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Robbins: Pathologic Basis of Disease. 8th
Apoptosis
First described in 1972
From the Greek, meaning falling off cell death that: Occurs normally Eliminates unwanted or potentially harmful cells Cells that have outlived their usefulness Pathologic event Cells damaged beyond repair (especially to DNA)
(autoimmune) lymphocytes
lymphocytes
131 of 1090 somatic cells generated during development of this organism are eliminated by apoptosis. Caenorhabditis elegans CED-3: caspase homologue CED-4: Apaf-1 homologue CED-9: bcl-2 homologue
bodies
apoptotic bodies composed of cytoplasm and tightly packed organelles sometimes nuclear fragments NO INFLAMMATION
EM of apoptotic cell
EM of necrotic cell
Note Chromatin clumping Organelle swelling
Mediators of apoptosis
CASPASES
What are caspases?
Cysteine proteases that cleave after aspartic acid residue (Asp-X) At least 10 members have been identified Highly conserved across species
Mediators of apoptosis
CASPASES Involvement at two or more levels: initiator caspases: involved in decision/commitment to apoptosis (already discussed)
Caspase 8, 10 and 9
Caspase 3, 6 and 7
Mediators of apoptosis
CASPASES
Exist in pro-enzyme or zymogen form Undergo cleavage to be activated By other caspases or autocatalytically
Figure 1-24 p. 28
Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 2 December 2004 07:30 PM) 2004 Elsevier
superfamily
Contain cytoplasmic death domains that delivers the apoptotic signal TNF-R Fas
molecules become crosslinked by the Fas ligand Fas cytoplasmic death domains form a binding site for an adaptor protein (FADD) Also contains a death domain
and FADD binds inactive caspase-8 (pro-caspase-8) via death domains Multiple pro-caspase-8 molecules are brought into proximity and cleave one another to generate active caspase-8
cascade of caspase activation These other caspases mediate the execution phase of apoptosis This pathway can be inhibited by FLIP Binds to pro-caspase 8 but cannot cleave and activate it because it has no enzymatic activity Produced by viruses and some normal cells
Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 2 December 2004 07:30 PM) 2004 Elsevier
When Bcl-2/Bcl-x levels decrease, permeability of the mitochondrial membrane increases Causes leakage of proteins that can activate caspases Best known: cytochrome c
mitochondrial permeability And release of proapoptotic molecules into the cytosol No known role for death receptors
Once out in the cytosol, cytochrome c binds to apaf-1 (apoptosis-protease activating factor 1; ced-4 homologue) Complex of apaf-1 and cytochrome c activates caspase 9 Bcl-2 and Bcl-x may also directly inhibit apaf-1 activation Other mitochondrial proteins such as apoptosis inducing factor (AIF) can enter the cytosol Bind to and neutralize inhibitors of apoptosis Net effect: initiation of the caspase cascade
other signals stimulate the production of antiapoptotic member of the Bcl-2 family of proteins
pathway is the balance of pro- and anti-apoptotic molecules that Regulate mitochondrial permeability And release of death inducers that are normally sequestered in the mitochondria
Not well-defined May also be overlap between the extrinsic and intrinsic pathways (may not be distinct) Example: in hepatocytes Fas signalling activates proapoptotic bid which activates the mitochondrial pathway
Proteins involved in
transcription (c-Myc and NF-kB-inhibited) DNA replication DNA repair poly ADP-ribose polymerase(PARP) nuclear lamina (keratins 18, 19 and vimentin) elements of the cellular cytoskeleton (b-catenindisrupting cell-cell interactions, fodrin and gelsolin-disrupting the actin filament network)
Caspase 3
Activates DNA fragmentation factor 45 (DFF45) Activation of DFF45 in turn activates DFF40 which plays a critical role in the internucleosomal DNA degradation Also acts on mitochondrial substrates
disrupts electron transport loss of mitochondrial transmembrane potential
spacer regions first into large chunks (50300kb) then into multiples of 180 bp Form a ladder on agarose gels
markers on their surface that facilitate early recognition by nearby phagocytic cells
The loss of phospholipid asymmetry in the plasma membrane and translocation of phosphotidyl serine (PtdSer) to the outer leaflet of the lipid bilayer
molecules enhance uptake including Mannose binding lectins (MBL) and C1q
Limited inflammation
Uptake of apoptotic cells has been shown to
CTL-mediated lysis
Cytotoxic T cells recognize foreign antigens
presented by Class I MHC at the cell surface Upon recognition, CTL secrete
CTL-mediated lysis
Granzyme B: serine protease Cleaves proteins at aspartate residues Activates caspases By-passes up-stream signalling events Acts directly by inducing execution phase CTL also express Fas-L on their surface and
p53 mutation hormone-dependent (breast, prostate, ovary) Individuals with complement component C1q defects have an increased risk of developing systemic lupus erythematosus (SLE) and glomerulonephritis
autoimmunity
neurodegenerative diseases
ischemic injury