Cell Death

Cell Death
• The body is very good at maintaining a constant
number of cells. So there has to exist
mechanisms for ensuring other cells in the body
are removed.
• Two forms of cell death
– Apoptosis - suicide - Programmed Cell Death
– Necrosis - killing - decay and destruction
Etiology of cell death
Major Factors

Accidental Genetic

Necrosis Apoptosis
Necrosis: consequences of cell injury
Contrast ofApoptosis and necrosis

Apoptosis Necrosis
Death by apoptosis is a neat, orderly process
A、Normal cell B、Apoptosis: Apoptotic bodies
 Apoptosis: a physiological response to
Necrosis: a pathological response
specific suicide signals, or lack of
to cellular injury
survival signals

Chromatin condenses and migrates to nuclear

membrane. Internucleosomal cleavage leads
Chromatin clumps
to laddering of DNA at the nucleosomal repeat
length, ca. 200 bp.
Mitochondria swell and rupture Cytoplasm shrinks without membrane rupture
Plasma membrane lyses Blebbing of plasma and nuclear membranes

Cell contents are packaged in membrane

Cell contents spill out bounded bodies, internal organelles still
functioning, to be engulfed by neighbours.
Epitopes appear on plasma membrane
General inflammatory response is
marking cell as a phagocytic target.
triggered
No spillage, no inflammation
www.chembio.uoguelph.ca
 Apoptosis Necrosis
Physiological or pathological Always pathological
Single cells Sheets of cells
Energy dependent Energy independent
Cell shrinkage Cell swelling
Membrane integrity maintained Membrane integrity lost
Role for mitochondria and
No role for mitochondria
cytochrome C
No leak of lysosomal enzymes Leak of lysosomal enzymes
Characteristic nuclear changes Nuclei lost
Apoptotic bodies form Do not form
DNA cleavage No DNA cleavage
Activation of specific proteases No activation
Regulatable process Not regulated
Evolutionarily conserved Not conserved
Dead cells ingested by neutrophils
Dead cells cause inflammation
and macrophages
2. Biological functions of
apoptosis
 Control of cell numbers and cell size
• Three processes operate to control the eventual form
a body part takes
– Cell growth (survival factors, growth factors)
– Cell division (mitogenic factors)
– Cell death (apoptotic factors)
• Single celled organisms grow as fast as they are able
to limited by factors such a food availability
• Multicellular organisms receive signals from other
cells in the body
 APOPTOSIS AS A PHYSIOLOGICALLY
IMPORTANT PROCESS

In embryonic • Tissue developmental programs which

and fetal control sculpting of embryonic form
development • Developmental organization of the
nervous system
• Elimination of self-reactive components
of the immune system
In the adult • On stimulation by T-lymphocytes
• In response to DNA damage or
abnormality, e.g. by radiation, viral
infection or transformation
• In certain organs and tissues, on
withdrawal of supporting hormones

www.chembio.uoguelph.ca
 Functions of apoptosis
Sculpt body structures, e.g. hand digit
 Functions of apoptosis

Serve some function but no longer needed

e.g. tadpole tail of frog.
 Functions of apoptosis

Needed in one sex but not another e.g. Mullerian duct important
for female is eliminated in males by apoptosis.
 Functions of apoptosis

Serve in immune system as a defense

mechanism to get rid of harmful or
damaged cells.
 Produced in excess, e.g. extra neurons are
removed by apoptosis during neurogenesis.
3.Molecular mechanisms
of apoptosis
 1090 –131= 959(cells)

PRIZED Horvitz, and Sulston share Physiology or Medicine Nobel

（2002）

“for their discoveries concerning genetic regulation

of organ development and programmed cell death”
 Early researches (MIT: Robert Horrid , 1986)
C. elegans:1090 cells, 131 cells death.

The Finding of CED3 mutant

Without losing any of their
cells to apoptosis.
C elegans: a millimeter long,
CED3 gene play a crucial role transparent body only a few cell
in the process of apoptosis. types, from zygote to mature
adult only in 3.5days.
 APOPTOSIS in C.elegans

C.elegans genome: 19099 genes (790 seven-pass

transmembrane receptors, 480 zinc finger proteins, and 410
protein kinases)
The life cycle of C. elegans from egg to sexual maturity (and
new eggs) is about 3 days
ced-1, -3, -4, and -9 (Cell death determining) proteins in
C.elegans are closely related to mammalian apoptosis-
1090 –131= 959(cells) regulating genes

The adult hermaphrodite consists of exactly 959 somatic

cells of precisely determined lineage and function.
Individual cells are named and their relationships to their
neighbors are known

Overall, the 959 somatic cells of adult C.elegans arise

from 1090 original cells; exactly 131 somatic cells undergo
programmed cell death in the wild type worm

Of the 1090 cells, 302 are neurons, and many of the

programmed deaths also lie in the neuronal lineage
www.chembio.uoguelph.ca
 Morphological and biochemical
characteristics of apoptosis
 Morphologi changes:
Early : Chromosome condensation, cell body shrink
Later : Blebbing and Nucleus and cytoplasm
fragment—Apoptotic bodies
At last: Phagocytosed
Concanavalin A (green) stains outlines of the
apoptotic surface blebs

red - propidium- jodide

apoptotic human keratinocyte

What "signals" induce apoptosis?

Signals can be extracelluar:

• a hormone (such as thyroxine which causes apoptosis
in tadpole tails
• a lack of a "survival" signal (which inhibits apoptosis)
such as a growth factor
• a cell:cell contact from an adjacent cell

Signals can be intracelluar:

• ionizing radiation
• virus infection
• oxidative damage from free radicals
 Mechanism of Apoptosis
• Internal signals: Mitochondrial pathway.

• External signals: Death receptor pathway.

• Apoptosis-Inducing Factor.
Internal signals: Mitochondrial
pathway
External signals: Death receptor
pathway
 Apoptosis-Inducing Factor (AIF)
• Release from the mitochondria.
• Migrates into the nucleus.
• Bind to DNA.
• Destruction of the DNA
• Cell death.
(1) Extrinsic pathway: Fas Signaling Pathway

Receptor-mediated pathway of apoptosis

Fas (also called Apo-1 or CD95) is a member of the

tumor necrosis factor receptor (TNFR) superfamily.
(2) Intrinsic pathway: Mitochondrial pathway

Various types of internal stimuli

(DNA damage, high Ca2+ , Oxidative stress)

Bcl-2 family: Bad or Bax to become inserted into outer

membrane of mitochondria

Release of cyt-c from mitochondria

Apoptosome (cyt-c, Apaf-1, procaspase 9 complex) →

Caspase Cascade
 Sinyal apoptosis
Fas ligan

Reseptor Fas

Bid Caspase-8
FADD
mitokondria
Procaspase-8
Bcl-2

sitokrom-c
AIF Apaf-1 Procaspase-9 Smac

IAP
Caspase-9

Caspase-3

Caspase cascade

CAD

DNA fragmentasi aktivasi

(apoptosis) penghambatan
 Summary of death receptor pathway
ligands

Death receptors

FADD/DISC

Caspase 8, 10

Caspase 3, 6, 7
procaspase 3 caspase 3

Lamin, CAD,actin,
Adhesion molecules, etc
 Bcl-2 family proteins control cytochrome c
translocation

• Bax forms homo-dimers in the presence of apoptotic signals.

This homodimer presumably promotes the opening of a
channel that controls the translocation of cytochrome c
from the intermembrane space to cytoplasm

cytoplasm

intermembrane
space

• Bcl-2 interferes with Bax function by forming a hetero

dimer with Bax. This leads to the closing of the channel and
inhibition of cytochrome c translocation
 Cytochrome c translocation initiates caspase cascade
Apaf-1
• In the cytosol, cytochrome c binds to
Apaf-1 to form apoptosome

• The apoptosome recruits procaspase 9

to generate the active caspase 9 Cyt. c

• Caspase 9 then activates the

executioner caspase 3, 6, 7

Procaspase 9
Caspase 9
Apoptosome

Procaspase 3

Caspase 3
 Extrinsic and intrinsic pathways converge at
executioner caspases
ligands Intrinsic apoptosis signals

Death receptors Pro-apoptotic Bcl-2 proteins

Cytochrome c translocation
FADD/DISC

Apoptosome (Apaf-1/cyt.c)
Caspase 8, 10
Caspase 9

Caspase 3, 6, 7

Lamin, CAD,actin,
Adhesion molecules, etc