RBC Structure and Function

 Production of RBCs:
 (1) Fetal Life – Liver, Spleen, Bone Marrow (BM)
 (2) Child/Adult – BM. (Child-lots of the skeleton
is involved, Adult-mostly the axial skeleton is
involved).
 (3) Extreme hematologic Stress – Liver and
Spleen can revert to making RBC
("Extramedullary Hematopoiesis"). Ex: kid with
severe anemia due to thalassemia with
hepatosplenomegaly.
BM Environment: cellular proliferation and maturation.
A fine reticular meshwork supports cellular elements
as vascular sinuses course through the marrow cavity
allowing for the inflow of plasma nutrients but
retaining developing cells until they are mature.
RBCs mature around a central macrophage.
RBC Development: Stem cell Þ multipotent stem cell Þ
BFU-E Þ CFU-E Þ erythroblast Þ Þ RBC
RBC Lifespan: From earliest recognizable erythroblast to
a mature RBC it takes 3-4 days, reticulocyte (nucleus
has been extruded, but some RNA is left over) for
about 1 day, and mature RBC lives 120 days.
 Hemoglobin synthesis: Three components of hemoglobin:
(1) globin, (2) protoporphyrin, (3) iron (protoporphyrin and
iron combine to form Heme).
 Iron enters the developing RBC and ultimately enters the
mitochondria to support heme synthesis.
 In the mitochondria the first step of heme synthesis takes
place as glycine and succinyl CoA combine to form delta
aminolevulinic acid.
 Synthesis shifts into the cytoplasm but ultimately returns
to the mitochondria for final steps in the formation of
protoporphyrin IX and, eventually heme, as
protoporphyrin and iron combine.
 "Sideroblastic anemia" = Congenital absence of enzymes
along the path of proto-porphyrin synthesis may lead to
severe impairment of heme synthesis
Globin chain synthesis: various Hgb's:

 Hgb A = 2 alpha chains and 2 beta chains

 Hgb A2 = 2 alpha chains and 2 delta chains
 Hgb F = 2 alpha chains and 2 gamma chains
 Changes of Hgb throughout life: Birth – most Hgb present
is of the fetal variety
 4-6 months – gradual decline in synthesis of fetal Hgb with
a corresponding increase in Hgb A
 6-8 months – approximately 97% of Hgb is Hgb A, 2% is
Hgb A-2 and 1% is Hgb F Þ
 Clinical correlate: beta chain Hgb disorders such as sickle
cell disease do not clinically manifest until 4-6 months of
age since fetal Hgb predominates during early infancy
 Mature RBC = biconcave disk, no nucleus, can’t reproduce,
can’t produce energy (no mitoch), very little cytoplasm,
diameter = 8 m m, width = 2 m m, volume = 90 femtoliters
 Three Constituents of RBCs: RBC membrane +
internal metabolic apparatus (ie a bit of
cytoplasm) + hemoglobin
 RBC membrane: Has lipid bilayer membrane
with proteins in it. Underneath it is a
cytoskeleton of proteins allowing rubbery
elasticity with main protein being Spectrin.
Ankyrin anchors cytoskeleton to membrane.
Na/K ATPase channel is abundant on membrane
(ATP from pentose phosphate shunt – that’s why
G6PD deficient people have problems).
Membrane antigen structure – there are over
300 RBC membrane Ags. They are
Polysaccharides. (A, B, Rh, Duffy, etc).
 Embden-Meyerhof Pathway - glycolysis
from glucose to lactate, net 2 ATPs
produced and used to support membrane
ion pumps. When deficiencies of the E-M
path exist, RBC survival is reduced, leading
to hemolysis
 Methemoglobin Reductase Pathway - prevents iron
of Hgb from being oxidized, makes NADH which is
reducing power. Hgb iron must be in reduced state
(Fe+2) in order to transport O 2. The environment is
constantly generating oxidant stress and therefore a
tendency to oxidize iron to Fe3+. The methemoglobin
reductase pathway counteracts this by reducing iron
to the +2 state. Patients with methemoglobin
reductase deficiency have a substantial quantity of
methemoglobin (Hgb with iron in the oxidated state)
associated with reduced O 2 carrying capacity.
 Luebering-Rapaport Pathway - modifies affinity of
binding of Hgb and O2, makes 2,3-DPG which shifts
saturation curve to the right. This pathway is an off-
shute of the E-M pathway leading to generation of 2-3
DPG. 2-3 DPG is an important regulator of Hgb-O2
release (increased 2-3 DPG giving rise to increased O2
release). An increased rate of glycolysis leads to an
increase in intracellular 2-3 DPG concentration. When
venous blood is increasingly deoxygenated, the rate
of glycolysis increases leading to increased 2-3 DPG
production and increased O2 release to the tissues.
This is an appropriate response to ensure adequate O2
delivery.
 Hexose-Monophosphate Shunt
(Phosphoglucoate Pathway) - This pathway
couples oxidative metabolism with NADP and
glutathione reductase to provide anti-oxidant
substrate which ultimately combats the effects
of oxygen stresses (environmental,
medications). If the shunt is defective (as is the
case in patients with G6PD deficiency) oxidative
insults lead to oxidation of globin chains and
denaturation of Hgb leading to precipitates
(Heinz bodies) in the RBC, membrane damage
and, ultimately, cell death.
 P50 = PO2 at which Hgb is 50% saturated
 Right shift (O2 released easily) of the curve is
caused by: ß O2 affinity, ß pH, Ý 2,3-DPG, Ý
pCO2, Ý temp
 Left shift (O2 bound tightly) of the curve is
caused by: Ý O2 affinity, Ý pH, ß 2,3-DPG, ß
pCO2, ß temp
 Kidneys are sensors (Juxtatubular cells) for O2
delivery Þ Ý EPO
 Reticuloendothelial cells participate in the destruction of
senescent RBC's.
 Destruction of RBCs happens within reticuloendothelial cells –
NOT in the circulation. Globin and heme get recycled, porphyrin is
degraded to bilirubin which is conjugated by the liver and excreted
in the gut. Rate limiting step is conjugation. Indirect
(unconjugated) bilirubin is result if this doesn’t happen.
 Normally ~10% RBCs lyse while in circulation Þ Hgb gets released
into circulation and rapidly disassociates into alpha and beta
dimers which are bound by haptoglobin. The Hgb/haptoglobin
complex is transported to the liver. If haptoglobin is depleted, free
Hgb circulates and is filtered by the kidney. Free Hgb is either
reabsorbed by renal tubular cells or excreted as free Hgb in the
urine
 The role of the RBC in the transport of O2 from the lungs to the tissues
is central. A variety of integrated physiologic components contributes
to active O2 supply to tissues including pulmonary function, and
hemodynamic factors (C.O., regional blood flow, blood volume,
viscosity).
 Regulation of the Erythron: In the basal ideal state, blood enters the
tissue at a PO2 of 95 and exits at a PO2 of 40. Therefore, 25% of O2
transported by Hgb is release. O2 delivery may be altered by: (1) Hgb-
O2 affinity and (2) increase in the number of RBC's.
 Pulmonary function/hemodynamic factors: The lungs and heart
manifest a physiologic response in the face of decreased O2 carrying
capacity associated with anemia. Alterations in regional blood flow as
well as a marked increase in cardiac output can compensate for 50%
fall in O2 carrying capacity in the anemic patient. As a result, patients
with significant anemia frequently have tachycardia and an increased
cardiac ejection fraction.