FOUNDATIONS: The Embryo

Apoptosis

Dr. John Th’ng

GC11
jthng@auamed.net
“Life is a sexually transmitted
ultimately fatal disorder”
FO1.26: Understand the process of
programmed cell death (apoptosis) in the
development of an embryo
FO1.26.1: Recognize the major forms of cell death
and their characteristics under normal physiological
conditions
FO1.26.2: Identify the major enzymes involved in
the degradation of cellular components during
apoptosis
FO1.26.3. Describe the signalling pathways that
regulate apoptosis and the consequences of
uncontrolled apoptosis in health and disease
Cell Suicide vs. Cell Murder
- why do some cells die while others live?
- what mechanisms regulate cell survival?
- 2 different ways to turn on cell death
- absence of trophic (survival) factors
- cell “suicide”
- kill signals turn on cell death
-“murder” of target cells
Cell Suicide vs. Cell Murder

Cellular response depends on specific combination of signals

Cell death
- A common biological event
- Major pathways
- necrosis
- necroptosis
- apoptosis
Cell death
Necrosis
- rapid process of cell destruction
- cell membrane rupture
- caused by trauma, infections, other
external forces
- ischemia, gangrene, heat, etc.
- leakage of cellular contents
- can lead to inflammatory and
immunological responses
Cell death
Necrosis
- Classified by morphology:
- coagulative, liquefactive, gangrenous,
caseous, fat, fibrinoid
Necroptosis
- Necrosis that is induced in a programmed
manner
- example: defence against viruses
- No activation of caspases
- Triggers permeabilization of cell membrane
Cell death pathways - apoptosis
- normal process to balance cell growth
- present in all cells
- remove damaged, excess, or “old” cells
- involve programmed sequence of
molecular events: programmed cell
death, or apoptosis
- responsible for majority of cell death
- cells become shrivelled, but remain intact
- do not release cellular contents
Cell death pathways

Figure 23.1 Lippincott

Cell & Molecular
Biology
 Apoptotic cells Necrotic cell

Figure 18–1 Molecular Biology of the Cell

Apoptosis: Programmed Cell Death

SEM: normal (L) and apoptotic (R) T-cell

TEM: apoptotic cell

with membrane
blebbing
Roles of Apoptosis
Embryonic development:
• form structure, organs and tissues
• deletion of interdigital tissue in digit formation
• pruning unneeded nerve cells in nervous system
Adult tissues:
• balances cell proliferation to maintain organ sizes
• maintain constant cell numbers in tissues
- eg. bone marrow
• prevents production of new virus particles and limits
spread of virus
• eliminate cells carrying potentially harmful mutations and
prevent cancers, DNA damage
Roles of Apoptosis
Maintain homeostasis:

[bcl2]
Roles of Apoptosis

Figure 23.3 Lippincott Cell

& Molecular Biology
Roles of Apoptosis

Figure 18–2 Molecular Biology of the Cell

Apoptosis
- determine shape and size of limbs
and tissues during development
- sculpt the features of hands and fingers
- more than half the cells die during
nervous system development
- removal of lymphocytes after
infections
FO1.26: Understand the process of
programmed cell death (apoptosis) in the
development of an embryo
FO1.26.2: Identify the major enzymes involved in
the degradation of cellular components during
apoptosis
Apoptosis
- triggered by specialized proteases:
caspases
- requires activation by signals
- initiator caspases cleaves and activate
the executioner caspase to catalyze
cellular proteolysis
- nuclear lamina, cytoskeleton, cell adhesion
molecules, nuclease activation, etc.
- DNA and chromatin degradation
- cells shrivel up
Apoptosis
- triggered by specialized proteases:
caspases
Apoptosis

Figure 18–3 Molecular Biology of the Cell

Apoptosis
Caspase activate endonuclease to cleave
DNA in the nucleus
Caspase
activated
DNase

Figure 18–4 Molecular Biology of the Cell

FO1.26: Understand the process of
programmed cell death (apoptosis) in the
development of an embryo
FO1.26.3. Describe the signalling pathways that
regulate apoptosis and the consequences of
uncontrolled apoptosis in health and disease
- how is it triggered
- how is it inhibited
Apoptosis
- important for homeostasis
- highly regulated process in cells
- activated only when needed
- can cause diseases if uncontrolled
- too much can cause Alzheimer’s,
Parkinson’s, ALS, etc.
- too little can cause cancers, autoimmune
diseases, etc.
Apoptosis
How is it triggered?
- extrinsic pathway
- triggered by external signals
- intrinsic pathway
- triggered by internal signals
Apoptosis
How is it triggered?
- extrinsic pathway
- external signals bind cell-surface death
receptors
- tumor necrosis factor (TNF) receptor family
- only those that contain a death domain
- eg. Fas ligand (FasL) in lymphocytes
- internal death domain activate apoptosis
- death-inducing signaling complex (DISC)
- activate initiator caspase
- activate executioner caspase
 Apoptosis
Extrinsic pathway:

Figure 18–5 The extrinsic pathway

of apoptosis activated through Fas
death receptors.
Apoptosis
Extrinsic pathway:

Figure 18–5 The extrinsic pathway of apoptosis activated through

Fas death receptors.
Apoptosis
Extrinsic pathway
- regulated by inhibitor protein c-FLIP
- dimerizes with initiator caspase-8, but
has no caspase activity
- inactivates executioner caspase
Apoptosis
How is it triggered?
- intrinsic pathway
- internal signals: developmental signals or
cell stress, eg. DNA damages
- mitochondrial pathway that releases
mitochondrial proteins (cytochrome C)
- binds protein Apaf1 (apoptosis protease
activating factor) to form apoptosome
- activate initiator caspase to activate executioner
caspase
Apoptosis
Intrinsic pathway

Figure 18–7 The intrinsic pathway of apoptosis

Apoptosis
Intrinsic pathway
- tightly regulated by Bcl2 protein family
- members are either pro- or anti-apoptotic
- members can bind to inhibit each other
- balance determines life or death of cell
- anti-apoptotic: Bcl2 and BclXL
- block release of cytochrome C
- pro-apoptotic: Bax,Bak,Bad,Bid,Puma, etc.
- enhance release of cytochrome C
- DNA damages can trigger transcription factor
p53 to activate pro-apoptotic members
Apoptosis
Extrinsic and intrinsic pathways
- extrinsic pathways can also activate
intrinsic pathways to kill cells
- activate through pro-apoptotic Bid
- activated initiator caspase cleaves and
activate Bid to inhibit anti-apoptotic Bcl2
Apoptosis
- highly regulated process in cells
- can cause diseases if uncontrolled
- too much can cause Alzheimer’s,
Parkinson’s, ALS, etc.
- too little can cause cancers, autoimmune
diseases, etc.
- controlled by inhibitors of apoptosis
(IAPs) and extracellular survival factors
Apoptosis
Inhibitors of apoptosis (IAPs)
- internal cellular proteins that prevent
apoptosis
- bind and inhibit activated caspases
- some ubiquitylate caspases for degradation
- have to be inactivated by anti-IAP for
apoptosis to proceed
- IAP inhibitors include Smac/Diablo, etc.
Apoptosis
Survival factors
- external cellular signal proteins that
prevent apoptosis
- constant signalling between cells
required for survival
- control number of cells in tissues during
development to adulthood
- eg. required for neuronal survival
- “pruning” of neurons in development
- bind receptors to stimulate Bcl2 family
- activate Bcl2/ BclXL or inhibit Bad/Bax
 Apoptosis
Survival factors

Figure 18–12 Ways that extracellular survival factors can inhibit apoptosis
 Apoptosis
Survival factors
- eg: pruning in fetal development

Figure 18–11 Molecular Biology of the Cell

Apoptosis
Phagocytosis
- apoptotic cells are shrivelled to contain
degraded cellular contents
- do not trigger inflammatory response
- cell surface signals (eg.
phosphatidylserines) induce phagocytosis
- by macrophages
Apoptosis
Phagocytosis

Figure 23.3 Lippincott Cell & Molecular Biology

 Events of Apoptosis
APOPTOTIC SIGNALS

Chromatin condensation
DNA fragmentation

Phagocytosis
Nuclear breakup

Apoptotic cells &

Cell shrinkage, break
fragments
down into apoptotic
express cell
bodies
surface signals
 SUMMARY
Apoptosis
- normal process to balance cell growth
- remove damaged, excess, or “old” cells
- induced by cellular signals
- responsible for majority of cell death
- cells become shrivelled but are intact
- do not release cellular contents
- caspases and nucleases degrade cellular
contents
- removed by phagocytosis