APOPTOSIS

DR.AYESHA FARHEEN
Objectives

Definition of Differentiation Applied

Process Pathways
Apoptosis from Necrosis Aspect
 APOPTOSIS
Greek word : Falling leaves (like in Autumn)

In the human body about 100,000 cells are produced every second by mitosis and a
similar number die by apoptosis !!!
History

 Apoptotic principle was first described by KARL VOGT in 1842

 “APOPTOSIS” term was coined by JAMES CORMACK
 2002 NOBEL PRIZE IN MEDICINE was awarded to SYDNEY
BRENNER,HORVITZ and JOHN.E.SULSTON for their work in identifying genes
that control Apoptosis.

John E Sulton won Nobel Prize in 2002

for his pioneering research in Apoptosis
PROGRAMMED CELL DEATH

CELL SUICIDE
What is Apoptosis?

 DEFINITION : Apoptosis is a peculiar well controlled individual cell death that is

caspase mediated and leads to fragmentation of the cell and organelles into
numerous small buds, which are then engulfed by macrophages without
surrounding inflammation

An orderly
disposal of cells
that need to die

Cell is just too DNA has

old and ’ its sustained too
time has come’ many injuries

Cell needs to be
removed for Cell is infected
body parts to with a virus
be formed
Most of the embryo development involves programmed cell death

The tail of a tadpole is absorbed via apoptosis.

Importance of Apoptosis

1) Crucial for embryonic development

-Errors in Apoptosis can lead to Birth Defects
2) Important for maintaining homeostasis
- Cell death is balanced with mitosis to regulate cell number.
3) Improper regulation contributes to human disease
- Neurodegenerative diseases
Parkinson’s
Alzheimer’s
- Cancer
- Autoimmune diseases e.g. (diabetes type I)
- Viral diseases
What is Apoptosis NOT?

 Apoptosis is NOT cell death after injury

Cell death after injury is called NECROSIS
Cells die by one of two mechanisms – necrosis or apoptosis
Two physiologically different processes
– Necrosis – death by injury
– Apoptosis – death by suicide
Apoptosis and necrosis have different characteristics
How Apoptosis Differs from Necrosis?

1. Apoptosis is intrinsically controlled, necrosis is not

2. Apoptosis is more rapid (12-24 hours) than necrosis
3. Apoptosis is induced by endogenous or exogenous stimuli, necrosis is always induced by
exogenous harms
4. Apoptosis is limited to single or few cells at a time, and occurs among healthy cell
population, necrosis is usually more extensive & occurs in tissue exposed to injuries
5. Cell cytoplasm shrinks in apoptosis and swells in necrosis.
6. Nucleosomes of apoptotic cells are 180 bp fragments, contrary to the irregular ones in
necrosis
7. Apoptosis has no inflammation, while necrosis leads to liberation of pro-inflammatory
mediators
8. Apoptosis has no systemic manifestations contrary to most inflammations
What happens during Apoptosis?

Nuclear
Nuclear fragmentation
condensation
Breakdown of
mitochondria;
release of
cytochrome C
Cell shrinkage
(condensation of
cytoplasm)

A programmed
series of events
occur
Cellular changes associated with apoptosis
 Biochemical features of Apoptosis

Protein • By activation of caspases

Cleavage • Caspases activate DNAses

• Cleavage into oligonucleosomes

DNA
• By Ca2+-and Mg2+-dependent
Breakdown endonucleases

Phagocytic • Phosphatidylserine
Recognition • Thrombospondin
Mechanisms of Apoptosis
 Mechanisms of Apoptosis

Active cysteine residue in the catalytic site

Specificity in cleavage after an Asp residue

Synthesized as inactive zymogens (PROCASPASES)

 Digestion of DNA starts after
2 hrs
 3&4 hrs after initiation of apoptosis DNA is almost all
degraded
 DNA is fragmented with restriction endonucleases
 Apoptosis induces 180 bp Laddering of DNA
• DNA cleaved into non-random fragments
• 180-200 bp fragments & multiples of this unit
Mechanism

Four stages of apoptosis have been defined:

i. Commitment to death by extracellular or intracellular triggers/signals

ii. Cell killing (execution) by activation of intracellular proteases (caspases)

iii. Engulfment of cell corpse by other cells

iv. Degradation of the cell corpse within the lysosomes of phagocytic cells
DAMAGE Physiological death signals

DEATH SIGNAL

PROAPOPTOTIC ANTIAPOPTOTIC
PROTEINS PROTEINS
 APOPTOSIS

MITOCHONDRIAL SIGNALS
Stimuli for Apoptotic Cell Death in Mammals
i. Growth factor deficiencies
ii. Ionizing radiation/ viral infection
iii. Free radical toxicity
iv. Death receptor activation (such as Fas or CD95 triggering)
v. Metabolic or cell cycle disturbance
Stimuli for Apoptotic Cell Death
Death Factors

Definition: Cytokines that activate an apoptosis program by binding to their

specific receptor. Typical examples of death factors are:
1. Fas ligand,
2. TNF (tumor necrosis factor) and
3. TRAIL (TNF-related apoptosis-inducing ligand).
- Apoptosis can also be induced by cytotoxic T-lymphocytes using the enzyme
granzyme.
 Apoptosis

Extrinsic Intrinsic Granzyme

pathway pathway pathway
Extrinsic pathway

Fas engagement. FADD= Fas associated

death domain.
Extrinsic pathway
Intrinsic Pathway

AIF= Apoptosis inhibitory factor; IAPs=

Inhibitors of apoptosis proteins; Apaf-
1= apoptosis protease activating factor
Intrinsic Pathway
 Bcl2 was the first apoptosis-related gene ,recognized to play a role tumor genesis

 BCL-2 is a human proto-oncogene located on chromosome 18.

 Its product is an integral membrane protein (called Bcl-2) located in the membranes of the endoplasmic
reticulum , nuclear envelope, and in the outer membrane of mitochondria.

 The gene was discovered as the translocated locus in a B-cell leukemia (hence the name). This
translocation is also found in some B-cell lymphomas
Apoptosis and Cancer
Apoptosis and Cancer

• Apoptosis does not occur in Cancer

• Cancerous cells trick and skip Apoptosis in number of ways
 Inactivation of p53 [shooting the guard]
 Produce Bcl-2 or a protein which mimics Bcl-2
 Inhibits expression of Apaf-1
Apoptosis and Autoimmune Disease

RBC
Hemolytic
Anemia

Autoimmune Apoptosis doesnot

Lymph Proliferative occur in self
Syndrome[ALPS] Neutrophil
reactive T & B cells Neutropenia

Platelets
Thrombocyto
penia
Apoptosis and HIV

Infected CD4 • Deactivated

Infected CD4 cell
cell induces Bcl-2
avoids Apoptosis in
Apoptosis in • Decreases
itself
surrounding CD4
Decreases
cells Glycoprotein
Phosphatdylserine
markers on
marker for itself
innocent T
allowing longer
cells ,getting
survival.
them killed
 Overview

Apoptosis is a good thing

Too little of a good thing is
bad [Cancer]
Too much of a good thing is
also bad [HIV]
Sources

 Textbook of Medical Physiology –Guyton & Hall [12th edition]

 Review of Medical Physiology-William F Ganong [24th edition]
 Internet Sources :
• www.ncbi.nlm.nih.gov/books/NBK26873
• http://en.wikipedia.org/wiki/Apoptosis
Articles :
Apoptosis- Molecular mechanisms and Pathogenicity
http://link.springer.com/article/10.1023/A:1009616228304#pg2
http://www.excli.de/vol8/Rastogi_08_2009/Rastogi_030809_proof.pdf
“
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THANK YOU