

Amia Calva

In the bowfin (Amia calva), a decrease in extracellular pH in vitro was associated with an increase in the water content and chloride concentration in the red blood cells that could be inhibited by the anion-exchange blocker, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS). After a step increase in CO2 tension, the extracellular total CO2 concentration was also significantly reduced by DIDS. Finally, over most of the experimental pH range, the red blood cell pH observed in the presence of DIDS was significantly elevated compared with that of controls. Taken together, these results indicate that as in most other fishes, chloride–bicarbonate exchange is clearly present and functional in bowfin red blood cells. Moreover, within the physiological pH range, ion movements across the anion exchanger have a marked influence on both the volume and the pH of bowfin red blood cells. In sharp contrast to the rainbow trout (Oncorhynchus mykiss), catecholamines had no effect on the volume, pH, or intracellular sodium concentration of red blood cells in the bowfin. Following osmotic swelling, rainbow trout red blood cells were able to regulate their volume back to control levels within 2 h. In the bowfin, however, there was no regulation of red blood cell volume after osmotic swelling. Thus, in contrast to many other fishes examined to date, it would appear that in the bowfin, the physiological mechanisms involved in the adrenergic response and in the regulatory volume decrease after osmotic swelling may be less active or possibly even absent in the red blood cells.

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SODIUM AND POTASSIUM MICROPROBE ANALYSIS OF ISOLATED HUMAN RED BLOOD CELLS OR RED BLOOD CELL RESEALED GHOSTS: A COMPARISON OF RESULTS BETWEEN FREEZE-DRIED OR FROZEN-HYDRATED CELLS**Part of this chapter was presented at the XXVIIth International Congress of Physiological Sciences, Paris, July 1977.

Microbeam Analysis in Biology ◽

10.1016/b978-0-12-440340-6.50025-x ◽

1979 ◽

pp. 317-325 ◽

Cited By ~ 1

Author(s):

Nicole Roinel ◽

Alain Desportes

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Microprobe Analysis ◽

International Congress ◽

Human Red Blood Cells ◽

Freeze Dried ◽

Resealed Ghosts ◽

Sodium And Potassium

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Lactate interferes with ATP release from red blood cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.01238.2006 ◽

2007 ◽

Vol 292 (6) ◽

pp. H3038-H3042 ◽

Cited By ~ 13

Author(s):

Michael D. Rozier ◽

Vincent J. Zata ◽

Mary L. Ellsworth

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Oxygen Supply ◽

Atp Release ◽

Serum Lactate ◽

Atp Production ◽

Lactate Levels

Upon exposure to low Po2, the red blood cells of most species, including humans, release increased amounts of ATP that ultimately serves as a regulator of vascular tone matching oxygen supply with demand. In pathological conditions such as malaria and sepsis, a maldistribution of perfusion exists with its severity often correlated with the extent of elevation of serum lactate frequently in the absence of an alteration in pH. We hypothesized that the increased levels of lactate might impair the ability of red blood cells to appropriately respond to conditions of low Po2, thus preventing its important blood flow regulatory role. Using an in vitro system and rabbit red blood cells, we evaluated the capacity of cells incubated with lactate to release increased amounts of ATP in response to acute exposure to low Po2. We found that in the presence of lactate, the red blood cells did not release ATP. However, when sodium dichloroacetate, a drug used clinically to lower blood lactate levels, was added, ATP release was restored to levels that were not different from that of control cells (no lactate), even though intracellular levels of ATP were not. These results support the presence of a distinct flow regulatory pool of ATP within the red blood cell that can be independently regulated, and that lactate interferes with the ATP production within this pool, thereby diminishing the amount of ATP available for release on exposure to low Po2. Therefore, if lactate levels can be reduced, the vascular regulatory capacity of the red blood cell should be restored, thus enabling the appropriate matching of oxygen supply with oxygen demand.

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Red blood cells do not contribute to removal of K+ released from exhaustively working forearm muscle

Journal of Applied Physiology ◽

10.1152/jappl.1998.85.1.326 ◽

1998 ◽

Vol 85 (1) ◽

pp. 326-332 ◽

Cited By ~ 16

Author(s):

N. Maassen ◽

M. Foerster ◽

H. Mairbäurl

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Cell Volume ◽

Red Blood Cell ◽

Blood Cells ◽

Plasma Osmolality ◽

Cell Water ◽

Forearm Exercise ◽

Hb Concentration ◽

Forearm Muscle

K+ released from exercising muscle via K+ channels needs to be removed from the interstitium into the blood to maintain high muscle cell membrane potential and allow normal muscle contractility. Uptake by red blood cells has been discussed as one mechanism that would also serve to regulate red blood cell volume, which was found to be constant despite increased plasma osmolality and K+ concentration ([[Formula: see text]]). We evaluated exercise-related changes in [[Formula: see text]], pH, osmolality, mean cellular Hb concentration, cell water, and red blood cell K+ concentration during exhaustive handgrip exercise. Unidirectional86Rb+(K+) uptake by red blood cells was measured in media with elevated extracellular K+, osmolarity, and catecholamines to simulate particularly those exercise-related changes in plasma composition that are known to stimulate K+ uptake. During exercise [[Formula: see text]] increased from 4.4 ± 0.7 to 7.1 ± 0.5 mmol/l plasma water and red blood cell K+ concentration increased from 137.2 ± 6.0 to 144.6 ± 4.6 mmol/l cell water ( P ≤ 0.05), but the intracellular K+-to-mean cellular Hb concentration ratio did not change.86Rb+uptake by red blood cells was increased by ∼20% on stimulation, caused by activation of the Na+-K+pump and Na+-K+-2Cl−cotransport. Results indicate the K+ content of red blood cells did not change as cells passed the exhaustively exercising forearm muscle despite the elevated [[Formula: see text]]. The tendency for an increase in intracellular K+ concentration was due to a slight, although statistically not significant, decrease in red blood cell volume. K+ uptake, although elevated, was too small to move significant amounts of K+ into red blood cells. Our results suggest that red blood cells do not contribute to the removal of K+ released from muscle and do not regulate their volume by K+uptake during exhaustive forearm exercise.

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