Unit 7: Apoptosis Notes

Unit 7: Apoptosis Notes

Introduction to Apoptosis

  • Apoptosis is a crucial process for maintaining tissue homeostasis.
  • It involves a balance between cell division and cell death.
  • The genetically regulated process of programmed cell death.
  • Most common form is apoptosis, preserved across unicellular organisms like yeast and bacteria.
  • Important for understanding processes in higher eukaryotes.

Importance of Apoptosis

  • Increased cell division or reduced apoptosis can lead to cancer progression.
  • Excessive cell death is linked to neurodegenerative diseases and autoimmune disorders.

Morphological Characteristics of Apoptosis

  • Cells undergoing apoptosis exhibit:
  • Shrinkage and condensation.
  • Collapse of cytoskeleton.
  • Fragmentation of chromatin and disassembly of the nuclear envelope.
  • Formation of apoptotic bodies due to membrane blebbing.
  • Contrast with necrosis, which is uncontrolled cell death that damages surrounding tissues and incites inflammation.

Phagocytosis in Apoptosis

  • Apoptotic cells are recognized and cleared by phagocytic cells (e.g., macrophages).
  • Phosphatidylserine exposure on the cell surface serves as a signal for recognition and clearance.
  • This prevents inflammation, highlighting a significant benefit of apoptosis.

Role of Apoptosis in Development and Immunology

  • Essential for developmental processes, e.g., tadpole tail removal in frogs and digit formation.
  • Crucial in the immune system for:
  • Maturation of B and T lymphocytes.
  • Removal of activated lymphocytes post-infection via apoptosis to prevent excess inflammation.

Mechanisms of Apoptosis: Caspases

  • Caspases are a family of cysteine proteases essential for apoptosis.
  • They are classified as initiator (e.g., caspase 8 and 9) and executioner caspases.
  • Caspase Activation: Zymogens (inactive precursors) are activated by apoptotic signals, resulting in a cascade effect.
  • Activated caspases cleave various substrates including nuclear lamins, cytoskeleton components, and cell adhesion proteins, leading to apoptotic changes.

Identification of Apoptotic Events

  • Various techniques to visualize apoptosis:
  • DNA laddering: Results from endonuclease activity, observed via gel electrophoresis.
  • TUNEL assay: Labels DNA ends generated during apoptosis (fluorescent visualization).
  • Annexin V staining: Binds to exposed phosphatidylserine on apoptotic cells, visualized by fluorescence or flow cytometry.
  • Mitochondrial membrane potential assays: Indicate loss of membrane potential and release of cytochrome c.

Apoptotic Pathways

  • Two main pathways for apoptosis: extrinsic and intrinsic.
Extrinsic Pathway
  • Initiated by binding of ligands to death receptors on the cell surface (e.g., Fas ligand).
  • Triggers an intracellular cascade through Death Inducing Signaling Complexes.
  • Certain proteins can inhibit this pathway by mimicking receptors without the death domain.
Intrinsic Pathway
  • Activated by internal stimuli (e.g., DNA damage, hypoxia).
  • Relies on mitochondrial function; release of cytochrome c into the cytoplasm is crucial.
  • Cytochrome c binds to Apaf-1, leading to apoptosome formation and initiation of downstream caspase activation.

Regulation of Apoptosis: BCL-2 Family

  • BCL-2 proteins play a key role in regulating the intrinsic pathway.
  • Anti-apoptotic: BCL-2 and BCL-XL inhibit apoptosis by preventing cytochrome c release.
  • Pro-apoptotic: BAX and BAK promote apoptosis by forming pores in the mitochondrial membrane, allowing cytochrome c to escape.
  • The balance between these opposing forces regulates cell survival or death.

Conclusion

  • Understanding the mechanisms and pathways of apoptosis is crucial for insights into cancer and other diseases.
  • Research continues to explore these pathways for therapeutic interventions in diseases characterized by abnormal cell death or survival.