05Lecture, Sept 15/17 (Dr.

Tanentzapf)
Cell Adhesion
Gartner and Hiatt
3rd edition: Ch 5, pg. 85, pg. 90-102
4th edition: Ch 5, pg 99; pg. 103-118
Optional: Alberts' Molecular Biology of the Cell - Ch 19 (for a number of the figures in the
lecture powerpoint)

Four Basic Tissues

1) Epithelium
2) Connective Tissue
3) Muscle
4) Nervous tissue
• Tissues are formed when cells become arranged in complex 3-dimensaional structures. “Cell
junctions” facilitate the formation of tissues by linking individual cells with each other and the
surrounding extracellular matrix (ECM)

There are three 'functional' types of cell junctions:

1. Anchoring - mechanically attach cells to other cells or the ECM
2. Occluding - seal the contacts between neighboring cells to create isolated compartments
3. Channel-forming/communicating - form channels between cells for communication/allow
chemical and electrical signals to pass from cell to cell

All types of cell junctions are multi-protein complexes containing 3 main types of proteins
(ie. they have a common structural theme):
1. Transmembrane adhesion receptor proteins: These are integral membrane receptors that span
the cell membrane to connect the inside and outside environment of the cell. The outer
extracellular region attaches to other adhesion proteins on neighbouring cells (i.e. function in
cell-cell adhesion) or extracellular matrix molecules (i.e. function in cell-ECM adhesion). Their
inner intracellular regions attach to cytoplasmic 'adapter' proteins and connect to other proteins
like cytoskeletal linkers and cytoskeletal proteins themselves (eg. actin). Transmembrane
proteins are classified by how many times they cross the transmembrane region: single pass, two
pass, three pass transmembrane protein, etc.
2. Adapter proteins: They bind to the adhesion complex at the membrane, recruit additional
components to the adhesion complex and regulate the adhesion complex.
3. Cytoskeletal linkers: They physically link the adhesion complex proteins to the cytoskeleton.

Cell Junctions/adhesions in epithelia:

• Cell junctions are an especially prominent feature of epithelial cells. This lecture uses epithelial
cells as a model for exploring the function of cell junctions (for a description of epithelia
generally see pg. 84 of 3rd edition or pg. 99 of 4th edition; more on the specific types of epithelia
described in this chapter of the textbook in Lecture 07).
• Epithelial cells are polarized, meaning cell junctions are arranged in a particular way in the cell.
• Overall there are 6 types of junctions found in epithelial cells that we will discuss:
o Four Cell-Cell adhesions/junctions:
1. Tight Junctions (a type of Occluding/Sealing Junctions)
 2. Zonula Adherens (a type of Anchoring Junctions)
3. Desmosomes (a type of Anchoring Junctions)
4. Gap Junctions (a type of Channel Forming Junctions)
o Two Cell-ExtraCellular Matrix (ECM) adhesion/junctions (discussed in lecture 6):
1. Hemidesmosomes (Anchoring Junctions)
2. Focal Contacts (Anchoring Junctions)

Epithelial Cell Polarity and Cell Junctions:

• What is epithelial polarity? Positional asymmetry (eg. regional differences) within the
cell.
• In epithelia this is demarcated by three domains (eg. regions of the cell) based on their
position along the various surfaces of the cells. The cell junctions are characteristically
located in the various polarized 'domains' of the cells:
1. Apical domain - this is the free, unattached plasma membrane region that faces an open
space (eg. ‘outward’) that may be air-filled (eg. parts of the respiratory tract) or fluid-
filled (eg. blood).
2. Lateral domain - this is the membrane region that is in close contact with neighbouring
cells within the epithelium
3. Basal domain - this is the domain that is attached to extracellular matrix that often faces
underlying connective tissues (eg. 'inward')
• junctions are arranged in specific locations along the lateral domain (from apical side to
the basal side: tight junctions à adherens junctions à desmosomes à gap junctions)

Functional requirements for anchoring junctions

• Three common features of cell junctions increase their stability and strength given how small the
junction is to the very large size of the cell (in comparative terms):
1. Made up of multi-protein complexes
2. 'Clustering' of the transmembrane adhesion proteins in the plasma membrane - the
combined strength of multiple bond interactions increases overall strength of the junction
that binds to either other cells or the extracellular matrix
3. The clustered adhesion proteins link to the cytoskeletal network inside the cell which
produces a large scale/distributed tension-bearing protein interaction network that runs
throughout the tissue.

How can we visualize junctions?

One great way to visualize junctions is using a histological technique called freeze fracture
microscopy:

• Freeze fracture electron microscopy: in which carbon/platinum-coated 'casts' are made of

frozen membrane surfaces, make it possible to view adhesion protein clusters embedded in
the membrane as well as their associated adapter proteins and linker proteins (i.e. so that
the multiprotein junctional complexes can be imaged).

Please note: For examination purposes in this course you will not have to distinguish between
'E' and 'P' faces on a freeze fracture electron micrograph
 We will discuss 4 Specific Types Cell-Cell Junctions
• Cell-cell junctions form between cells
• Specific cell-cell junctions are classified based on their molecular structure and their function
• Described below in order starting from apical to basal in an epithelium formed by a single layer
of polarized epithelial cells. (a 'simple' epithelium; will be defined in Lecture 9)

1. Tight Junctions/Zonula Occludens ('occluding/sealing' junction)

• 'Zonula' = belt that goes all the way around the entire circumference of the cell near the apical
membrane domain
• Tight junctions are located very near the apical domain of polarized epithelial cells and they join
neighbouring cells very closely together.
• Formed by strands of interacting transmembrane proteins observable by freeze-fracture electron
microscopy. Two core tight junction adhesion receptor proteins are Claudin and Occludin.
Claudins are required for tight junction formation and occludins are required for barrier function
of the junction. Binding to the same type of molecule on neighbouring cells (eg. Claudin-Claudin
or Occludin-Occludin interactions between two neighbouring cells that are held 'tightly' together).
• Appears like cobblestone path in EM image OR
• Analogous to sewing machine stitching two pieces of fabric together
• Barrier permeability can be tested by using a dye/tracer that is added to either the apical or basal
side of the epithelium and observing whether the dye/tracer diffuses past the tight junction
• The barrier function of the tight junctions controls diffusion of material between cells (eg. along
the lateral membranes). This is very important physiologically as it sets up epithelia as 'barriers'
where transport of molecules (eg. glucose for example, across the intestinal epithelium) can be
highly regulated by the cells based on polarized insertion of specific plasma membrane
transporters in the apical and basal domains of the intestinal epithelial cells (the example used in
the lecture was glucose transport, the same is true of many electrolytes including sodium and
chloride ions (we will return to this concept in the Gastrointestinal section of the course).
• Clinical/disease relevance: Loss of the tight junction adhesion protein Claudin-16 (due to
mutation of one of many claudin genes) disrupts the ability of kidney epithelial tubules to
efficiently transport electrolytes out of the urinary filtrate. As a result, calcified deposits form
within the kidneys (page 12 in lecture powerpoint: white dots in the CT), which can eventually
lead to blockage and kidney failure (we will return to this concept in the Urinary section of the
course)

2. Adherens Junctions/ Zonula Adherens (anchoring junction)

• Form strong continuous adhesion belts around the circumference of the entire cell on the lateral
domain just basal to tight junctions.
• Important for the formation of 2-dimensional sheets of epithelial cells that line our body
compartments.
• The adhesion receptor proteins are composed of single-pass transmembrane adhesion proteins
called classical cadherins
• Clustering of cadherins:
1. At low calcium levels two cadherin molecules interact with each other on the surface of the
same cell to form cis-homodimers
 • Cis-homodimerization occurs in the ER before it hits the plasma
membrane
2. Calcium-binding to the extracellular domains of the cis-homodimers causes them to
undergo a conformational change and 'straighten'
2. Cadherin straightening promotes "trans" homodimerization, which means that a cadherin
cis-homodimer pair on one cell then binds another cadherin cis-homodimer pair located on
the surface of a neighbouring cell to initiate adhesion between cells (i.e. now have a mini-
adhesive cluster of four cadherins)
3. These trans-bound cadherin mini-clusters then associate with each other to form large
adhesive clusters in the plane of the membrane (eg. this strengthens and stabilizes
adhesion).
• Cadherin clusters then link to the actin cytoskeleton via intracellular anchoring complex
containing adapter/linker proteins called "catenins" (these interactions strengthen the adhesion
between cells).
• Clinical relevance: Cadherins are 'tumor suppressors' in epithelial tissues. Specifically, a loss of
cadherins removes the 'suppression' than can contribute to the formation of, and the metastatic
progression of, cancers that arise in epithelium ('carcinomas'). A hallmark of a carcinoma is the
loss of epithelial structure, which is often associated with the absence of epithelial cadherins (by
mutation and/or transcriptional repression). This facilitates a loss of cell cycle control that
causes the initial 'primary' tumor to form. The loss of cadherins can also facilitate the
dissociation of tumor cells from each other which helps initiate 'tumor invasion' which is the first
step in metastasis that ultimately leads to the progressive formation of tumors at secondary sites
that is a major cause tumor mortality. The process of carcinoma-associated cell dissociation and
invasion has many things in common with the epithelial-to-mesenchymal transition [EMT] that
occurs when mesenchymal cells to break away from the epiblast and invade the space between
the developing ectoderm and endoderm during gastrulation).

3. Desmosomes/Macula Adherens (anchoring junction)

• Macula = spot; these are small spotlike-junctions that further strengthen the adhesion between
cells in tissues that are subject to high tensional/mechanical forces (i.e. they are very prominent
in the epithelium of the skin - we will return to this concept in the Skin lecture). These junctions
form along the entire lateral domain of cell types in tissues that are exposed to tensile
forces/mechanical stress.
• Desmosomes are junctions that are based on adhesions between cadherin-like receptors called
Desmocollin (Dsc) and Desmoglein (Dsg), which bind to cytoskeletal linkers to connect with the
keratin family of intermediate filaments (eg. 'KIF' on the TEM image on page 17 of the
powerpoint/PDF). Keratin intermediate filaments are stronger than the actin filaments found in
adherens junctions.
• Clinical/disease relevance: Pemphigus vulgaris - A mutational loss or functional disruption of
desmoglein causes a loss of adhesion between layers within the epithelium of skin, resulting in
uncontrolled skin blistering.

4. Gap Junctions (communicating junction)

• Major function is to allow communication and sharing of small molecules (eg. ions, signal
transduction membranes and some nutrients) between neighboring cells (in epithelial tissues and
other tissues, most prominently in cardiac myocytes (which will be discussed in Muscle lectures).
 • In TEM, you can see a plaque of gap junctions between two cells (cells “kissing”)
• Connexins are the gap junction adhesion receptor proteins. Six connexin proteins cluster together
as a hexamer called a connexon in the plane of lateral membrane domain of one cell. One
connexon will then bind a similar hexamer/connexon in the lateral membrane domain of a
neighbouring cell. This forms a functional channel between the two neighbouring cells that can
be regulated (eg. can be opened or closed) and can pass small molecules between linked cells in
the tissue (biochemical communication).
• Clinical/disease relevance: Vohwinkel Syndrome (keratodema) - Loss of connexin-26 (one of
many isoforms of connexin) causes a disruption of communication between epidermal cells in the
skin. This causes the skin epithelial cells to hyperproliferate ('hyperplasia') and to overproduce
keratin. Thus the skin becomes abnormally thick and hard (keratodema).