BIOL380

LECTURE 2
• Erythropoiesis:
• Production of red blood cells
• Approximate 1012 made daily
• Process:
Stem cell

CFUGEMM (progenitor cells colony

forming unit granulocyte, erythroid,
monocyte, megakaryocyte

BFUE (burst forming unit erythroid)

CFUE (erythroid colony forming

unit), pronormoblast

pronormoblast (first recognizable

erythrocyte precursor)
• Pronormoblast:
• First recognizable
erythrocyte
precursor
• Large cell
containing dark
blue cytoplasm,
central nucleus
with nucleoli and
slightly clumped
chromatin
• Gives rise to
smaller
normoblasts
(nucleated rbc) via
cell division
• As a result of
divisions, more
haemoglobin is
formed in
cytoplasm
• Pronormoblast:
• Cytoplasm
becomes paler
blue to pink as
RNA and protein
synthetic
apparatus
(ribosomes) are
lost
• Nucleus is finally
extruded from late
normoblast
• This occurs within
the marrow
• Now referred
to as a
reticulocyte
(circulates in
the blood for 1
– 2 days)
• Still contains
rRNA thus able
to synthesize
haemoglobin
• 16 mature rbcs
produced from
1
pronormoblast
• Normoblast:
• Not present in normal human
peripheral blood
• Appears during extramedullary
erythropoiesis/marrow diseases

• Reticulocyte:
• Slightly larger than mature red
blood cell
• Spends 1 – 2 days in bone marrow
• Circulates 1 – 2 days in peripheral
blood before maturing (RNA is then
completely lost)
• Mature rbc is pink-staining, non-
nucleated and biconcave
 Erythropoietin
• 90 % of erythropoietin
hormone is produced in the
peritubular interstitial cells
(endothelial cells of the cortex
and the outer medulla) of the
kidney
• 10% produced by liver
• Regulates erythropoiesis
• Erytropoitin
• Production is stimulated by oxygen
tension in tissues of kidney
• Hypoxia induced HIF- 2α and β
factors (hypoxia-inducible factors)
stimulate Epo production
• Stimulates erythropoiesis by
increasing the number of
progenitor cells that are committed
to erythropoiesis (N.B. take note of
the transcription factors activated
by Epo receptor stimulation p. 18)
Erytropoitin
• Stimulates
erythropoiesis by
increasing the
number of
progenitor cells that
are committed to
erythropoiesis (N.B.
take note of the
transcription factors
activated by Epo
receptor stimulation
p. 18)
• Factors which increase Epo
production:
• In anemia
• When haemoglobin is metabolically or structurally unable to
give up oxygen normally
• When atmospheric oxygen is low
• When defective cardiac/pulmonary function/damage to
renal circulation affects oxygen delivery to kidney
• Precursors for effective erythropoiesis:
• Metals – Iron, cobalt
• Vitamins – B12, folate, C, E, B6, thiamine, riboflavin
• Hormones – androgens, thyroxine

N.B. A deficiency in any of these may be associated with

anaemia (a condition in which there is a deficiency of
red cells or of haemoglobin in the blood, resulting in
pallor and weariness)
 Haemoglobin
• Haemoglobin is needed for rbcs to transport oxygen to tissues and return carbon
dioxide from tissues to lungs
• Each rbc contains approx. 640 million haemoglobin molecules
• Each molecule of normal haemoglobin A (Hb A) (dominant haemoglobin)
consists of 4 polypeptide chains, α2β2, each having its own haem group.
• Other haemoglobins: Hb F (fetal)and Hb A2 (second type of Hb found in adults in small amounts)
• Major switch from fetal to adult haemoglobin 3 – 6 months after birth
• Synthesis occurs in mitochondria
 Haemoglobin synthesis
• Glycine and succinyl coenzyme A plus vitamin B6
(pyridoxal phosphate/pyridoxine) are required
• Enzyme needed for reaction is delta aminolaevulinic acid
(ALA) synthase
• Reaction is stimulated by Epo
• This results in the synthesis of protoporphyrin which then
binds with Fe2+to form haem
• A molecule of haem then binds with a globin chain (made
on polyribosomes)
• 4 globin chains each having a haem group in a “pocket”
makes up a haemoglobin molecule
 Function of Haemoglobin
• Oxygen is loaded and
unloaded onto haemoglobin
molecule
• During these processes,
globin chains move on each
other
• Alpha1beta1 and
alpha2beta2 contacts
stabilize the molecule
 Function of Haemoglobin
• Beta chains slide on contacts during oxygenation and deoxygenation
• When oxygen is unloaded, beta chains are pulled apart, allowing 2,3
diphosphoglycerate (2,3-DPG) resulting in lower affinity for oxygen
 Function of Haemoglobin
• This gives sigmoid form of
haemoglobin oxygen dissociation
curve.
• P50 (partial pressure of oxygen at
which haemoglobin is half saturated
with oxygen) of normal blood is 26
mmHg
• In vivo, O2 operates between 95 %
saturation (arterial blood) with a
mean arterial O2 tension of 95
mmHg and 70% saturation (venous
blood) with a mean venous tension
of 40 mmHg
 Clinical Application
• Methaemogoblinemia:
• Clinical state in which in circulating hemoglobin iron is in its 3+ state
instead of its 2+ state
• Arises because of hereditary deficiency of methaemoglobin
reductase/inheritance of structurally abnormal haemoglobin (Hb M)
• HbM has an amino acid substitution which affects haem pocket of
globin chain
 Clinical Application
• Toxic methaemoglobinaemia occurs when drugs/toxic substances
oxidize haemoglobin
• Patient shows cyanosis

https://medizzy.com/feed/1260188?title=methemoglobinemia
 Red Blood Cell
• RBCs are necessary for gaseous exchange and must bring Hb in close
contact with tissues.
• As a result, RBCs which are 8 micrometers in diameter:
• Must pass through vessels that are 3.5 micrometers in diameter
• Must maintain haemoglobin in ferrous state
• Must main osmotic equilibrium in spite of high protein concentration
(haemoglobin)
 Red Blood Cell
• All of the above is possible because of
the following:
• shape of cell (biconcave disc which is
flexible)
• Ability to generate ATP by anaerobic
glycolytic pathway (Embden-
Meyerhof pathway)
• Generate reducing power as NADH
by E-M pathway
• And as reduced NADPH by the
hexose monophosphate shunt
 RED CELL METABOLISM
• Embden-Meyerhof glycolytic
pathway:
• Glucose enters cells via
facilitated transfer. End product
is lactate
• One molecule of glucose gives 2
molecules of ATP
• This ATP provides energy for
maintenance of red cell volume,
shape and flexibility
 RED CELL METABOLISM
• Membrane become weakened because of continual Na+ and K+
movement
• Membrane ATPase sodium pump uses one molecule of ATP to move
3 Na+ ions out of and 1 K+ ion into the cell
• Generates NADH needed by methaemoglin reductase to reduce
functionally dead methaemoglobin (oxidized hemoglobin)
containing Fe3+ to functionally active reduced haemoglobin
 Hexose monophosphate pathway
• Approximately 10% of glycolysis
occurs using this pathway
• Glucose to glucose-6-phosphate to 6-
phosphogluconate to ribulose-5-
phosphate
• NADPH is generated
• Linked with gluthathione (oxidized) to
maintain haemoglobin and RBC
membranes
• Used by methaemoglobin reductase
to maintain haemoglobin in its active
Fe2+ state
 RED CELL MEMBRANE
• Lipid bilayer, integral membrane
proteins, membrane skeleton
• Proteins (peripheral/integral) – 50 %;
Phospholipids – 20%; cholesterol
molecules – 20%; 10 % carbohydrates
(external surface only)
• Structural proteins: α and β spectrin,
ankyrin, protein 4.1 and actin
• These form a horizontal lattice on
internal side and maintain biconcave
shape
• Most abundant is spectrin; made up of
2 chains, α and β wound around each
other forming heterodimers
 ANAEMIA
• Defined as a reduction in haemoglobin concentration of blood
below normal for age and sex
• Reduction in haemoglobin is usually accompanied by a fall in
rbc count and packed cell volume (PCV)
• Alterations in total circulating plasma volume plus total
circulating haemoglobin mass determine haemoglobin
concentration
• 1. (Hb) Hemoglobin - The amount of Hb in the blood, in g/dl. (Low is
called anemia.)
2. (HCT) Hematocrit - Fraction of whole blood volume that consists of
red cells.
3. (RBC) Total red blood cells - Number of red cells given as an absolute
number of cells/liter.
4. (MCH) Mean corpuscular hemoglobin - the average hemoglobin per
red cell, in picograms.
5. (MCV) Mean corpuscular volume - the average volume of the red
cells, measured in femtolitres. Anemia is classified as microcytic or
macrocytic based on whether this value is above or below the
expected normal range
6. (MCHC) Mean corpuscular hemoglobin concentration - the average
concentration of Hb/cell.
 Clinical Features of Anaemia
The following are used to consider the clinical features of anaemia
•Speed of onset
• More symptoms seen with rapidly progressive anaemia than slow onset because of less
time for adaption by CV system and in oxygen dissociation curve of haemoglobin
•Severity
• Mild anaemia often produces no symptoms/signs. Symptoms often present when
haemoglobin levels are less than 9 – 10 mg/dl
•Age
• Anaemia poses more problems in the aged because of reduced oxygen and its effects on
organs
•Haemoglobin dissociation curve
• Anaemia is associated with a rise in 2,3 DPG in red cells and a shift in O2 dissociation curve
to the right so that oxygen is given up more readily to tissues
 Symptoms
• Symptoms:
• Shortness of breath
• Weakness
• Lethargy
• Palpitation
• Headaches
• Symptoms of cardiac failure; angina pectoris, intermittent claudication (in older
individuals)
• Visual disturbances
 SIGNS
• May be general of specific
• General:
• Pallor of mucous membrane if haemoglobin levels are less than 9
– 10 mg/dl
 SIGNS
• Specific:
• Those associated with particular types.
• Koilonychia (spoon nails/washboard nails) with iron deficiency

• https://glutenfreeworks.com/health/koilonychia-3/
 SIGNS
• Specific:
• Those associated with particular types.
• Jaundice with haemolytic/megaloblastic anaemia

• https://www.msdmanuals.com/home/quick-facts-liver-and-gallbladder-disorders/manifestations-of-liver-
disease/jaundice-in-adults
 SIGNS
• Specific:
• Leg ulcers with sickle cell anemia

https://onlinelibrary.wiley.com/doi/full/10.1002/ajh.24134
 SIGNS
• Specific:
• Those associated with particular types.
• Bone deformities with thalassemia major

https://naturereview.wordpress.com/2014/04/08/
types-of-thalassemia/

https://www.joms.org/article/S0278-2391(11)01832-5/references
 SIGNS
• Specific:
• Those associated with particular types.
• Spontaneous bruising/anaemia with
infections as a result of
neutropenia/thrombocytopenia

https://www.msdmanuals.com/professional/hematology-and-oncology/ https://www.msdmanuals.com/professional/hematology-and-oncolo
thrombocytopenia-and-platelet-dysfunction/immune-thrombocytopenia-itp thrombocytopenia-and-platelet-dysfunction/immune-thrombocytope
Classification of Lab. findings
 Classification of Lab. findings
• Blood films should be
examined