APOPTOSIS

PY 1.3 and 1.4

 INTERCELLULAR JUNCTIONS
• In tissues, junction formed between the cells are called intercellular
junctions.
TYPES
 Tight junctions

Anchoring junctions

Gap junctions
 TIGHT JUNCTIONS
• Apical margins of epithelium of intestinal mucosa, renal tubular
epithelial cells, urinary tract, hepatobiliary tract and choroid plexus
• Also called as Zona occludens
• Located: apical region of cells
• Obliterates intercellular space near apical margin
• Made up of ridges. Form a barrier for transport of solutes and
solvents from lumen into interstitial space
• Membrane proteins: occludin, claudin, JAMS
 FUNCTION OF TIGHT JUNCTIONS
• Selective permeability barrier for macromolecules
• Presence of leaky channels: gut ---sodium ions pass freely while in
urinary bladder its nil
• Paracellular transport: depends on osmolality gradient
• Brain: tight junctions between endothelial cells of cerebral vessels
form effective blood brain barrier
• Ciliary bodies: tight junctions between inner non pigmented
epithelium to form blood aqueous barrier
 ANCHORING JUNCTIONS
• Cell to cell anchoring junctions
- Desmosomes
- Zonula Adherens

• Cell to Basal lamina anchoring junction

- Hemidesmosomes
- Focal Adhesions
 DESMOSOMES
• Focal thickening of two adjacent cell membranes

• Thickening: presence of dense layer of protein on cytoplasmic surface

• Separated by gap of 25nm

• Intermediary filaments from cytosol attached to thickened areas

• Intercellular space: cadherins

 ZONULA ADHERENS
• Located: below base of tight junctions

• Major site of attachment for microfilaments

• Cadherins are present in intercellular space

 HEMIDESMOSOMES
• HALF DESMOSOMES
• Microfilaments attached to it intracellularly
• Integrins are present

• FOCAL ADHESIONS
• Connect cell to basal lamina
• Actin filaments present intracellularly
• Assist in cell movement
 GAP JUNCTIONS
• Low resistance bridges which transport ions from one cell to another
• Intercellular space: 3nm
• Made up of: Connexons
• Each connexin has 6 identical protein subunits called Connexins which
surrounds an aqueous channel
• When connexon of adjacent cells are aligned continuous channel forms
which allows substances like ions to pass from one cell to another
• Connexons keep adjacent cell membranes at fixed gap so called Gap
junctions
 FUNCTIONS
 Electrical synapse: tissues behave like physiological syncytium
(cardiac muscle)

Also permits sugars and amino acids

Chemical messengers and hormones are also allowed

 APOPTOSIS (PROGRAMMED CELL DEATH)

• Greek word meaning falling off or dropping off

• Form of coordinated and internally planned cell death
• Genes of own cells play important role
• Examples
 Death of neurons in CNS during brain development and synapse
formation
 During fetal development degeneration of tissues like web of fingers
 Cells like eosinophils undergo apoptosis
MECHANISM OF APOPTOSIS
STIMULI
 Activation of cysteine proteases called caspases triggers apoptosis

Internal stimuli: Mitochondria releases a protein called smac -----

activates caspase 9 ----- induces apoptosis

External stimuli: Tumor necrosis factor activates caspase 8 ----

promotes DNA fragmentation and chromatin condensation
MOLECULAR MECHANISM OF
APOPTOSIS
• Initiation of apoptosis
 Triggered by absence of hormones, growth factors, cytokines

Activation of receptors like TNF receptors

Heat, radiation, hypoxia

Genetically programmed events

REGULATION OF APOPTOSIS
• Regulatory proteins are:
BCL-2: Located in outer mitochondrial membrane ,binds to BAX and
BAD proteins ----- promote apoptosis. May bind to BCL-XL protein that
inhibits apoptosis. Also binds to proapoptotic protease activating
factor (apzaf 1)

Others: caspases, p53 protein, hepatitis virus

STEPS OF APOPTOSIS
 PHYSIOLOGIC PROCESS
Examples
• Physiologic involution of cells – Endometrial shedding in menstrual
cycles, regression of lactating breast after cessation of breast feeding

• Normal shedding of intestinal epithelium

• Involution of thymus after childhood

 PATHOLOGIC PROCESS
• Tumour cell death on exposure to chemotherapeutic agents
• Transplant cell death by cytotoxic T cells that cause transplant
rejection
• Cell death by viral infections like viral hepatitis
• Prostatic atrophy following orchiectomy
• Cell death by radiation, hypoxia
• Degenerative diseases like Alzheimer’s disease, Parkinson’s disease
 CHANGES IN APOPTOSIS
• Cells become round, oval or reduce in size
• Cytoplasm becomes intensely eosinophilic with condensed or
fragmented nuclear chromatin
• Inflammatory response absent
• Apoptotic bodies formed
• Phagocytosis of apoptotic bodies
• Chromatic condensation around periphery of nucleus
 BIOCHEMICAL CHANGES
• Proteolysis of cytoskeletal proteins
• Cross linking of proteins
• Fragmentation of nuclear chromatin(nuclease)
• Thrombospondin, phosphatidylserine appear on outer surface of
apoptotic bodies which facilitates recognition by macrophages