APOPTOSIS

Many observations indicated cell death plays a considerable role during physiological processes

Programmed cell death

APOPTOSIS

Apoptosis is an energy dependent programmed cell death for removal of unwanted individual cells

 Cells die by one of two mechanisms Two physiologically different processes

Apoptosis and necrosis have different characteristics

 Loss of membrane integrity Begins with swelling of cytoplasm and mitochondria Ends with total cell lysis, no vesicle formation, complete lysis Disintegration (swelling) of organelles

Membrane blebbing, but no loss of integrity Begins with shrinking of cytoplasm and condensation of nucleus Ends with fragmentation of cell into smaller bodies Mitochondria become leaky due to pore formation involving proteins of the bcl-2 family.

 Loss of regulation of ion homeostasis No energy requirement Random digestion of DNA (smear of DNA after agarose gel electrophoresis) Postlytic DNA fragmentation (= late event of death)

Tightly regulated process Energy (ATP)-dependent Non-random mono- and oligonucleosomal length fragmentation of DNA(ladder type patern) Prelytic DNA fragmentation Release of various factors into cytoplasm by mitochondria Activation of caspase cascade Alterations in membrane asymmetry

 Affects groups of contiguous cells Evoked by non-physiological disturbances (complement attack, lytic viruses, hypothermia, hypoxia, ischemia, metabolic poisons) Phagocytosis by macrophages Significant inflammatory response

Affects individual cells Induced by physiological stimuli (lack of growth factors, changes in hormonal environment)

Phagocytosis by adjacent cells or macrophages No inflammatory response

APOPTOSIS

Apoptosis in physiologic situations Apoptosis in pathologic situations

APOPTOSIS

Apoptosis in physiologic situations

Vaux and Korsmeyer, 1999,Cell

Apoptosis in physiologic situations

Programmed cell death during embryogenesis

Formation of free and independent digits

Development of the brain

Development of reproductive organs

Apoptosis in physiologic situations

Programmed cell death during adult stage

Cell loss in proliferaing cell populations

Death of cells that have served their useful purpose

Elimination of harmful selfreacttive lymhocytes

Apoptosis in pathologic situations

Apoptosis eliminates cells that are genetically altered or injured beyond repair without eliciting a severe host reaction, thus keeping the damage as contained as possible.

DNA damage

Accumulation of mis-folded proteins

Cell injury in certain infections

Pathological atrophy in parenchymal organs after duct obstruction

Morphology of Apoptosis

Biochemical features of Apoptosis

Protein Cleavage DNA Breakdown Phagocytic Recognition

By activation of caspases Caspases activate DNAses

Cleavage into oligonucleosomes By Ca2+-and Mg2+-dependent endonucleases

Phosphatidylserine Thrombospondin

Mechanisms of Apoptosis
The fundamental events in apoptosis is the activation of enzymes called CASPASES

Caspases

Cysteine proteases Cysteine-dependent ASPartate-specific proteASES

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 ladderingof DNA

 DNA cleaved into non-random fragments 180-200 bp fragments & multiples of this unit

DAMAGE

Physiological death signals

DEATH SIGNAL

PROAPOPTOTIC PROTEINS

ANTIAPOPTOTIC PROTEINS

Mitochondrial pathway Intrinsic pathway The death receptor pathway Extrinsic pathway

Mitochondrial pathway Intrinsic pathway

Mitochondria

BAX BAK BOK BCL-Xs BAD BID B IK BIM NIP3 BNIP3

BCL-2 BCL-XL BCL-W MCL1 BFL1 DIVA NR-13 Several viral proteins

 Extrinsic pathway The death receptor pathway

Bcl-2 family members

A very large family with 30 members identified and belongs to both:
BH1, BH2,BH3

Antiapoptotic Proapoptotic

BH1, BH2,BH3,BH4

Bid Bim Bik Bad Bmf Hrk

Noxa Puma Blk BNIP3 Spike

BH3

APOPTOSIS

MITOCHONDRIAL SIGNALS

APOPTOSIS

REFERENCES

Robins pathology
7th and 8th Edition

Introduction to apoptosis
By Andreas Gewies ApoReview in2003