Intercellular Junctions

Thomas D. Pollard MD , William C. Earnshaw PhD, FRS

, Jennifer Lippincott-Schwartz PhD and Graham T. Johnson MA,
PhD, CMI Cell Biology, Chapter 31, 543-553
• Describe the function and importance of the different types of cellular junctions
• Know what intracellular proteins function in each type of cellular junction
• Give examples of where each type of cellular junctions is located
• Be able to draw a cell with all the cellular junctions shown with the names of the
proteins on the membrane and cytoplasmic proteins.

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 Two types of junctions to bind to the extracellular matrix. Subsequent research
established the molecular architecture of these junctions, each based on a different
transmembrane protein:

• Adherens junctions: Transmembrane proteins called cadherins link neighboring cells

and connect to actin filaments in the cytoplasm.

• Desmosomes: Another type of cadherin links cells together and connects to

cytoplasmic intermediate filaments.

• Tight junctions: Transmembrane proteins called claudins join the plasma membranes
of two cells to create a barrier that limits diffusion of ions and solutes between the cells
and molecules between apical and basolateral domains of the plasma membrane.

• Gap junctions: Transmembrane proteins called connexins form channels for small
molecules to move between the cytoplasms of neighboring cells.

• Hemidesmosomes: Integrins connect cytoplasmic intermediate filaments to the basal

lamina across the plasma membrane.

• Focal adhesions: Integrins associated with cytoplasmic actin filaments adhere to the
extracellular matrix.

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TABLE 31.1
Molecular Components of Cell-Cell and Cell-Matrix Junctions

Junction Target Adhesive Cytoplasmic Cytoskeletal

Molecule Protein Proteins Filaments
Sealing of the Extracellular Space
Tight junction Claudin Claudin ZO-1, ZO-2, Actin
cingulin,
spectrin
Communication Between Cells
Gap junction Connexin Connexin ZO-1, drebrin Actin
Adhesion to Other Cells
Zonula adherens Cadherin Cadherin Catenins, Actin
plakoglobin
Desmosome Desmoglein Desmoglein Plakoglobin, Intermediate
Desmocollin Desmocollin desmoplakin
Adhesion to the Extracellular Matrix
Hemidesmosome Laminin Integrin Plectin, BP 180 Intermediate
Focal contact Fibronectin Integrin Talin, vinculin, Actin
α-actinin

FIGURE 31.1

LIGHT AND ELECTRON MICROGRAPHS OF JUNCTIONS.

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A, Desmosomes. Left, Light micrograph of a section of skin showing numerous desmosomes as pink dots between the
cells. Right, Electron micrograph of a thin section of skin showing desmosomes. B, Light micrograph of a section of
intestinal epithelium stained with hematoxylin and eosin, showing the junctional complex (also called “terminal bars”)
as bright pink dots between the cells near their apex, just below the microvilli of the brush border. C, Electron
micrograph of a thin section of intestinal epithelial cells, showing the junctional complex consisting of a belt-like tight
junction (also called the zonula occludens), a belt-like adherens junction (also called the zonula adherens), and
desmosomes (also called the macula adherens), all in their characteristic relation to each other. The circumferential
tight junction seals the extracellular space. The zonula adherens is anchored to the actin cytoskeleton. Desmosomes
are attached to cytoplasmic intermediate filaments. D, Drawing showing the position of the junctional complex in the
cell and the locations of gap junctions, basal lamina, and hemidesmosomes.( A, Courtesy Don W. Fawcett, Harvard
Medical School, Boston, MA. C, Courtesy Marilyn Farquhar, University of California, San Diego.)

Each tissue uses a selection of junctions suited to its physiological functions. Columnar
epithelial cells in the intestine interact with their neighbors using all four types of
intercellular junctions ( Fig. 31.1B–D ). Belt-like tight junctions and adherens junctions
encircle the apex of the cell. Desmosomes and gap junctions form patch-like lateral
connections between the cells. Hemidesmosomes anchor the cells to the basal lamina.
Stratified epithelial cells in the skin ( Fig. 31.1A ) use desmosomes and intermediate
filaments ( Fig. 31.1B ) to resist mechanical forces but also interact via claudins and
adherens junctions. Desmosomes and adherens junctions link muscle cells to the
surrounding basal lamina (see Fig. 29.17C ). Gap junctions connect heart and smooth
muscle cells, but not skeletal muscle cells. Most nerve cells communicate chemically, but
some use gap junctions for electrical communication.

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Tight Junctions
Tight junctions form a belt-like adhesive seal that selectively limits the diffusion of water,
ions, and larger solutes between epithelial cells ( Fig. 31.2 ). This allows epithelia to
separate the interior of the body from the external world. Tight junctions also define the
boundary between the biochemically distinct apical and basolateral domains of the
plasma membrane of polarized epithelial cells.

FIGURE 31.2 EPITHELIAL TIGHT JUNCTIONS

A, Electron micrograph of a thin section of endothelial
cells, showing a point of contact between the plasma
membranes at a tight junction (arrow). B, Electron
micrograph of a replica of a freeze-fractured cell. This
method exposes proteins within the lipid bilayer and
reveals strands aligned along the points of contact
between the plasma membranes. C, Drawing showing
the strands of transmembrane proteins at points of
contact. ( A, Courtesy George Palade, University of
California, San Diego. B, Courtesy Don W. Fawcett,
Harvard Medical School, Boston, MA.)
.

Tight junctions were first recognized in electron micrographs of thin sections as places
where the plasma membranes of adjacent cells appear to fuse together in one or more
contacts ( Fig. 31.2 ).

Transmembrane proteins forming the strands observed by freeze-fracture were difficult

to identify until investigators found an antibody that bound to the cytoplasmic side of the
plasma membrane at tight junctions. They used this antibody to isolate an integral
membrane protein and named it occludin. However, mice lacking their single occludin
gene survive with normal tight junctions. Subsequently, these scientists
discovered claudins, the main structural proteins of tight junction strands ( Fig. 31.3A ).

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FIGURE 31.3

STRUCTURE OF TIGHT JUNCTIONS. A, Ribbon diagram of the transmembrane domains of claudin-

15. B, Interactions between the molecules in crystals of claudin-15 that are thought to represent contacts in tight
junctions. C, Model of tight junction structure with claudins linking the two membranes together and peripheral protein
ZO-1 linking the cytoplasmic tail of claudins to actin filaments. (For reference, see Protein Data Bank
[PDB; www.rcsb.org ] file 4P70 and Suzuki H, Nishizawa T, Tani K, et al. Crystal structure of a claudin provides
insight into the architecture of tight junctions. Science . 2014;344:304–307.)

The barrier in the membrane bilayer: Intimate lateral interactions of claudins within the
lipid bilayer ( Fig. 31.3B ) block diffusion of lipids and proteins in the plane of the
membrane. This barrier separates different pumps, carriers, receptors and lipids in the
apical and basolateral domains of the plasma membrane.

The barrier between cells: The small extracellular domains of claudins interact with their
neighbors in intramembrane strands and with claudins in similar strands on adjacent cells
to make barriers interrupted by rows of pores. These pores block diffusion of solutes
larger than approximately 1 nm in diameter, but selectively allow the passage of small
cations or anions. Most tight junctions are more permeable to cations than to anions.
Permeability in the two directions across the junction is identical.

Adapter proteins link tight junctions to the cytoskeleton. These adapters are called ZO-1,
ZO-2, and ZO-3 . They interact with the long C-terminal cytoplasmic tail of claudin and
JAM (junctional adhesion molecule) in the membrane. In the cytoplasm they interact with
actin filaments, a small guanosine triphosphatase (GTPase) and other proteins that
regulate actin polymerization.

These two barrier functions of tight junctions set up the two conditions required for many
physiological processes. The actions of pumps, carriers, and channels located
selectively in the apical or basolateral domains of the plasma membrane allow polarized
cells to create different extracellular environments on the two sides of the epithelium.

Maintaining these environments depends on the selective permeability across the

epithelium through the extracellular pores of tight junctions. For example, tight junctions
are essential for intestinal epithelial cells to take up nutrients from the lumen of the
intestine and transport them into the extracellular space beneath the

Several bacterial toxins affect the tight junction barrier. The ZO-toxin of Vibrio
cholerae induces diarrhea by loosening tight junctions, independent of the classic
cholera toxin, which induces secretion of salt and water. Helicobacter pylori injects a
protein toxin into the cells lining the stomach. This toxin disrupts tight junctions,
breaking the barrier that protects the underlying tissues and predisposing to ulcers.

Mutations of human claudin genes cause highly selective defects in epithelial barriers.
One example is reduced ability of the kidney to reabsorb potassium (claudin-16).
Another is deafness due to loss of ion gradients in the inner ear (claudin-14).

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Gap Junctions
The idea that channels might couple cells arose relatively late, because
electrophysiological experiments on nerves and skeletal muscles reinforced the
widespread belief that cells were autonomous. However, nerve and skeletal muscle cells
later turned out to be exceptions to the general principle that cells in animal tissues
communicate with each other by gap junctions.

junctions transmit action potentials from one cell to the next without the delay required
for secretion and reception of a chemical transmitter were found to connect heart
muscle cells

Gap junctions are plaques composed of large intercellular channels that connect the
cytoplasms of a pair of cells. These plaques exclude other transmembrane proteins and
contain a few to thousands of channels. Half channels in each membrane
called connexons are each formed from six protein subunits, named connexins ( Fig.
31.7 ).

FIGURE 31.6
LIGHT AND ELECTRON MICROGRAPHS OF GAP JUNCTIONS.
A, Thin section of embedded cells, showing the closely apposed membranes of adjacent cells
separated by a gap of 2 nm. B, Replica of a freeze-fractured cell, showing an irregular array of
particles exposed in the plane of the lipid bilayer. C, Fluorescence micrograph of a gap junction
plaque (red and green) between cultured HeLa (Henrietta Lacks) cells expressing connexin-43 with
a tetracysteine peptide tag. The cells were first exposed to a green fluorescent dye that binds tightly
to the tetracysteine tag and then, after 4 hours of growth without the green dye, the same cells were
incubated with a second red fluorescent dye that binds to the tetracysteine tag on newly synthesized
connexin-43. The older central part of this plaque is green. The newer peripheral regions of the
plaque are red. D, Negative staining of an isolated gap junction reveals the intercellular connexon
channels packed together in a regular, two-dimensional array. Each connexon has a central channel
filled with stain.
( A–B and D, Courtesy Don W. Fawcett, Harvard Medical School, Boston, MA; from the work of N.B.
Gilula, Scripps Research Institute, La Jolla, CA. C, Courtesy Mark Ellisman, University of California,
San Diego and from Gaietta G, Deernick TJ, Adams SR, et al: Multicolor and electron microscopic
imaging of connexin trafficking. Science . 2002;296:503–507.)

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Gap Junctions in Disease
Point mutations in connexin genes cause remarkably specific defects in humans (Table
31.2 ), considering that most connexins are expressed in several tissues. Recessive
mutations in the connexin-26 gene are the most common causes of inherited
human deafness. As many as 1 in 30 people are carriers, and their mutations may
contribute to hearing loss late in life. Connexin-26 participates in the transport of K + in
the epithelia supporting the sensory hair cells in the ear.

Adherens Junctions
Adherens junctions use homophilic (like-to-like) interactions of E-cadherins to bind
epithelial cells to their neighbors. Adherens junctions are essential for viability from the
earliest stages of animal embryonic development. In mature epithelia, a belt-like adherens
junction, called the zonula adherens, encircles the cells near their apical surface and
maintains the physical integrity of the epithelium. Adherens junctions also anchor muscle
cells to the extracellular matrix.

Adherens junctions can transmit mechanical forces between cells and reinforce tissues,
because the cytoplasmic domains of the E-cadherins are linked to the actin cytoskeleton.
Adapter proteins connect cadherins to actin filaments and signaling proteins including
guanine nucleotide exchange proteins for the Rho-family GTPases that promote actin
assembly and force generation by myosin. The adapter proteins include β-catenin, a
related protein called plakoglobin, p120-catenin, and the actin-binding protein α-catenin.
Moderate physical forces stabilize this link from the cadherin tail through β-catenin and
α-catenin to actin filaments.

Adherens junctions are the first connections established within developing sheets of
epithelial cells. Contact begins when cadherins on the tips of filopodia engage partner
cadherins of the same type on another cell. The contact spreads laterally as more
cadherins are recruited along with associated actin filaments, as illustrated by dorsal
closure of the ectoderm by Drosophila embryos . These pioneering adherens junctions
eventually allow like cells to associate in epithelial sheets (see Fig. 30.8 ) and to influence
the maturation of the epithelium. Adherens junctions are a prerequisite for the assembly
of tight junctions that allow epithelial cells to establish polarity with different proteins and
lipids in the apical and basal plasma membranes. The shapes of cells in epithelial sheets
depend on Rho family GTPases and protein kinases associated with the adherens
junction, which regulate the assembly and contraction of the associated actin
cytoskeleton.

The junctions and polarity of the cells determine the orientation of the mitotic spindle
and the plane of division. This allows for asymmetrical division of stem cells, such as
those at the base of stratified epithelia

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Desmosomes

Desmosomes ( desmos = “bound,” soma = “body”) use cadherins to provide strong

adhesions reinforced by intermediate filaments between epithelial and muscle cells. In
epithelia, these junctions are small, disk-shaped, “spot welds” between adjacent cells.
Desmosomes in the heart are more complicated because they are mixed with adherens
junctions (see Fig. 39.18 ).

Two families of desmosomal cadherins, named desmogleins and desmocollins ,

mediate cellular adhesion at desmosomes.

Desmosomal cadherins connect to cytoplasmic intermediate filaments via adapter

proteins analogous to those that connect adherens junction cadherins to actin filaments.
Two proteins related to β-catenin, plakoglobin and plakophilin , bind to cytoplasmic
domain of desmosomal cadherins and form a physical link to desmoplakin, a dimeric
protein related to plectin. The C-terminus of desmoplakin binds directly to the N-terminal,
nonhelical domains of epidermal keratin intermediate filaments. Mutations in this part
of epidermal keratins cause blistering skin diseases by compromising the integrity of
desmosomes (see Fig. 35.6 ).

The development of animal tissues depends on desmosomes. Loss-of-function mutations

can lead to mechanical failures. For example, mutations in the plakoglobin gene can be
lethal in mice and humans during embryogenesis, owing to disruption of the heart.
Similarly, mutations in the desmoplakin gene cause skin and cardiac defects that can be
fatal. Desmosomal proteins also participate in signal transduction. Loss of desmosomes
is associated with the spread of epithelial cancer cells.

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Adhesion to the Extracellular Matrix:
Hemidesmosomes and Focal Contacts
Adhesion to the extracellular matrix is fundamentally different from intercellular adhesion
because integrins, rather than homophilic interactions of cadherins, provide the
transmembrane link between the cytoskeleton and ligands in the extracellular matrix
(see Fig. 30.9 ). At focal contacts and related assemblies, transmembrane integrins link
cytoplasmic actin filaments to the extracellular matrix (see Fig. 30.11 ).

Hemidesmosomes are another type of integrin-based adhesive junction that links

cytoplasmic intermediate filaments to the basal lamina. The morphologic resemblance of
hemidesmosomes to half of a conventional desmosome belies the fact that they are
fundamentally different at the molecular level ( Fig. 31.8C ). Like desmosomes,
hemidesmosomes have a dense plaque on the cytoplasmic surface of the plasma
membrane that anchors loops of intermediate filaments. The similarity ends there.

FIGURE 31.8
COMPARISON OF ADHERENS JUNCTION, DESMOSOME, AND HEMIDESMOSOME.
Top, Electron micrographs of thin sections. Bottom, Molecular models. A, Adherens junction.
Electron micrograph from the intestinal epithelium. E-cadherins link two cells together. β-Catenin and
α-catenin link the cytoplasmic domain of E-cadherin to actin filaments. B, Desmosome. Two types of

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cadherins—desmoglein and desmocollin—link adjacent cells together. The central dense stratum
seen in the micrograph presumably corresponds to the interaction sites of the cadherins, although
accessory proteins may participate. Desmoplakin and other accessory proteins link the cadherins
and associated plakoglobin (related to catenin) to keratin intermediate filaments. Desmoplakin
molecules are shown extended to their full length in the middle drawing, whereas in desmosomes,
they must be kinked or folded (as shown in the upper drawing ) because the thickness of the
desmoplakin layer is half that expected from extended molecules. C, Hemidesmosome. Integrin
α 6 β 4 and type XVII collagen (also called BPAG2 [bullous pemphigoid antigen-2]) attach to the basal
lamina. Plectin, BP230, and BPAG1 (bullous pemphigoid antigen-1) link the membrane proteins to
keratin intermediate filaments. ( A–B, Micrographs courtesy Hilda Pasolli and Elaine Fuchs,
Rockefeller University, New York and from Perez-Moreno M, Jamora C, Fuchs E. Sticky business:
orchestrating cellular signals at adherens junctions. Cell . 2003;112:535–548. C, Micrograph
courtesy Jonathan Jones, Northwestern University, Chicago, IL.)

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