24. Apoptosis And Necrosis Flashcards
Increased in muscle size
Hypertrophy
Can act as a way for muscle cell to adapt to increase workload.
Two types of cellular response to stress and injury
Physiological
Pathological
Adaptation to a stimulus within the normal range
Physiological Response
Adaptation to a stimulus outside the normal physiological range
May result in cellular malfunction, damage, or death
Pathological Response
Fate of the cell exposed to a stimulus depends on:
Type of stimulus
Duration
Magnitude
Vulnerability of cell
Common causes of Cell injury
Metabolic
Oxygen deprivation
Hypoxia: oxygen deficiency
Ischemia: Blood flow deficiency
Nutritional
Deficiency, excess
Common causes of Cell injury
Chemical
Drugs, alcohol, poisons
Common causes of Cell injury
Physical
Trauma, extreme temperatures, radiation
Common causes of Cell injury
Biological
Virus, bacteria, parasite
Common causes of Cell injury
Immunological
Allergic reactions, autoimmune disease
Common causes of Cell injury
Genetic
Chromosomal abnormalities
Mutations
Common causes of Cell injury
Aging
Cellular senescence
Common causes of cell injury
Four major intracellular systems affected
- Membrane integrity (cellular plasma membrane, organelle membranes)
- ATP production
- Protein synthesis
- Genomic or chromosomal integrity
Early cellular responses to injury
Earliest evidence to injury
Loss of normal staining intensity due to swelling of organelles
Cloudy swelling
Rough Endoplasmic Reticulum becomes dilated with the loss of surface ribosomes.
Loss of cytoplasmic free ribosomes.
Early Cellular Responses to injury
Continued swelling of organelles
Vacuoles appear in cytoplasm (stains faintly with loss of basophilia)
Hydropic degeneration
Early cellular response to injury
Accumulation of triglycerides in cytoplasm
Most common in liver (rarely in skeletal muscle
Common causes (toxins, alcohol, chronic hypoxia)
Fatty change (steatosis)
Cell accumulation disorders (CLeG V)
CLeG V
Cancer
Lupus erythematosus
Glomerulonephritis
Viral infections
Cell Loss disorders (MAAAP)
MAAAP
Myocardial infarction
Alzheimers
AIDS
Aplastic anemia
Parkinson’s
Cell death
Cell swelling, damage to plasma membrane, random DNA degradation
Necrosis
Principle outcome in many injuries (ex. Ishemia, toxins, infections & trauma)
Cell death
Cell shrinkage, plasma membrane blebbing, aggregation of chromatin, fragmentation of nucleus, oligonucleosomal DNA fragmentation, caspase cascade activation
Apoptosis
Regulated cell suicide program
Occurs during development and throughout adulthood
Physiological (development, tissue homeostasis)
Some pathological (DNA damage, misfolded proteins, viral infections)
Pathological, acute cell injury, cell unable to maintain homeostasis, cell swelling, loss of plasma membrane integrity, cell contents released, surrounding tissue damage, inflammation
Necrosis
Physiological (planned), genetic, programmed cell death, cell shrinking, DNA aggregation, maintains plasma membrane integrity, no surrounding tissue damage, no inflammation
Apoptosis
Assessment of Necrosis
Morphilogical
Live cell imaging
Intercalates & labels DNA
If positive=leaky/discontinuous plasma membrane =necrosis
Propidium Iodide (PI) staining
Assessment of Necrosis
Morphilogical
Increased eosinophilia (loss of cytoplasmic RNA, increase in denatured proteins)
Variable nuclear staining (typical loss of basophilia & total loss of nuclear staining after a couple days; sometimes pyknosis visible)
H&E staining
Assessment of Necrosis
Morphilogical
Discontinuous plasma & organelle membranes
TEM
Assessment of Necrosis
Biochemical
Random DNA degradation
Increased levels of lactate dehydrogenase
Necrosis of the kidney
Apoptosis
Steps of apoptosis
Intracellular
- Phosphotidylserine moves from inner to outer leaflet
- Membranes bleb
- Condensed chromatin is found in nucleus.
- Cytochrome C is released from the mitochondria
Steps of apoptosis
Extracellular
- Phosphotidylserine is exposed on the surface of apoptotic cells
- The apoptotic cell is engulfed or phagocytosed by a macrophage
- The engulfed apoptotic cells is internalized then degraded within the macrophage
- The macrophage releases cytokines IL-10 and TGF-beta to inhibit inflammation.
Webbing of human toes may occur due to…
…interruption of apoptosis
Abnormal tissue homeostasis
Loss of cells due to apoptosis
Immune deficiency syndromes, some types of anemia
Abnormal tissue homeostasis
Irreplaceable due to non-renewing cell populations (brain or heart)
Alzheimer’s, Parkinsons, myocardial infarction
Abnormal tissue homeostasis
Accumulation of cells due to failure of apoptosis
Malignant neoplasia, autoimmune syndromes
Two mechanisms of apoptosis
Intrinsic (mitochondrial)
Extrinsic (Death receptor)
Differ in induction and regulation
Both result in activation of initiator & executioner caspases
Intrinsic B C A 9 C 3
Death signal (DNA damage) —> BAX (pro-apoptotic protein upregulated) —> Release of Cytochrome C from Mitochondria —> Activates Apaf-1 (assembly of Apoptosome) —> Activates Procaspase 9 (initiator) —> Activation of the Caspase cascade —> Activates Caspase 3 (activator).
BCA9C3
Extrinsic F FT D C83
Fas ligand (binding of ligand to death receptor) —> FADD & TRADD (recruitment of death adaptor proteins) —> DISC formed (death inducing signaling complex) —> Caspase Cascade —> Caspase 8 (initiator) —> Caspase 3 (effector caspase).
Bcl-2 Protein Family
Two functional classes
Pro-Apoptotic
Anti-Apoptotic
Bcl-2 protein family
Pro-apoptotic
Bid (Links extrinsic & intrinsic pathways
Bax & Bak (permeabilization of mitochondrial outer membrane)
Bcl-2 Protein Family
Anti-apoptotic
Bcl-2
Bcl-xL
Assessment of Apoptosis
Caspase Activity
Assessment of Apoptosis
Annexin 5
Protein binds to phosphotidylserine when exposed on the outer leaflet of plasma membrane.
Assessment of Apoptosis
DNA Laddering
DNAse cleaves intermucleosomal DNA
180-200 bp fragments
Assessment of Apoptosis
TUNEL
Terminal Uridine deoxyNucleotidyl transferase nick End Labeling
Detects DNA fragmentation
DNA Laddering
Agrose gel electrophoresis of DNA from cultured cells
A. Viable cells
B. Apoptotic Cells
C. Necrotic Cells
TUNEL
Assay detects DNA fragmentation
Tagged or labeled deoxynucleotides to 3’ DNA ends
TUNEL
Assay detects DNA fragmentation
Tagged or labeled deoxynucleotides to 3’ DNA ends