APOPTOSIS

Dr. MARUTHI DEVI

PG STUDENT
ORALPATHOLOGY
16-07-2007

CONTENTS

INTRODUCTION
DEFINITION
CLASIFICATION
MECHANISM
MORPHOLOGIC CHANGES
MOLECULAR CHANGES
BIOCHEMICAL CHANGES
HISTOPATHOGENESIS
THE SIGNALS INDUCING APOPTOSIS
TECHNIQUES TO IDENTIFY APOPTOTIC CELLS
CONCLUSION

INTRODUCTION

In 1971,John Kerr,an Australian pathologist,

described cell death in rat liver after ligation of a
branch of the portal vein as shrinkage necrosis.
Kerr&co-workers further described the morphology
of this form of cell death & used the term Apoptosis .

DEFINITION
Apoptosis is a genetically regulated
evolutionary conserved process that removes
damaged cells from a tissue or organ.

It is a form of co-ordinated and internally

programmed cell death. Hence living cells
posses inherent inhibitory mechanism to
keep apoptotic process in check.

CLASSIFICATION
1.
2.

Physiological apoptosis
Pathological apoptosis


1.

2.
3.
4.
5.
6.

PHYSIOLOGICAL APOPTOSIS
The programmed destruction of cells during
embryogenesis
Hormone dependent involution in the adult
Cell deletion in proliferating cell population
Death of host cells
Cell death induced by cytotoxic Tcells
Involution of Thymus in early life

PATHOLOGICAL APOPTOSIS
1.

Cell death by injurious stimuli

Radiation & cytotoxic anti cancer drugs

Heat & Hypoxia
Accumulations
Viral diseases
Pathological atrophy in parenchymal organ
Cell death in tumors
Cell death by cytotoxic T-cells in immune mechanism
Progressive depletion of CD4 &T cells in AIDS
In degenerative diseases of C.N.S

2.
3.
4.
5.
6.
7.

MORPHOLOGICAL CHANGES
Death

of a single cell
No inflammatory reaction
Cell shrinkage
Cyotoplasmic blebs on membrane
Apoptotic bodies
Chromatin condensation
Phagocytosis of apoptotic bodies by
macrophages

.CELL SHOWING APOPTOSIS

BIOCHEMICAL CHANGE
Proteolysis

of cytoskeletal proteins
Protein-protein cross linking
Fragmentation of nuclear chromatin by
activation of nuclease
Appearance of phosphatidylserine on the
outer surface of cell membrane
Appearance of an adhesive glycoprotein
Thrombospondein on Apoptotic bodies

MECHANISM OF APOPTOSIS
1.

INITIATION

EXTRINSIC PATHWAY OR RECEPTOR

INITIATED
INTRINSIC PATHWAY OR MITOCHONDRIAL
PATHWAY
EXECUTION PHASEP
PHAGOCYTOSIS

2.
3.

INITIATION PHASE
EXTRINSIC

PATHWAY

INTRINSIC PATHWAY

THE EXECUTION PHASE

A.

Execution caspases act & cleave

cytoskeleton & nuclear matrix proteins. This
disrupt the cytoskeleton& lead to break down
of nucleus & DNA damage.

PHAGOCYTOSIS
Apoptotic

cells & their fragments have marker

molecules on their surfaces which facilitates
early recognition by adjacent cells or
phagocytes for phagocytosis.
Phosphatidylserine which is present in he
inner membrane of the cell come to lie on the
outer surface of the cell membrane as the cell
undergoes apoptosis & acts as a marker.

HISTOPATHOLOGICAL CHANGES
The characteristic changes by light microscopy,
1.condensation of the nuclear chromatin-pyknosis
2.cell shrinkage & loss of cell-cell contact.
Increasing eosinophilia of the cytoplasm.
3.the cytoplasmic organelles are still preserved at this stage .
4.as the process continues the nuclear chromatin brakes up into
fragments-karyohexis
5.Dissolution of nuclear membrane.
6.cytoplasmic blebs break away from the cell surface & entire cell
breaks up to form membrane bound fragments.
7.cell fragments containing nuclear material are known as
apoptotic bodies

THE SIGNALS THAT INDUSE APOPTOSIS

Apoptosis

after growth factor deprivation

DNA damage mediated apoptosis.
Apoptosis induced by tumor necrosis factor
family receptors.
Cytotoxic T-lymphocyte mediated apoptosis.

TECHNIQUES TO IDENTIFY APOPTOTIC CELLS

Haematoxylin ,Feulgen, or acridine orange

For staining chromatin condensation.
Flow cytometry to visualize rapid cell shrinkage.
In situ technique or gel electrophoresis to detect
DNA changes.
Annexin-v as marker for apoptotic cell membrane
having phosphatidylserine on the cell exterior.
Agrose gel analysis, PCR analysis, COMET assays,
ELISA system, TUNEL,to identify fragments.

CONCLUSION

Our understanding of Apoptosis and

mechanism of normal development and
pathogenesis of many diseases including
AIDS ,Neurodegenarative diseases, and Cancer
and the role of Apoptosis in these diseases has
lead to the development of new and more
specific therapies.

REFERENCES
ROBINS

&COTRAN -7th EDITION

HARSHMOHAN

-4th EDITION

JOURNAL

OF ORAL PATHOLOGY &

MEDICINE, vol-32,no-3, March 2003

JOURNAL

OF ORAL PATHOLOGY &

MEDICINE, vol-31,no-3,March 2002