APOPTOSIS

Willian Rengifo Apagueo

 APOPTOSIS

Definicion

La apoptosis es una muerte celular programada

dependiente de la energa para la eliminacin de
clulas individuales indeseadas.
 MUERTE CELULAR
Las clulas mueren por uno de dos
mecanismos: necrosis o apoptosis.
Dos procesos fisiolgicamente diferentes.
- Necrosis - muerte por lesin
- Apoptosis - muerte por suicidio
La apoptosis y la necrosis tienen diferentes
caractersticas.
 CARACTERISTICAS MORFOLOGICAS
Prdida de la integridad de la
membrana
Comienza con hinchazn de
citoplasma y mitocondrias.
Finaliza con lisis celular total,
sin formacin de vesculas,
lisis completa
Desintegracin (hinchazn) de
orgnulos.
Ampolla de membrana, pero
sin prdida de integridad
Comienza con la contraccin
del citoplasma y la
condensacin del ncleo
Finaliza con la fragmentacin
de la clula en cuerpos ms
pequeos
Las mitocondrias pierden
debido a la formacin de poros
que implican protenas de la
familia bcl-2.
 CARACTERISTICAS BIOQUIMICAS
Prdida de regulacin de la
homeostasis inica.
Sin necesidad de energia
La digestin aleatoria del
ADN (frotis de ADN despus
de la electroforesis en gel de
agarosa)
Fragmentacin de ADN
postltica (= evento tardo de
la muerte)
Proceso estrechamente regulado
Energa (ATP) -dependiente
La fragmentacin no aleatoria de
longitud mono- y
oligonucleosomal de ADN (tipo
escalera paterna)
Fragmentacin del ADN pretico
Liberacin de diversos factores en
el citoplasma por mitocondrias
Activacin de caspase en cascada
Alteraciones en la asimetra de la
membrana
 SIGNIFICADO FISIOLOGICO
Afecta a grupos de celdas
contiguas.
Evocado por alteraciones no
fisiolgicas (ataque del
complemento, virus lticos,
hipotermia, hipoxia, isquemia,
venenos metablicos)
Fagocitosis por macrfagos
Respuesta inflamatoria
significativa
Afecta celdas individuales
Inducido por estmulos
fisiolgicos (falta de factores
de crecimiento, cambios en el
ambiente hormonal)
Fagocitosis por clulas
adyacentes o macrfagos.
Ninguna respuesta
inflamatoria
 APOPTOSIS

Examples of apoptosis

Apoptosis in physiologic situations

Apoptosis in pathologic situations

 APOPTOSIS

Apoptosis in physiologic situations

In the human body about 100,000 cells are

produced every second by mitosis and a
similar number die by apoptosis*

* Vaux and Korsmeyer, 1999,Cell

Apoptosis in physiologic situations
Programmed cell death during embryogenesis

Formation of Development
free and Development of
independent of the brain reproductive
digits organs
Apoptosis in physiologic situations

Programmed cell death during adult stage

Death of cells
Cell loss in Elimination of
that have
proliferaing harmful self-
served their
cell reacttive
useful
populations lymhocytes
purpose
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.

Pathological
atrophy in
Accumulation Cell injury in
parenchymal
DNA damage of mis-folded certain
organs after
proteins infections
duct
obstruction
Morphology of Apoptosis
CELL SHRINKAGE

CHROMATIC CONDENSATION
 Biochemical features of Apoptosis

Protein By activation of caspases

Cleavage Caspases activate DNAses

DNA Cleavage into oligonucleosomes

By Ca2+-and Mg2+-dependent
Breakdown endonucleases

Phagocytic Phosphatidylserine
Recognition Thrombospondin
 Mechanisms of Apoptosis

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

Caspases are central initiators and executioners of apoptosis

Cysteine proteases
Caspase Cysteine-dependent
ASPartate-specific
s proteASES
 Mechanisms of Apoptosis

CASPASES

14 different members of the caspases-family have been described in mammals

Active cysteine residue in the catalytic site

Specificity in cleavage after an Asp residue

Synthesized as inactive zymogens (PROCASPASES)

 DNA FRAGMENTATION AND GEL
ELECTROPHORESIS
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 FRAGMENTATION - BIOCHEMICAL
HALLMARK OF APOPTOSIS
DNA cleaved into non-random fragments
180-200 bp fragments & multiples of this unit
Other morphological features of
apoptosis

Nuclear
breakdown
(Hoechst)
CELLS ARE BALANCED BETWEEN LIFE AND DEATH

DAMAGE Physiological death signals

DEATH SIGNAL

PROAPOPTOTIC ANTIAPOPTOTIC
PROTEINS PROTEINS
Two distinct pathways converge on
caspase acticvation

Mitochondrial pathway

Intrinsic pathway

The death receptor pathway

Extrinsic pathway
Mitochondria
contain several
proteins
that are capable of
inducing apoptosis

The choice between

cell survival and
death is determined
by the permeability
of mitochondria
The role of
mitochondria
in the
induction of
apoptosis
 Mitochondrial pathway
Intrinsic pathway
Mitochondria

BAX Cytochrome c release BCL-2

BAK BCL-XL
BOK BCL-W
BCL-Xs Pro-caspase 9 cleavage MCL1
BAD BFL1
BID DIVA
B IK NR-13
BIM Pro-execution caspase (3) cleavage Several
NIP3 viral
BNIP3 proteins

Caspase (3) cleavage of cellular proteins,

Nuclease activation,
Etc.

Death
 Extrinsic
pathway

The death
receptor
pathway
 Bcl-2 family members
A very large family with 30 members identified and belongs to both:

Bax BH1,
Bak BH2,BH3
Bok
Bcl -2
Bcl-XL Antiapoptotic
Bcl-W
Proapoptotic
A1
Mcl-1

Bid Noxa
BH1, Bim Puma BH3
BH2,BH3,BH4 Bik Blk
Bad BNIP3
Bmf Spike
Hrk
 APOPTOSIS

Physiological Intrinsic
receptor pathway damage pathway

MITOCHONDRIAL SIGNALS

Caspase cleavage cascade

Orderly cleavage of proteins and DNA

CROSSLINKING OF CELL CORPSES; ENGULFMENT

(no inflammation)
 APOPTOSIS

TOO MUCH: Tissue atrophy

Neurodegeneration
Thin skin

TOO LITTLE: Hyperplasia

Cancer
Athersclerosis
etc
 REFERENCES

Robins pathology
7th and 8th Edition

Introduction to apoptosis
By Andreas Gewies ApoReview in2003