APOPTOSIS DEFINITION
❖Genetically programmed death of a single cell (suicide of cell).
❖Organism used for most of apoptotic studies:
Caenorhabditis elegans (Nematode).
❖Term 'Apoptosis' means 'Falling off'.
MECHANISM OF APOPTOSIS
Apoptosis results from the activation of enzymes called
caspases (so named because they are proteases
containing a cysteine in their active site and cleave
proteins after aspartic residues).
The Mitochondrial (Intrinsic) Pathway of Apoptosis
The mitochondrial pathway is responsible for apoptosis in
most physiologic and pathologic situations.
KEY REGULATORS AND ENZYMES
Pro-apoptotic regulators: BAX, BAC.
Anti-apoptotic regulators: BCL family (BCL-2, BCL-XL, MCL-I).
Stress Sensors : BIM, BID, BAD, PUMA, NOXA
Enzymes:
1. Caspases:
❖ Contains cysteine.
❖ Cleaves near aspartic acid residue.
❖ Types:
• Initiator caspases: 8, 9, 10.
• Executioner caspases: 3, 6, 7.
2. Endonuclease:
❖ Breaks down DNA to fragments.
Anti-apoptotic : BCL2, BCL-XL, and MCL1 are the principal members
of this group; they possess four BH domains (called BH1-4). These
proteins reside in the outer mitochondrial membrane as well as in the
cytosol and ER membranes.
Pro-apoptotic : BAX and BAK are the two prototypic members of this
group; they contain the first three BH domains (BH1-3). On activation,
BAX and/or BAK oligomerize within the outer mitochondrial
membrane and enhance its permeability. The precise mechanism by
which BAX-BAK oligomers permeabilize membranes is not settled.
Regulated apoptosis initiators : Members of this group, including
BAD, BIM, BID, Puma, and Noxa, contain only one BH domain, the third
of the four BH domains, and hence are sometimes called BH3-only
proteins.
The Extrinsic (Death Receptor–Initiated) Pathway of Apoptosis
This pathway is initiated by engagement of plasma membrane death receptors.
Death receptors are members of the tumor necrosis factor (TNF) receptor family
that contain a cytoplasmic domain involved in protein-protein interactions.
This death domain is essential for delivering apoptotic signals.
The Execution Phase of Apoptosis
The intrinsic and extrinsic pathways converge to activate a caspase cascade that
mediates the final phase of apoptosis.
MORPHOLOGY
The following morphologic features, some best seen with the electron microscope, characterize cells
undergoing apoptosis.
Cell shrinkage : The cell is smaller in size, the cytoplasm is dense (Fig. 2-22A), and the organelles,
although relatively normal, are more tightly packed. (Recall that in other forms of cell injury, an
early feature is cell swelling, not shrinkage.)
Chromatin condensation : This is the most characteristic feature of apoptosis. The chromatin
aggregates peripherally, under the nuclear membrane, into dense masses of various shapes and
sizes (Fig. 2-22B). The nucleus itself may break up, producing two or more fragments.
Formation of cytoplasmic blebs and apoptotic bodies : The apoptotic cell first shows extensive
surface blebbing, then undergoes fragmentation into membrane-bound apoptotic bodies composed
of cytoplasm and tightly packed organelles, with or without nuclear fragments (Fig. 2-22C).
Phagocytosis of apoptotic cells or cell bodies, usually by macrophages : The apoptotic bodies
are rapidly ingested by phagocytes and degraded by the phagocyte's lysosomal enzymes.
Plasma membranes are thought to remain intact during apoptosis, until the last stages, when they
become permeable to normally retained solutes.
On histologic examination, in tissues stained with hematoxylin and eosin, the apoptotic cell appears
as a round or oval mass of intensely eosinophilic cytoplasm with fragments of dense nuclear
chromatin. Because the cell shrinkage and formation of apoptotic bodies are rapid and the pieces
are quickly phagocytosed, considerable apoptosis may occur in tissues before it becomes apparent
in histologic sections.
In tissues, apoptosis—in contrast to necrosis—does not elicit inflammation, making it more difficult
to detect histologically.
Morphologic features of aptosis
Necrosis vs Aptosis
Feature Necrosis Apoptosis
Enzymatic/Ischemic Genetically programmed cell
Definition process death
Type of process Passive process Active process
Cells involved Group of cells Single cell
Always pathological Physiological/pathological
Occurrence Accidental Suicidal
Cell size Increases Decreases
Cell membrane Affected Intact
Inflammation Present Absent
Markers Absent Annexin V, CD 95
Step ladder pattern due to
Polyacrylamide gel electrophoresis Smear pattern (Absent
endonuclease activity; breaks
(PAGE)/DNA electrophoresis endonuclease)
DNA to fragments
SUMMARY
❖ Apoptosis is a crucial process for maintaining cellular health.
❖ Proper regulation prevents disorders like cancer or autoimmune diseases.
❖ Studying apoptosis helps understand cell biology and therapeutic targets.
References
❖ Kumar, V., Abbas, A. K., & Aster, J. C. (2021). Robbins & Cotran Pathologic Basis
of Disease (10th ed., South Asia Edition, Vol. 1). Elsevier.
