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Souvik Chatterjee
INDEX
Introduction Definitions Classification Tight junctions Adherens junctions Desmosomes Gap junctions Hemidesmosomes and focal contacts Cellular adhesion Techniques for diagnosis of intercellular junctions Summary
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INTERCELLULAR JUNCTIONS
It is a type of structure that exists within the tissue of
epithelia, nerves and muscles depends on the ability of the cells to interact with each other and the extracellular matrix. Plasma membrane specializations called cellular junctions ,mediate this interaction.
Intercellular junctions are specialized regions on the
Contd
Cell junctions consist of protein complexes and provide contact
between neighbouring cells or between a cell and the extracellular matrix. They are also called membrane junctions
They also build up the paracellular barrier of epithelia and control
the paracellular transport. Cell junctions are especially abundant in epithelial tissues.
On the molecular level intercellular junctions consists of three
OCCLUDING JUNCTIONS
COMMUNICATING JUNCTIONS
ANCHORING JUNCTIONS
Contd
The specialized junctions may be further classified as follows:
OCCLUDING JUNCTIONS
Gap junctions
ANCHORING JUNCTIONS
Adhesive junctions Cell to cell Zonula adherens(adheren junctions) Macula adherens (desmosome)
Cell to matrix Focal adhesion Hemidesmosomes
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Contd
The various junctions can be defined as: ADHERENS JUNCTIONS : A type of intercellular junction that links cell membranes and cytoskeletal elements within and between cells, connecting adjacent cells mechanically. Transmembrane proteins called cadherins link the neighbouring cells and connect to actin filaments. TIGHT JUNCTIONS :An intercellular junction at which adjacent plasma membranes are joined tightly together by interlinked rows of integral membrane proteins, limiting or eliminating the intercellular passage of molecules.
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strand act independently from others. Each strand is formed from a row of transmembrane protein. Transmembrane protein called claudin joins plasma membrane of 2 cells.
Contd
GAP JUNCTIONS : narrowed portion of intercellular space containing channels. In electrically excitable tissues, these gap junctions transmit electrical impulses via ionic currents and are known as electrotonic synapses. DESMOSOMES: (macula adherences)
It is a cell structure specialized for cell to cell adhesion
Found in simple & stratified sq. epithelium.
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DESMOSOMES
2 cells ,consisting of dense plate in each adjacent cells seprated by a thin layer of extracellular material.
Desmosomes link 2 cells together.
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HEMIDESMOSOMES:. Small Rivet like structure on the inner basal surface of keratinocyte in the epidermis of
skin. Forms the site of attachment between the basal surface of the cell and the basement membrane.
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TIGHT JUNCTIONS
Tight junctions, or zonula occludens:
closely associated areas of two cells forming a a tight belt like adhesive seal that
selectively limits the diffusion of water, ions & larger solutes as well as migration of cells. It is a type of junctional complex. Thus, separateing the interior of body from the external world.
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In electron microscopic structure of thin sections of tight junctions the plasma membrane of adjacent cells appear to fuse together in a series of one or more contacts.
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A.Preliminary model of tight junction structure with claudin linking the two membranes together & peripheral protein zo-1 linking the cytoplasmic tail of claudin to actin filaments. B-C. transmembrane topology of claudin & occludin.
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Tight junctions
Extracellular domains of claudins form rows of pores along tight
junction.
Each claudin has a unique selectivity for cations or anions. At the zona occludens the membranes of adjoining cells converge
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Contd
The cytoplasmic tails of claudins
interact with numerous proteins with roles as scaffolds and in actin binding, signalling and cell polarity.
Tight junctions are the barrier that
segregates different pumps, carriers, receptors and lipids in the apical and basolateral domains of plasma membrane.
Cell attachment is stronger at zonula occludens.
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Tight junctions
They may be tight as in distal convoluted tubule of
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Tight junctions
Several bacterial toxins also effects tight junction barrier.
These toxins disrupts tight junction barrier ,thus breaking
selective defects.
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Adherens junctions and Desmosomes are two types of adhesive junctions using homophilic interactions of cadherins to bind epithelial cells to adjacent cells.
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ADHERENS JUNCTIONS
The zonula adherens is a band like specialization of the
membrane and cytoplasm that encircles the apex of adjoining cells and strongly bonds the cells together.
In this junction the opposing membranes are 15 20 nm
apart .
It is a major site of epithelial cell cohesion.
adherent junction
Cytoplasmic actin filaments bind adherens junctions.
Homophilic interactions between densely clustered E-cadherens
(the epithelial transmembranic adhesive protein) bind adjacent cells together at adherens junctions.
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catenin(cytoplasmic adapter protein) and Plakoglobin (desmosomal cytoplasmic adapter protein) bind the cytoplasmic domains of E-cadherin.
adherent junction
An another cytoplasmic adapter protein, -catenin, binds
adherent junction
allow epithelial cells to establish polarity with proteins and lipids in plasma membranes.
Zonula adherens is the major site for cell cohesion.
It stabilizes the surface of epithelia. The junctions and polarity determine the orientation of mitotic
spindle and the plane of division . This allows for asymmetrical division of stem cells (stratified epithelium).
adherens junction
In mature columnar epithelia a belt like adherens junction called
zonula adherens encircles the cell near the apical surface thus maintaining physical integrity of the epithelium.
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DESMOSOMES
Desmos means bound , Soma means body.
cells.
These junctions are small disk shaped spot welds
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desmosomes
Cellular
desmosomes
are
mediated
by
Desmogleins
Desmocollins.
Plakoglobin also called gamma- catenin. Molecular composition of desmosomes vary in particular tissues
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desmosomes
Desmosomes are site for attachment, structural ability of
desmosomes.
The devlopment of animal tissues depends on desmosomes &
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GAP JUNCTIONS
Gap junctions are plaque that contain large intercellular
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gap junctions
Connexin are named by their molecular weight.
Found exclusively in chordates.
Gap junction communication is conditional It depends on: Number of channels Fraction that are open or closed
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gap junctions
Plants lacks gap junctions
Cells in plant tissues maintain continuity through
plasmodesmata.
Molecules smaller than 1kd diffuse freely through
plasmodesmata.
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gap junctions
Oleamide- fatty acid amide produced by the brain , blocks gap
endothelial cells.
Cells in most metazoans communicate by gap junctions.
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gap junctions
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HEMIDESMOSOMES Hemidesmosome
s
Basal Cells
Tonofilaments
Lamina lucida Lamina densa Anchoring filaments Anchoring fibrils
Proteoglycan s
Type IV collagen
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HEMIDESMOSOMES
Hemidesmosomes are adhesive
junction that link cytoplasmic filaments to basal lamina. Adhesion to extracellular matrix is different from intercellular adhesion because integrins provide transmembrane link between cytoskeleton and extracellular matrix.
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Contd
Two transmembrane proteins mainly found in hemidemosomes
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Contd
STRUCTURE OF HEMIDESMOSOME:
Adhesive protein INTEGRIN Cytoplasmic proteins PECTIN, BP 180 Cytoskeletal element - INTERMEDIATE FILAMENTS Target molecule - LAMININ.
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CELLULAR ADHESION
PRINCIPLES OF CELLULAR ADHESION
1.
expressing plasma membrane receptors with limited ligand binding activity. For example: Endothelial cells produce E selectin only when stimulated by inflammatory hormones
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Contd
2. Second principle of adhesion Many adhesion proteins bind one main ligand and many ligands bind a single type of receptor, for example Most cadherins bind to themselves, such homophilic interactions require Ca+2 ions.
3. Third principle of adhesion It states that cell modulate adhesion by controlling the surface density, state of aggregation and state of activation of their adhesion receptors.
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Contd
4. Fourth principle of adhesion The rates of ligand binding and dissociation are important determinants of cellular adhesion. Many cell surface adhesion proteins bind their ligands weakly in comparison with other specific macromolecules. 5. Fifth principle of adhesion Many adhesion proteins interact with the cytoskeleton inside the cell.
6. Sixth principle of adhesion Association of ligands with adhesion receptors can activate intracellular signal transduction pathways, leading to changes in gene expression, cell division, cell activation.
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protein.
Homophilic interactions of cadherins link epithelial and muscle
cells to adjacent cells at specialized junctions called adherens junctions and desmosomes.
The cytoplasmic domains of cadherin junctions interact with actin
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Cadherins are named according to their location or cells to which they are attached. for eg:
Epithelial tissue -E-cadherin
Nervous tissue - N-cadherin Placenta- P-cadherin
osteoblasts- O-cadherin
Kidney-K cadherin muscle -M-cadherin
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SIGNIFICANCE OF CADHERIN
Cadherins and catenins participate in transduction of
cell
proliferation,
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cadherin
Loss of E- cadherin expression also promotes tumour metastasis
to lymph nodes and higher frequency of blood- borne (bone and lung) distant metastasis.
Loss of E -cadherin can contribute to transition from benign to
malignant tumors.
Genetic defects in E-cadherin causes stomach cancer. Epithelial structures are not formed if cadherin expression is
impaired.
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receptors as some bind to several protein ligands & many matrix molecule bind to 1 integrin.
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Figure
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integrins
on ligands.
With the exception of red blood cells integrins are
integrins interact directly or indirectly with a remarkable variety of signaling & structural protien.
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domain.
The lectin domain sits at the end of a rod shaped projection that is
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selectin
Bonds between selectins and their mucin ligands have high tensile
endothelium, brief lifetime of this bond allows rolling motion over the surface of endothelium.
Inflammatry mediators also regulate selectin in several ways. How?
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years. Many histochemical and special stains can be carried out on formalin-fixed, paraffin embedded material .
The periodic acid- Schiff (PAS) stain demonstrates the
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Contd
The PAS reaction consists of oxidation of adjacent hydroxyl
groups in 1,2 glycols to aldehydes and staining of aldehydes with fuchsin sulfuric acid.
The PAS reaction is of value is studying the thickening of
basement membrane
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IMMUNOFLUORESCENCE
Immunofluorescence is a method of determining the
location of antigen (or antibody) in a tissue section or smear by the pattern of fluorescence resulting when the specimen is exposed to the specific antibody (or antigen) labelled with a fluoro chrome.
Immunofluorescence
their antigen to target fluorescent dyes to specific biomolecule targets within a cell, and therefore allows visualisation of the distribution of the target molecule through the sample.
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Contd
Immunofluorescence
is a widely used example of immunostaining and is a specific example of immunohistochemistry that makes use of fluorophores to visualise the location of the antibodies
INDIRECT IMMUNOFLUORESCENCE
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DIRECT IMMUNOFLUORESCENCE
Direct
immunofluorescence testing has a valuable diagnostic role in autoimmune diseases and inflammatory mucocutaneous diseases including auto immune mediated blistering diseases.
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Contd
This technique has several advantages over the secondary
(or indirect) protocol below because of the direct conjugation of the antibody to the fluorophore.
This reduces the number of steps in the staining procedure,
is therefore faster, and can avoid some issues with antibody cross-reactivity or non-specificity, which can lead to increased background signal.
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INDIRECT IMMUNOFLURESCENCE
Secondary, or indirect, immunofluorescence uses two
antibodies; the first (the primary antibody) recognises the target molecule and binds to it, and the second (the secondary antibody), which carries the fluorophore, recognises the primary antibody and binds to it.
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Contd
Antibodies can be prepared by the known antigen into an animal
tissue section containing the antigen, the antibody binds specifically to that antigen.
The antigen-antibody complex is subsequently attached to a
second antibody, which is conjugated either by a fluorescent dye (rhodamine) or enzyme (peroxidase-antiperoxidase)
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Contd
The antigen antibody complexes are bound to a fluorescent dye
where the antigenic sites fluoresce against a dark background and are immediately photographed on high speed film.
The enzyme bound antigen- antibody complexes are further
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ELECTRON MICROGRAPHY
It is a technique where beam of electrons is
transmitted through an ultra thin specimen interacting with the specimen as it passes through.
An image is formed from the interaction of the
electrons transmitted through the specimen; the image is magnified and focused onto an imaging device, such as fluorscent screen, on a layer of photographic film, or to be detected by a sensor such as a CCD camera.
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ELECTRON MICROGRAPHY
immunohistochemistry is negative.
Using electron microscopy the identification of intercellular
bullosa.
For optimum results the fresh tissue must be fixed in
karnovsky medium (paraformaldehyde and glutaraldehyde) and stored in refrigerator until processing.
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IMMUNOHISTOCHEMISTRY
This technique is used to diagnose poorly differentiated
malignant tumours and lymphomas. They can also be beneficial in diagnosis of bullous diseases.
With refinement of technique
immunohistochemistry methods have achieved the same sensitivity for many antigens as direct immunofluorescence method.
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SUMMARY
Intercellular junctions are fundamental to the interactions between cells. Mucosal barrier integrity is maintained by the physical interactions of
In the heart, cell junctions form the low-resistance pathways for rapid
impulse conduction and propagation, enabling synchronous stimulation of myocyte contraction. creating a balance between osmotic gradients.
make up tissues are co-ordinated, enabling each tissue system to function as an integrated whole.
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Contd
s They are connected to each other via membrane-associated structures usually transmembrane proteins.
These membrane-associated structures help in cohesion and
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Contd
The defects in intercellular junctions results in various
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