APOPTOSIS
APOPTOSIS
Apoptosis is defined as the natural or programmed cell death that
occurs under genetic control.
• When cells are no longer needed or become a threat to the
organism, they undergo PCD or apoptosis.
• Apoptosis is a Greek word meaning “falling off”.
• Since the cell’s own genes play an active role in its demise,
apoptosis can also be called as cell suicide.
Apoptosis is a pathway of cell death that is induced by a tightly
regulated suicide program in which cells activate enzymes that
degrade the cells own nuclear DNA and nuclear and cytoplasmic
proteins.
HISTORY
German scientist Carl Vogt put forth the principle of apoptosis in
1842.
Walther Flemming an anatomist described the process of
programmed cell death in 1885.
John Kerr 1965, while studying tissues using electron
microscopy, was able to distinguish apoptosis from traumatic
cell death.
CELL DEATH
Cell die by one of the two mechanism-
1. Necrosis-Death by injury
2. Apoptosis-Death by suicide
APOPTOSIS v/s NECROSIS
Cell death occurs in two ways : necrosis and apoptosis.
Apoptosis should be distinguished from necrosis in which the
healthy cells are destroyed by some injury inflicted on the cells.
Since necrosis is caused by some external process, it can also
be called as cell murder.
Need of Apoptosis
The purpose of apoptosis is to remove unwanted cells without
causing any stress or damage to the neighboring cells.
It plays a role in cellular homeostasis.
Useful in removal of cell that is damaged beyond repair by toxins
or viruses.
It is an essential event during developmental stages and in
adults. Therefore, decreased apoptosis has been implicated in
the genesis of malignancy and autoimmune diseases.
Reasons for Apoptosis
Withdrawal of positive signals
Growth factors for neurons
IL – 2
Receipt of negative signals
Increased levels of oxidants within the cell
Damage to DNA by oxidants
Death activators
TNF α
TNF β
Fas ligand
CASPASES
Apoptosis results from the activation of enzymes called CASPASES
Cysteine-dependent ASPartyl-specific proteASE(CASPASE)
INITIATOR CASPASES-
Caspase 2
Caspase 8
Caspase 9
Caspase 10
EXECUTIONER CASPASE
Caspase 3
Caspase 6
Caspase 7
optosis may be divided into two phase
nitiation phase
Activation of initiator caspases
Two distinct but convergent pathways
The Intrinsic(Mitochondrial)Pathway
The Extrinsic(Death Receptor-Initiated) Pathway
xecution Phase
Activated initiator caspases activate executioner caspase to cause cell death
Mechanism of Apoptosis
1.Anti-apoptotic
proteins(Prevent apoptosis) Prevent leakage of
cytochrome c into cytosol by keeping the mitochondrial membrane
impermeable Prevent apoptosis
BCL2,BCL-XL and MCL1
Present in outer memebrane as well as cytosol and ER membranes
2.Pro-apoptotic proteins(Induce apoptosis) Form a channel in the outer
mitochondrial membrane Leakage of cytochrome c in to cytoplasm
Activation of caspase Induced apoptosis
BAX and BAK
3.Sensors(Arbiters of apoptosis) Sometimes called BH3-only
proteins.Sense the cellular stress ans damage.Regulate the balance
between the other anti and pro-apoptotic proteins.
BAD,BIM,BID,Puma and Noxa
• Apoptosis depends upon the balance between the pro and anti
apoptotic proteins
• In a healthy cell, anti-apoptotic proteins binds with pro-apoptotic
proteins thereby blocking their action.
• If a cell is damaged or it stops receiving survival signals, Bcl – 2
and Bcl –x are blocked in turn.
• Bax and Bac are then free to punch a series of channels in the
mitochondria.
INTRINSIC PATHWAY
Initiated from within the cell.
Activated in response to signals such as DNA damage, loss of
cell survival factors ,cell stress.
Hinges on balance brtween pro and antiapoptotic signals of Bcl-2
family.
Apaf-1,cytochrome-c,ATP(apoptosome) activate procaspase-9
complex.
pro apoptotic proteins released which activate caspase
proteases
EXTRINSIC PATHWAY
Begins outside of the cells.
Activation of death receptors (Fas-R,TNF-R ,DR- 3,DRY/DR5) by
death ligands (Fas-L,TNF- alpha,Apo3L,Apo2L)play major role.
Death induced signalling compex (DISC) activated.
On DISC activation same effctor pathway as intrinsic pathway is
adopted
Pathophysiological changes in Apoptosis
Apoptotic cells become round or oval and reduce in size.
Cytoplasm becomes intensely eosinophilic.Though cytoplasm is
reduced, organelles remain almost normal.
Chromatic condensation occurs.
Nuclear fragmentation occurs.
Apoptotic bodies contain compacted organelles.
Phagocytosis of apoptotic bodies occur by macrophages. The
phagocytic cells secrete cytokines like IL-10 and TGF-β which
inhibit inflammation of neighbouring tissues.
Biochemical changes
Proteolysis of cytoskeletal proteins.
Cross – linking of protein molecules.
Fragmentation of nuclear chromatin by activation of nuclease.
A glycoprotein – thrombospondin and a phosphoprotein –
phosphatidylserine appear on the outer surface of apoptotic
bodies for recognition by macrophages.
Physiological processes due to Apoptosis
The separation of fingers and toes in a developing human
embryo occurs because cells between the digits undergo
apoptosis.
Endometrial shedding in menstrual cycles.
Regression of lactating breast after cessation of breastfeeding.
Normal shedding of intestinal epithelium.
Involution of thymus after childhood.
Pathological processes as a result of Apoptosis
Tumour cell death on exposure to chemotherapeutic agents.
Transplant cell death by cytotoxic T cells that cause transplant
rejection.
Cell death induced by viral infections.
Cell death induced by radiation, hypoxia.
Degenerative diseases like Alzheimer’s disease, Parkinson’s
disease etc.
Detection of Apoptosis
DNA fragmentation assay by electrophoresis.
TUNEL (terminal deoxynucleotidyl transferase dUTP nick end
labeling) staining.
Demonstration of chromatic condensation by H & E staining.
Estimation of cytosolic cytochrome – C, activated caspase
