Physiology of the Red Blood Cells

P H Y S I O L O G Y
Dr. Maria Verena R. Remudaro • April 19, 2020
A.Y. 2019-2020 • 2nd Semester (Finals)
Transcribers: Cardenas, Carig, Casem, Castellano

OUTLINE
I. RED BLOOD CELLS
A. CHARACTERISTICS
II. RBC PRODUCTION
A. GENESIS OF RBC
B. REGULATION OF RBC PRODUCTION
C. STIMULATION FOR EPO PRODUCTION
a. Function of EPO
D. VITAMIN B12 AND FOLIC ACID
III. ANEMIA
A. CAUSES OF MEGALOBLASTIC ANEMIA
IV. HEMOGLOBIN
A. HEMOGLBIN FORMATION
B. GLOBIN SYNTHESIS
C. HEMOGLOBIN MOLECULE
V. IRON

II: RBC PRODUCTION

I: RED BLOOD CELLS
 Areas of the body that produce RBCs
A. CHARACTERISTICS
 Early weeks of embryonic life – yolk sac
 Also known as erythrocytes
 Middle trimester of gestation – liver, spleen & lymph
 Most abundant cells of the blood
nodes
 Are biconcave discs
 Last month of gestation and after birth – bone marrow
 Concentration:
 Bone marrow of all bones produces RBCs until the 5 y/o
o Men – 5,200,000 (+ 300,000)
 The marrow of long bones becomes fatty & produces no
o Women – 4,700,000 (+ 300,000)
more RBC’s after age of 20, except proximal portions of
 Life span – 120 days
humeri and tibae
 Biconcave- because they lose their nucleus & other
 >20 y/o, most RBCs are produced in the marrow of the
organelles, makes them more pliable, thus prevent
membranous bones such as vertebrae, sternum, ribs,
rupture
and ilia
 Functions:
 Bone marrow becomes less productive as age
o Transport hemoglobin, which in turn carries
increases
oxygen from the lungs to the tissues
o Contain a large quantity of carbonic anhydrase
o Enzyme that catalyzes the reversible
reaction between carbon dioxide (CO2)
and water to form carbonic acid (H2CO3)
o Transports carbon dioxide in the form of
bicarbonate ion from the tissues to the
lungs
o Responsible for most of the acid-base
buffering power of the whole blood
 Quantity of Hemoglobin in the Cell
o Hemoglobin
o Female – 14 grams Hgb/100 ml
o Male – 15 grams Hgb/100 ml
o Each gram of hemoglobin combines
with 1.34 ml of O2
o Hematocrit
𝑃𝑙𝑎𝑠𝑚𝑎 𝑣𝑜𝑙𝑢𝑚𝑒
o 𝑇𝑜𝑡𝑎𝑙 𝑏𝑙𝑜𝑜𝑑 𝑣𝑜𝑙𝑢𝑚𝑒 =
1−𝐻𝑒𝑚𝑎𝑡𝑜𝑐𝑟𝑖𝑡
o 40-45%

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PHYSIOLOGY Physiology of Red Blood Cells

A. GENESIS OF RBC B. REGULATION OF RBC PRODUCTION

 The blood begins their lives in the bone marrow

from a single type of cell, wherein all the cells of
 The functional capacity of the erythron,
the circulating blood are eventually derived =
therefore, requires normal renal production of
pluripotential hematopoietic stem cell
EPO, a functioning erythroid marrow, and an
 Differentiates to become Committed stem cells
adequate supply of substrates for hemoglobin
 CFU-erythrocytes – committed stem cells that synthesis.
produces erythrocytes
 Tissue oxygenation is the most essential
 First cell that can be identified to the RBC series regulator of RBC production
proerythroblast,
 Any condition that causes the quantity of oxygen
 The first-generation cells - basophil erythroblasts transported to the tissues to decrease ordinarily
-stain with basic dyes; the cell at this time has increases the RBC production:
accumulated very little hemoglobin.
 Anemia
o Polychromatophilic erythroblast: more
 Low blood volume
hemoglobin is being produced by the
mitochondria and the nucleus starts to condense  Low hemoglobin
o Orthochromatophilic erythroblast: the cell is  Poor blood flow
filled with hgb, most organelles are excreted and  Pulmonary disease
the pyknotic nucleus is ejected to become a  High altitude
reticulocyte
 EPO is the principal stimulus for RBC production
o Reticulocyte: it passes around the bloodstream
in low oxygen states
for 1-2 days to secrete any other remaining
organelles, Golgi apparatus and mitochondria
by diapedesis (squeezing through the pores of
the membrane) and then it becomes a mature
erythrocyte
 Because of the short life of the reticulocytes,
their concentration among all the RBCs is
normally slightly less than 1 percent
 Growth & reproduction of different stem cells are
controlled by multiple proteins called growth
inducers
o Major growth inducers:
o IL-3 – promotes growth & reproduction of
virtually all different types of committed stem
cells
o IL – 5 – promote growth of specific cells
o IL-6
o GM-CSF
o EPO
o Differentiation inducers
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PHYSIOLOGY Physiology of Red Blood Cells
C. STIMULATION FOR EPO PRODUCTION
 EPO formation increases in response to hypoxia
 Hypoxia causes a marked increase in
erythropoietin production
 90% of erythropoietin (EPO) is formed in the
kidneys, the remainder is formed in the liver
 In the absence of EPO, few red blood cells are
formed by the bone marrow
a. Function of EPO
 Erythropoietin stimulates red cell production by:
 promoting proerythroblasts production
 Shortening the transition time for the different
erythroblastic stages
 EPO is a glycoprotein hormone. It is produce and
release by the peritubular capillary lining cells of
the kidney. The fundamental stimulus for EPO
production is the availability of O2 for tissue
metabolic needs and key for EPO gene
regulation is the HIF-1 in the presence of O2.
HIF-1 is hydroxylated at a key proline allowing
your HIF-1 to be degraded by the proteasome D. VITAMIN B AND FOLIC ACID
pathway. If O2 becomes limiting this critical
 Requirement for RBC maturation:
hydroxylation step does not occur.
 Expression of the erythropoietin gene (EPO) is
inhibited by GATA-2 and NFκB, and stimulated
 Role of Vitamin B12 & Folate
by HIF-1. HIF-1 induces EPO expression under
hypoxic conditions, but degrades under
normoxic conditions.
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 HIF translocates to the nucleus, binds with
Hypoxia response element in the EPO gene 
induce transcription mRNA  upregulate
production of EPO
 Erythropoietin binds to the erythropoietin
receptors on bone marrow cells, stimulating the
production of red blood cells. In hypoxic state,
EPO is formed within minutes to hours and
mature RBCs appear after 5 days. EPO Red
blood cell production might be increased by the
following interventions: inhibition of PH, GATA-2
or NFκB, alteration of EPO–EPO receptor
interactions, stimulation of the EPO receptor, or
by making the EPO gene autonomous.
 Abbreviations: EPO, erythropoietin; HIF-1,
hypoxia-inducible transcription factor-1; NFκB,
nuclear factor κB; PH, prolyl hydroxylase.
 Low O2  EPO formed within minutes to hours
(max of 24h)  RBCs appears after 5 days
 EPO stimulate the formation of proerythroblasts
and its rapid differentiation through the different
erythroblastic stages => increase production of
new RBCs
12
o Vitamin B12 (cyanocobalamin)
o Folic acid (pteroylglutamic acid)
o Two reactions in the body that requires cobalamin
1. Methylation of homocysteine to methionine –
requiring methylcobalamin and 5-MTHF
 This reaction is the first step in the
pathway by which 5-MTHF, w/c enters
bone marrow and other cells from
plasma, is converted into all intracellular
folate co-enzymes
2. L-Methylmalonyl CoA isomerization which
requires deoxyadenosylcobalamin
(adocobalamin)
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PHYSIOLOGY Physiology of Red Blood Cells

 Role of intrinsic factor in the absorption of Vitamin B12

 Absorption of Folic Acid

o Unaltered folic acid is absorb in the proximal

jejunum where all the glutamyl residues is hydrolyse
by alpha 1 glutamyl transferase in the brush border
o Both vitamins in different pathways requires the
to form 5-methyl-THF this will be distributed in the
formation of thymidine triphosphate for DNA
body and converted to THF (tetrahydrofolic)
synthesis

 Vitamin B12 & Folic acid deficiency

o Causes abnormal & diminished DNA
 Failure of nuclear maturation & cell division 
fail to proliferate rapidly
o Produce larger RBCs (macrocytes)
 Irregular, & oval shaped
 Flimsy membrane

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PHYSIOLOGY Physiology of Red Blood Cells

III: ANEMIA  Heme is synthesized in a complex series of steps

A. CAUSES OF MEGALOBLASTIC ANEMIA involving enzymes in the mitochondrion and in the
cytosol of the cell. The first step in heme synthesis takes
COBALAMIN DEFICIENCY place in the mitochondrion, with the condensation of
Inadequate intake Vegan (rare)
Malabsorption A. Defective release of cobalamin from food succinyl CoA and glycine by ALA synthase to form 5-
 Gastric achlorhydria aminolevulinic acid (ALA). This molecule is transported
 Partial gastrectomy to the cytosol where a series of reactions produce a ring
 Drugs that block acid secretion
B. Inadequate production of intrinsic factor structure called coproporphyrinogen III. This molecule
 Pernicious anemia returns to the mitochondrion where an addition reaction
 Total gastrectomy
produces protoporhyrin IX. The enzyme ferrochelatase
 Congenital absence or function abnormality
of IF inserts iron into the ring structure of protoporphyrin IX to
C. Disorder of terminal ileum produce heme. Heme combines with long polypeptide
 Tropical sprue, Non-tropical sprue, regional
enteritis, intestinal resection, neoplasms globin to form hemoglobin chain ( or ).
and granulomatous disorders, Selective  Two distinct globin chains (each with its individual heme
cobalamin malabsorption
D. Competition for cobalamin molecule) combine to form hemoglobin. One of the
 Fish tapeworm chains is designated alpha. The second chain is called
 Bacteria: “blind loop” syndrome
E. Drugs: ASA, colchicine, neomycin
"non-alpha".
Others A. Nitrous oxide
B. Transcobalamin II deficiency
C. Congenital enzyme defects (rare)

FOLIC ACID DEFICIENCY

Inadequate intake Unbalanced diet (alcoholics, teenagers, infants)
Increased Pregnancy
requirements Infancy
Malignancy
Increased hematopoiesis (chronic hemolytic
anemias)
Chronic exfoliative skin disorders
Hemodialysis
Malabsorption Tropical sprue
Non-tropical sprue
Drugs: phenytoin, barbiturates
Impaired Inhibitors of dihydrofolate reductase:
metabolism methotrexate, pyrimethamine, triamterene, B. GLOBIN SYNTHESIS
pentamidine, trimethoprim
Alcohol
Embryonic HGB Fetal HGB Adult HGB
Rare enzyme deficiencies: dihydrofolate
reductase Gower 1 –  (2) –  (2) Hgb A –  (2),  (2)
Gower 2 –  (2),  (2) Hgb F –  (2),  (2) Hgb A2 –  (2),  (2)
Portland –  (2),  (2)
*alpha – , beta – , delta – , epsilon – , zeta - 
IV: HEMOGLOBIN
A. HEMOGLOBIN FORMATION  With the exception of the very first weeks of
 Synthesis of hemoglobin begins in the proerythroblast embryogenesis, one of the globin chains is always
to the reticulocyte stage alpha. The fetus has a distinct non-alpha chain called
 Chemical steps in the formation of hemoglobin gamma. After birth, a different non-alpha globin chain,
called beta, pairs with the alpha chain. The combination
of two alpha chains and two non-alpha chains produces
a complete hemoglobin molecule. The combination of
two alpha chains and two gamma chains form "fetal"
hemoglobin, termed "hemoglobin F". With the
exception of the first 10 to 12 weeks after conception,
fetal hemoglobin is the primary hemoglobin in the
developing fetus. The combination of two alpha chains
and two beta chains form "adult" hemoglobin, also
called "hemoglobin A“ & it becomes the predominate
hemoglobin within about 18 to 24 weeks of birth.

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PHYSIOLOGY Physiology of Red Blood Cells

 The genes that encode the alpha globin chains are on
chromosome 16. Those that encode the non-alpha
globin chains are on chromosome 11.
C. HEMOGLOBIN MOLECULE
 The pairing of one alpha chain and one non-alpha chain
produces a hemoglobin dimer (two chains). The
hemoglobin dimer does not efficiently deliver oxygen.
Two dimers combine to form a hemoglobin tetramer,
which is the functional form of hemoglobin, which
control oxygen uptake in the lungs and release in the
tissues that is necessary to sustain life.
a. Structure of heme moiety
2. Molecular compounds are made due to covalent
bonding while ionic compounds are made due to
ionic bonding.
3. Molecular compounds are formed between two
non-metals while ionic compounds are formed
between metals and non-metals.
4. Molecular compounds are poor electrical
conductors while ionic compounds are good
conductors.
5. Molecular compounds can be in any physical
state ‘“solid, liquid, or gas. Ionic compounds are
always solid and crystalline in appearance.
6. There are a lot of molecular compounds than ionic
compounds.

V: IRON
a. Quantity of Iron in the Body
 Total iron in the body – 4-5 g
 65% in the form of haemoglobin
 4% - myoglobin
 1% - various heme compounds
 0.1 % - with transferrin in the blood plasma
 15-30% - stored iron
b. Absorption, Transport & Storage

 Under normal conditions, the iron is in the ferrous

(Fe2+) oxidation state. It lies slightly outside the plane
of the porphyrin in heme. The iron ion can form two
additional bonds, one on each side of the heme plane.
These binding sites are called the fifth and sixth
coordination sites.
b. Position of Iron in Deoxyhemoglobin
 In hemoglobin, the fifth coordination site is occupied
by the imidazole ring of a histidine residue from the
protein. In deoxyhemoglobin, the sixth coordination
site remains unoccupied.
 In oxyhemoglobin, the iron ion moves into the plane
of the heme on oxygenation. The proximal histidine is
pulled along with the iron ion.
c. Hemoglobin combines loosely & reversibly  Iron is absorbed in the SI (duodenum). The liver
with oxygen secretes apotransferrin into the bile into the
 This is the most important feature of the Hgb duodenum & combines with iron to form transferrin,
molecule. which in turn binds with the receptors in the intestinal
 O2 molecule binds loosely with one of the so called epithelial cell, enters the cell through pinocytosis then
coordination bonds of the iron atom, the combination released to the blood stream in the form of plasma
is easily reversible transferrin. Macrophages release iron from the Hgb &
1. Molecular compounds are pure substances goes back to the blood with transferrin  goes to
formed when atoms are linked together by tissues or stored as ferritin
sharing of electrons while ionic compounds are  HCP1 – heme carrier protein
formed due to the transfer of electrons.  DMT1 – divalent metal transporter 1
 FP – ferroportin
 TfR – transferrin receptor
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PHYSIOLOGY Physiology of Red Blood Cells

c. Iron Transport & Metabolism

 When there is IDA RBC have too little HGB and they
look smaller and paler. They will present as microcytic
and hypochromic anemia

References:
 Dr. Remudaro’s RBC Powerpoint
 Dr. Remudaro’s RBC Video lecture
 Hall, J. E. (2016). Guyton and Hall textbook of
medical physiology (13th ed.) Philadelphia, PA:
Elsevier

 Total iron in the body – 4-5 g

 The total body iron is regulated by altering the rate of
absorption. When the amount of iron in the body is
more than the apoferritin storage pool, iron is
converted into an insoluble form called hemosiderin
(iron supplements). Excess iron is specially deposited
in the hepatocytes and less in the bone marrow.
When the apoferritin are saturated with iron the rate
of absorption decreases and conversely increases
>5x when the stores are depleted like in blood loss
 Destruction of hemoglobin
 When red blood cells burst and release their
hemoglobin
 The hemoglobin is phagocytized by macrophages in
many parts of the body
 Macrophages release iron from the hemoglobin and
reused for production of new RBCs
 RBC membrane becomes fragile & they rupture as
they pass through though tight areas in the circulation
or when they self-destruct in the spleen

NUTRITIONAL IRON BALANCE

Route by Mechanisms by which iron is lost from the
which iron body
comes into
the body
Absorption Increased demand for iron and/or
from food or hematopoiesis
from  Rapid growth in infancy & adolescence
medicinal iron  Pregnancy
 Erythropoietin therapy
Red cell Increased iron loss
transfusion  Chronic or acute blood loss
 Menses
 Blood donation
 Phlebotomy as treatment for polycythemia
vera
Injection of Decreased iron intake or absorption
iron  Inadequate diet
complexes  Malabsorption from disease (sprue,
Crohn’s disease)
 Malabsorption from surgery
 Acute or chronic inflammation

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