APOPTOSIS

Apo: apart
Ptosis: fallen
Shedding of leaves from tree
 INTRODUCTION

Cell death by injury

-Mechanical damage
-Exposure to toxic chemicals
Cell death by suicide
-Internal signals
-External signals
Conted…..

Apoptosis or programmed cell death, is carefully

coordinated collapse of cell, protein degradation ,
DNA fragmentation followed by rapid engulfment
by phagocytic cells.

Essential part of life for every multicellular

organism from worms to humans.
Apoptosis plays a major role in embryonic
development.
Why should a cell commit suicide?
Apoptosis is needed for proper development
Examples:
– The resorption of the tadpole tail
– The formation of the fingers and toes of the fetus
– The formation of the proper connections between neurons in the brain

Apoptosis is needed to destroy cells

Examples:
– Cells infected with viruses
– Cells of the immune system
– Cells with DNA damage
– Cancer cells
 APOPTOSIS: important in embryogenesis

Morphogenesis (eliminates excess cells):

Selection (eliminates non-functional cells):

 APOPTOSIS: important in embryogenesis

Immunity (eliminates dangerous cells):

Self antigen
recognizing cell

Organ size (eliminates excess cells):

 APOPTOSIS: important in adults
Tissue remodeling (eliminates cells no longer needed):

Apoptosis
Virgin mammary gland Late pregnancy, lactation Involution
(non-pregnant, non-lactating)

- Testosterone
Apoptosis

Prostate gland
 APOPTOSIS: important in adults
Tissue remodeling (eliminates cells no longer needed):

Apoptosis

Resting lymphocytes + antigen (e.g. infection) - antigen (e.g. recovery)

Steroid immunosuppressants: kill

lymphocytes by apoptosis

Lymphocytes poised to die by apoptosis

 APOPTOSIS: important in adults
Maintains organ size and function:

Apoptosis
X
+ cell division

Cells lost by apoptosis are replaced by cell division

What makes a cell decide to commit suicide?

Withdrawal of positive signals

examples :
– growth factors for neurons

Receipt of negative signals

examples :
– increased levels of oxidants within the cell
– damage to DNA by oxidants
– death activators :
• Tumor necrosis factor alpha (TNF-)
• Lymphotoxin (TNF-β)
• Fas ligand (FasL)
STAGES OF CLASSIC APOPTOSIS
Healthy cell

DEATH SIGNAL (extrinsic or intrinsic)

Commitment to die (reversible)

EXECUTION (irreversible)

Dead cell (condensed, crosslinked)

ENGULFMENT (macrophages, neighboring cells)

DEGRADATION
Cells are balanced between life and death
DAMAGE Physiological death signals

DEATH SIGNAL

PROAPOPTOTIC ANTIAPOPTOTIC
PROTEINS PROTEINS
(dozens!) (dozens!)

DEATH
 Caspases
• Proteins which degrade other proteins
are employed by apoptosis - caspases
• Made as inactive precursors -
procaspases
• These are activated by other proteins
when the right signal is received
• One caspase cleaves the lamin proteins
resulting in the irreversible breakdown
of the nuclear membrane.
 APOPTOSIS: control
Receptor pathway (physiological):
FAS ligand TNF

Death receptors:
(FAS, TNF-R, etc)
Death
domains

Adaptor proteins

Pro-caspase 8 (inactive) Caspase 8 (active)

Pro-execution caspase (inactive)

Execution caspase (active)

MITOCHONDRIA Death
 APOPTOSIS: control
Intrinsic pathway (damage):

Mitochondria

BAX Cytochrome c release BCL-2

BAK BCL-XL
BOK BCL-W
BCL-Xs Pro-caspase 9 cleavage MCL1
BAD BFL1
BID DIVA
B IK NR-13
BIM Pro-execution caspase (3) cleavage Several
NIP3 viral
BNIP3 proteins

Caspase (3) cleavage of cellular proteins,

nuclease activation,
etc.

Death
 Apoptosis: Pathways
“Extrinsic Pathway”

Death Death Initiator

Ligands Receptors Caspase 8

Effector
“Intrinsic Pathway”
Caspase 3 PCD

DNA Initiator
Mitochondria/
damage Caspase 9
Cytochrome C
& p53
 APOPTOSIS: control

Physiological Intrinsic
receptor pathway damage pathway

MITOCHONDRIAL SIGNALS

Caspase cleavage cascade

Orderly cleavage of proteins and DNA

ENGULFMENT
(no inflammation)
 APOPTOSIS: Morphology

organelle
reduction
membrane
blebbing &
changes
cell
mitochondrial
leakage shrinkage

nuclear chromatin
fragmentation condensation

Hacker., 2000
 APOPTOSIS: Morphological events
cell shrinkage
organelle reduction
mitochondrial leakage
chromatin condensation
nuclear fragmentation
membrane blebbing & changes
Blebbing & Apoptotic bodies
The control retained over the cell
Bleb membrane & cytoskeleton allows intact
pieces of the cell to separate for
recognition & phagocytosis by MFs

Apoptotic body

MF MF
 Apoptosis signal to kill infected cells
Cytolytic lymphocyte/CTL (& natural killer lymphocyte)
presents Fas ligand/CD178 on its surface to tell the infected
cell to die

Fas ligand

Externally driven
Apoptotic signals
Cytochrome c

Initiator caspases
The immunological
synapse holds the cells
much tighter together
Execution caspases
than shown here

Apoptosis events
Fas/ CD95 is the
‘death receptor’
DNA DAMAGE

p53
APOPTOSIS: Role in Disease

TOO MUCH: Tissue atrophy

Neurodegeneration
Thin skin
etc

TOO LITTLE: Hyperplasia

Cancer
Athersclerosis
etc
 APOPTOSIS: Role in Disease
Neurodegeneration

Neurons are post-mitotic (cannot replace themselves;

neuronal stem cell replacement is inefficient)

Neuronal death caused by loss of proper connections,

loss of proper growth factors (e.g. NGF), and/or
damage (especially oxidative damage)

Neuronal dysfunction or damage results in loss of synapses

or loss of cell bodies

PARKINSON'S DISEASE
ALZHEIMER'S DISEASE
HUNTINGTON'S DISEASE etc.
 APOPTOSIS: Role in Disease
Cancer

Apoptosis eliminates damaged cells

(damage => mutations => cancer

Tumor suppressor p53 controls apoptosis responses to damage

Most cancer cells are defective in apoptotic response

(damaged, mutant cells survive)

High levels of anti-apoptotic proteins

or
Low levels of pro-apoptotic proteins
===> CANCER
OPTIMAL FUNCTION (HEALTH)

APOPTOSIS

APOPTOSIS
Neurodegeneration cancer, …..
 Necrosis vs. Apoptosis
Necrosis Apoptosis
• Cellular swelling • Cellular condensation
• Membranes are broken • Membranes remain intact
• ATP is depleted • Requires ATP
• Cell lyses, eliciting an • Cell is phagocytosed, no
inflammatory reaction tissue reaction
• DNA fragmentation is • Ladder-like DNA
random, or smeared fragmentation
• In vivo, whole areas of • In vivo, individual cells
the tissue are affected appear affected
NECROSIS Vs APOPTOSIS

Wilde, 1999