CHAPTER 31
Intercellular Junctions
T he mechanical integrity of animal tissues such as
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 these interactions. Physical connec
tions from the extracellular matrix or adjacent cells
through these junctions and the associated cytoskeletal
filaments inside cells impart mechanical strength to
tissues.
Investigation of junctions began when microscopists
and physiologists recognized that epithelial and muscle
cells adhere to each other and the underlying extracel
lular matrix. They also discovered that some epithelia
form a tight barrier between the luminal surface and the
underlying tissue spaces. The physical basis of these
interactions became clear during the 1960s, when elec
tron micrographs of thin sections of vertebrate tissues
revealed four types of intercellular junctions that connect
the plasma membranes of adjacent cells (Table 31.1 and
Fig. 31.1) and 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 (see Fig. 30.5) 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 con
nexins form channels for small molecules to move
between the cytoplasms of neighboring cells.
Hemidesmosomes: Integrins (see Fig. 30.9) 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.
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. 31.1BD). 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. Hemidesmo
somes 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 mechani
cal 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 junc
tions for electrical communication.
Invertebrate animals assemble junctions from homolo
gous proteins but with different organization than the
junctional complex of vertebrate epithelia. Insect epithe
lia have apical adherens junctions and more basal septate
junctions built from claudins and cytoplasmic proteins
with sequence homology to the tight junction proteins
ZO-1 and ZO-2. Nematode epithelia have one type of
junction with adherens functions and claudins.
Tight Junctions
Tight junctions form a belt-like adhesive seal that selec
tively 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
543
544 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

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

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

A. Skin B. Epithelium C. Epithelium

Desmosome

Zonula occludens
D. Epithelium (tight junction)

Zonula occludens
(tight junction) Zonula adherens
Zonula adherens
Macula adherens
(desmosome)
Gap junction

Macula adherens
(desmosome)

Hemidesmosome
Basal lamina Gap junction

FIGURE 31.1 LIGHT AND ELECTRON MICROGRAPHS OF JUNCTIONS. 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.)

external world. Tight junctions also define the boundary
between the biochemically distinct apical and basolat-
eral domains of the plasma membrane of polarized
epithelial cells.
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). Freeze-fracture images
revealed that these contacts correspond to continuous
strands of intramembranous particles that form a branch
ing network in the plane of the lipid bilayer.
A B C Plasma
membrane
Strands of
claudins
Intercellular
space
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.)
N
A C
B
90
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, etal. Crystal structure of a claudin provides insight into
the architecture of tight junctions. Science. 2014;344:304307.)
CHAPTER 31 n Intercellular Junctions 545
Transmembrane proteins forming the strands observed
by freeze-fracture were difficult to identify until investi
gators found a monoclonal antibody that bound to the
cytoplasmic side of the plasma membrane at tight junc
tions. They used this antibody to isolate an integral
membrane protein and named it occludin. The amino
acid sequence of occludin suggested four transmem
brane strands and two hydrophobic extracellular loops.
However, mice lacking their single occludin gene survive
with normal tight junctions. Subsequently, these scien
tists discovered claudins, the main structural proteins
of tight junction strands (Fig. 31.3A). Humans have a
family of 27 homologous claudin genes that are expressed
selectively in various tissues.
Claudins consist of four highly conserved transmem
brane helices with two extracellular loops that fold into
a small -sheet and single helix. Close associations
between claudins form barriers to diffusion in the plane
of the membrane and between the cells.
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 differ
ent 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 1nm 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 direc
tions across the junction is identical.
Pores
ZO-2
Claudin
ZO-1
Actin
546 SECTION VIII n Cellular Adhesion and the Extracellular Matrix
Adapter proteins with PDZ protein-interaction domains
(see Fig. 25.10) 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 fila
ments, a small guanosine triphosphatase (GTPase) and
other proteins that regulate actin polymerization and the
adapter protein cingulin. ZO-2 and cingulin are specific
for tight junctions, but ZO-1 also associates with cadherins
in adherens junctions and connexins in gap junctions.
These two barrier functions of tight junctions set up
the two conditions required for many physiological pro
cesses. The actions of pumps, carriers, and channels
located selectively in the apical or basolateral domains of
the plasma membrane allow polarized cells to create dif
ferent 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
cells (see Fig. 17.2) and for other physiological processes
(see Figs. 17.3 and 17-4). Restricting the diffusion of
macromolecules across epithelia can regulate some types
of signaling. For instance, airway epithelial cells release
the growth factor heregulin from the apical surface, but
restrict its receptor tyrosine kinase erbB2 (see Fig. 24.5B
for a related receptor) to the basolateral surface. Thus,
the receptor is activated only if the epithelium is damaged
or the tight junctions compromised.
Sheets of epithelial cells vary by several orders of
magnitude in the quality of the seal created by circum
ferential tight junctions between adjacent cells. The
tightness of the barrier to diffusion of ions in the extra
cellular space determines the electrical resistance across
the epithelium and depends on the claudin isoform and
the number and continuity of the strands. Epithelia also
appear to differ in the ability of water to flow through
tight junctions. Extremely tight barriers with many
strands and distinct claudin forms are found where epi
thelia must maintain high ion gradients, such as in the
distal tubules of the kidney, where urine is concentrated.
Leaky tight junctions with fewer strands and different
claudins are found where ion gradients across epithelia
are small but a barrier is required for large solutes, pro
teins, and leukocytes (eg, in most blood vessels).
Intracellular and extracellular factors regulate the
transepithelial barrier established by tight junctions.
These regulators include hormones such as vasopressin
and aldosterone and cytokines such as tumor necrosis
factor (see Fig. 24.9) that act through second messengers
(eg, Ca2+ and cyclic adenosine monophosphate [cAMP];
see Chapter 26), and effectors (eg, protein kinases A
and C; see Chapter 25). The mechanisms are not yet
well understood, but posttranslational modifications of
tight junctions might modulate their assembly. Tension
on associated actin filaments may physically open pas
sages through tight junctions. The metabolic state of
the cell also influences tight junctions; depletion of
adenosine triphosphate (ATP) causes tight junctions to
leak without destroying the barrier between the apical
and basolateral domains of the plasma membrane. White
blood cells migrating across epithelia from the blood
to the connective tissue, open tight junctions locally
without disrupting the tight seal across the epithelium
(see Fig. 30.13). A localized increase in cytoplasmic
Ca2+ in the epithelial cells is required to open the tight
junctions.
Several bacterial toxins affect the tight junction
barrier. The ZO-toxin of Vibrio cholerae induces diar
rhea 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 underly
ing tissues and predisposing to ulcers.
Mutations of human claudin genes cause highly selec
tive 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).
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 communi
cate with each other by gap junctions. Cells in plant
tissues also communicate with each other, but they use
direct cytoplasmic connections, called plasmodesmata,
rather than gap junctions (Box 31.1 and Fig. 31.4).
The first convincing evidence for direct electrical
communication between cells came around 1960
from electrophysiological experiments on the synapses
between giant axons and the motor neurons that drive
the flipper muscles of crayfish. These electrical syn-
apses transmit action potentials (see Fig. 17.6) directly
from one cell to the next without the delay required for
secretion and reception of a chemical transmitter (see
Fig. 17.10). Similar electrical junctions were subsequently
found to connect heart muscle cells (see Fig. 39.18).
Over the next decade, physiologists used microelec
trodes to establish that plasma membrane depolarization
of one cell can be transmitted with little resistance to
adjacent epithelial cells (Fig. 31.5B), although the ampli
tude of the response declined with distance. Similarly,
fluorescent molecules, radioactive tracers, and essential

BOX 31.1 Plasmodesmata
Most cells in plant tissues maintain cytoplasmic continuity
with their neighbors through plasmodesmata, channels
across the cell wall lined by plasma membrane across the
cell wall (Fig. 31.4). These connections form by incom
plete cytokinesis, but secondary plasmodesmata can form
independently. Plasmodesmata are essential for plant
viability.
A narrow tubule of modified endoplasmic reticulum
(ER) fills most of the pore and is linked to the surround
ing plasma membrane by a number of proteins. The
protein spokes connecting the ER and plasma membrane
are not yet well characterized, but candidates include
proteins that participate in membrane contact sites
in animals and fungi such as synaptotagmins (see
Chapter 21), synaptobrevin (see Chapter 21), lipid
exchange proteins (see Fig. 20.17), junctophilins, and
stromal interaction molecules (STIMs) (see Fig. 26.12).
Molecules smaller than about 1kD diffuse freely
through the narrow cylinder of cytoplasm in plasmodes
mata, but larger molecules, even nucleic acids pass
selectively through these channels. Constitutive diffusion
of small molecules allows exchange of metabolites and
hormones between cells. Regulated passage of larger
molecules, including double-stranded RNAs and proteins
such as transcription factors, allows developmental signals
to move between cells and tissues. Specialized viral
movement proteins allow some viruses to move their
whole genomes between cells.
Permeability varies among tissues and with physiologi
cal states and developmental stages. For example, all cells
in plant embryos are connected, whereas cells in some
adult tissues are isolated. Reversible deposition of callose
and other molecules in the cell wall surrounding plasmo
desmata regulates their diameter and permeability. Actin
filaments contribute to regulation of the pore size, but the
mechanism and signals controlling permeability are not
known.
Integral membrane proteins of the ER and Annulus
plasma membrane Desmotubule
ENDOPLASMIC
RETICULUM
CELL WALL
100 nm
FIGURE 31.4 A PLASMODESMATA CONNECTING TWO
PLANT CELLS. The plasma membrane is continuous between the
two cells. The space between the tubule of endoplasmic reticulum and
the surrounding plasma membrane allows molecules in the cytoplasm
to move between the two cells. ER, endoplasmic reticulum.
CHAPTER 31 n Intercellular Junctions 547
A Fluorescein
3x 4 5
2
1 3 4 5
2
1
Halothane
B
I1
V1
100 pA
I2
10 mV
V2
10 sec
C open
close
I1
10 pA
I2
2 sec
FIGURE 31.5 GAP JUNCTION PHYSIOLOGY. A, Drawing and
fluorescence micrograph, showing the movement of a tracer dye
between epithelial cells from the salivary gland of Chironomus. Cell 3
was injected with fluorescein (molecular weight: 330), which spread to
adjacent cells via gap junctions. BC, Electrical recordings from pairs
of cells coupled by gap junctions. B, Two cells (1 and 2) were voltage-
clamped (see the text that describes Fig. 17.6) and subjected alter-
nately to small depolarizing voltage changes (V1, V2). Being electrically
coupled, they responded with opposite currents (I1, I2). Transient
exposure to the anesthetic halothane (horizontal bar) closes most
of the channels, reducing the current in response to depolarization.
C, When the cells are held at a constant depolarizing voltage in the
presence of halothane, current records reveal the opening and closing
of individual gap junction channels as opposite step changes in
current. (A, From Lowenstein W. Junctional intercellular communica-
tion: the cell-to-cell membrane channel. Physiol Rev. 1981;61:829.
B and C, From Eghbali B, Kessler JA, Spray DC. Expression of gap
junction channels in communication-incompetent cells. Proc Natl Acad
Sci U S A. 1990;87:13281331.)
nutrients can pass from the cytoplasm of one cell to the
cytoplasm of neighboring cells.
Electron microscopy revealed that low-resistance
communication between cells is associated with the
presence of plasma membrane specializations that were
called gap junctions owing to the regular 2- to 4-nm
separation of the adjacent cell membranes (Fig. 31.6).
Gap junctions are plaques composed of large intercel-
lular channels that connect the cytoplasms of a pair of
cells. These plaques exclude other transmembrane pro
teins and contain a few to thousands of channels. Half
548 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

A B C D

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 2nm. 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. (AB 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, etal: Multicolor and electron microscopic imaging of connexin trafficking. Science.
2002;296:503507.)

channels in each membrane called connexons are each

formed from six protein subunits, named connexins
(Fig. 31.7).
Structure of Gap Junction Channels
A hexagonal ring of connexins forms a central aqueous
channel across the lipid bilayer and pairs with a con
nexon in an adjacent cell to connect their cytoplasms.
Four transmembrane -helices of each connexin subunit
span the lipid bilayer and two loops between the helices
form small extracellular domains (Fig. 31.7). Connexons
on adjacent cells dock to form a continuous pore
between the pair of cells. Tight interactions between the
subunits seal the pore and preclude leakage of ions out
of either cell.
The transmembrane pore begins with a funnel from
the cytoplasm that narrows to a diameter of 1.4nm,
larger than the pores of tetrameric S5-P-S6 ion channels
(see Fig. 16.5) or pentameric ligand-gated ion channels
(see Fig. 16.12). This pore passes hydrophilic molecules
up to approximately 1kD in size, including ions (to
establish electrochemical continuity between the cells),
second messengers (to establish a common network of
information), small peptides, and metabolites (to allow
sharing of resources). Connexon hemichannels (the ring
of six connexins in one plasma membrane) also open
infrequently for the nonspecific release of ions and
solutes as large as ATP from the cell.
Connexin Gene Families and Evolution
Humans have genes for 21 connexin isoforms, ranging
in size from 26 to 60kD. Connexins are named by
molecular weight; for instance, connexin-43 (Cx-43) is
the name for the 43-kD isoform. All connexins have the
conserved features required to form the connexon
channel but variable N- and C-terminal cytoplasmic
sequences. The various connexin isoforms make chan
nels that differ in their permeability and charge
selectivity.
Remarkably, gap junction genes seem to have arisen
more than once during evolution. Connexins are found
exclusively in chordates, while invertebrate gap junc
tions are composed of innexins (invertebrate connex
ins). Innexins have four transmembrane helices but
lack any sequence similarity to connexins and form
intercellular junctions from two octameric hemichan
nels. Vertebrates have a few genes related to innexins
called pannexins. Rather than forming gap junctions,
pannexins form plasma membrane channels that release
solutes including ATP from the cytoplasm. The ATP
activates seven-helix receptors (see Fig. 24.2) and chan
nels activated by adenine nucleotides (see Fig. 16.2) as
part of local signaling pathways in the immune and
nervous systems. Many animals have another channel,
calcium homeostasis modulator 1, that shares structural
features with connexins, pannexins, and innexins.
 CHAPTER 31 n Intercellular Junctions 549

A B D INTRA EXTRA INTRA

H1 CELLULAR CELLULAR CELLULAR
N
Conserved H2 38 56 aa
regions
H3
18 195 aa
H4 N-terminal
C
helix plug
EXTRACELLULAR CYTOPLASM

C E
Negatively
charged
Open patches

N-terminal
helix plug

Closed

FIGURE 31.7 STRUCTURE OF THE GAP JUNCTION CONNEXON CHANNEL. A, Drawing of gap junction connexons forming channels
between the cytoplasms of adjacent cells. B, Transmembrane topology of connexins. Four -helices cross the lipid bilayer. Conserved
residues (maroon) form the transmembrane and extracellular loops are required for channel assembly. Cytoplasmic loops between helix 2 and
helix 3 and the C-terminal tails vary in length among connexin isoforms. Removal of the C-terminal tail from connexin-43 alters its gating properties.
C, Diagram showing how the N-terminal -helices of Cx26 may form a plug that blocks the pore of the closed channel. DE, Crystal structure
of the connexin 26 the gap junction channel. CD, Top and side views of a ribbon diagram with each of the six subunits a different color. The
two extracellular loops of each subunit associate with the other half channel to span the 4-nm gap between the membranes. The upper panel
of D shows as a space-filling model cut through the middle of the transmembrane pore. (For reference, see PDB file 2ZW3 and Maeda S,
Nakagawa S, Suga M, etal. Structure of the connexin 26 gap junction channel at 3.5 A resolution. Nature. 2009;458:597602.)

However, the lack of sequence homology suggests that

this gene may have arisen separately.
Assembly of Gap Junctions
Connexin proteins turn over on a time scale of several
hours and are replaced by new connexons that assemble
in vesicles along the secretory pathway. New connexons
add around the periphery of gap junction plaques in the
plasma membrane and old connexons are removed from
the middle of plaques (Fig. 31.6C).
Many gap junctions are composed of one connexin
isoform and pass molecules equally well in both direc
tions. However, some connexons assemble from mix
tures of subunits creating hybrid gap junctions with
novel properties. Furthermore, some connexons can
pair with a different type of connexon on the neighbor
ing cell. Such hybrid channels may pass fluorescent
tracers more readily in one direction than the other or
react more sensitively to the transjunctional potential of
one polarity than the other. This might explain the asym
metrical coupling that is sometimes observed between
both excitable and nonexcitable cells, such as neuronal
gap junctions, which pass action potentials in one direc
tion but not the other.
Regulation of Gap Junction Permeability
Electrophysiological measurements showed that con
nexons alternate between open and closed states (Fig.
31.5C). The structural basis for this gating is not estab
lished definitively, but a plug-gating mechanism is plau
sible. In this model N-terminal helices can either form a
plug that blocks the pore or pull back against the wall
of the channel to create the 1.4-nm pore (Fig. 31.7E).
This would allow the two connexins to gate the pore
independently; both must be open to connect the two
cytoplasms. Cytoplasmic loops, C-terminal tails or the
extracellular loops may also contribute to gating.
The pores of gap junction channels are large enough
to pass all common inorganic ions as well as nucleotides,
amino acids, and even small peptides and RNAs. The
conductance of the open state depends on the connexin
isoform and varies from about 30 psec to 300 psec.
Given the permeability of gap junctions to relatively
large solutes, it is surprising that their conductance is in
the same range as narrower ligand- and voltage-gated ion
channels. Both the greater length and the arrangement
of charged residues lining the channel may contribute to
the low conductance of connexons.
550 SECTION VIII n Cellular Adhesion and the Extracellular Matrix
Gap junctional communication depends on both the
number of channels and the fraction of those channels
that is open or closed. The fraction of open channels is
usually less than 1.0; it is approximately 0.2 in heart and
as low as 0.01 in one nerve cell that was tested.
Many factors regulate the opening and closing of con
nexon channels. The transjunctional potential (ie, the
potential difference between the coupled cells) gates
most connexons, regardless of the plasma membrane
potentials of these cells. Like other voltage-gated chan
nels, individual transitions are fast, but the response to
potential changes, on the scale of seconds, is very slow
in comparison with other channels (see Fig. 16.7).
Unphysiological concentrations of cytoplasmic Ca2+ (100
to 500 M) and cytoplasmic acidification also close con
nexons. These effects of membrane potential, H+, and
Ca2+ allow cells to terminate communication with neigh
boring cells that are damaged (depolarizing the plasma
membrane and admitting high concentrations of Ca2+) or
metabolically compromised (allowing Ca2+ to leak out of
intracellular stores and acidifying the cytoplasm).
Chemicals also modulate gap junctions. Oleamide, a
fatty acid amide produced by the brain, blocks gap
junctional communication and induces sleep in animals.
Organic alcohols (heptanol and octanol) and general
anesthetics (halothane) can also close gap junction chan
nels reversibly (Fig. 31.5), but these agents are not spe
cific for gap junctions.
Signaling pathways control gap junction activity
through phosphorylation of numerous sites by several
kinases. For example, on a time scale of seconds, cAMP
activates protein kinase A, which phosphorylates the
C-terminal tail of some connexins, increasing or decreas
ing the fraction of open channels (depending on the
connexin isoform and the cell type). On a time scale
of hours cAMP also promotes the assembly of gap
junctions.
Physiological Functions of Gap Junctions
Cells in most metazoans communicate by gap junctions.
Coupled cells in vertebrates include epithelial cells of
the skin, endocrine glands, exocrine glands, gastrointes
tinal tract, and renal-urinary tract as well as smooth
muscle, cardiac muscle, bone, some neurons, and glial
cells. Epithelial cells can coordinate their activities with
their neighbors. This is used to synchronize the beats of
cilia (see Fig. 38.12C). Fragments of viral proteins can
spread from infected cells to neighboring cells, which
then become targets for cytotoxic T lymphocytes (see
Fig. 28.9). Gap junctions allow osteocytes buried deep
in bone to maintain a cellular supply line to acquire
nutrients from distant blood vessels (see Fig. 32.4).
Passage of action potentials between cardiac and smooth
muscle cells sets off waves of contraction (see Fig.
39.18). Electrical synapses between neurons can trans
mit action potentials at very high frequencies (>1000
TABLE 31.2 Phenotypes of Humans With Mutations
in Gap Junction Subunits
Connexin Phenotype
Cx-262 Dominant and recessive mutations with deafness;
skin disease
Cx-306 Recessive deafness; skin disease
Cx-313 Recessive deafness; skin disease
Cx-321 Point mutations, defective myelin, peripheral
nerve degeneration in X-linked Charcot-Marie-
Tooth disease; deafness
Cx-374 Female infertility, defect in communication of
granulosa cells with oocyte
Cx-405 Partial block of impulse conduction in heart
Cx-431 Deafness; many mutations may be lethal as in mice
Cx-463 Cataracts in lens of the eye
Cx-508 Cataracts in lens of the eye
Note: Mutations are homozygous loss of function mutations unless noted
otherwise. The nomenclature used here combines the Cx-molecular
mass in kD and molecular phylogeny -number systems.
per second). In some parts of the brain, gap junctions
also coordinate action potentials in groups of neurons.
Even white blood cells may form transient gap junctions
with endothelial cells.
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. Reces
sive 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 par
ticipates in the transport of K+ in the epithelia supporting
the sensory hair cells in the ear. Patients with one of
more than 100 different mutations in the connexin-32
gene can suffer from degeneration of the myelin
sheath around axons, an X-linked variant of Charcot-
Marie-Tooth disease. Many human tissues express
connexin-32, but the pathology is confined to myelin.
The stability of myelin might depend on intracellular gap
junctions between layers of the myelin sheath that
provide a pathway between the metabolically active cell
body and the deep layers of the sheath near the axon.
Defects in myelin membrane proteins cause other forms
of Charcot-Marie-Tooth disease.
Adherens Junctions
Adherens junctions use homophilic (like-to-like) interac
tions of E-cadherins (see Fig. 30.5) 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 (Fig. 31.1D) 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 cyto
plasmic 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 genera
tion by myosin (see Fig. 33.19F). The adapter proteins
include -catenin, a related protein called plakoglobin,
p120-catenin, and the actin-binding protein -catenin.
Moderate physical forces stabilize this link from the
cadherin tail through -catenin and -catenin to actin
filaments.
Adherens junctions are the first connections estab
lished 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 illus
trated by dorsal closure of the ectoderm by Drosophila
embryos (see Fig. 38.5). 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 mem
branes. 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 (see
Figs. 35.6 and 41.4).
Desmosomes
Desmosomes (desmos = bound, soma = body) use for binding DNA (see Fig. 30.7) and inhibiting the
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 des-
mogleins and desmocollins, mediate cellular adhesion
at desmosomes (see Fig. 30.5 and Table 30.2). The
most distal of five extracellular CAD domains interact
head to head with CAD1 domains from the partner
cells and laterally with other cadherins in a dense
CHAPTER 31 n Intercellular Junctions 551
tangle midway between the two plasma membranes
(see Fig. 30.6).
Desmosomal cadherins connect to cytoplasmic inter
mediate filaments via adapter proteins analogous to
those that connect adherens junction cadherins to actin
filaments. Two proteins related to -catenin, plakoglo-
bin and plakophilin, bind to cytoplasmic domain
of desmosomal cadherins and form a physical link to
desmoplakin, a dimeric protein related to plectin (see
Fig. 35.7). 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).
Although all desmosomes share a common plan, selec
tive expression of isoforms of their component proteins
give desmosomes unique molecular compositions in
various cells. For example, in epidermis, desmoglein-1
and desmocollin-1 are found only in the upper layers,
whereas desmoglein-3 is in the basal layers. This explains
the pathology in autoimmune blistering diseases. Patients
with pemphigus foliaceus make antibodies that react
with desmoglein-1 and disrupt desmosomes in the upper
layers of the epidermis, whereas patients with pemphi-
gus vulgaris produce autoantibodies to desmoglein-3
that disrupt the basal layers. These antibodies are directly
responsible for the disease; transfusion of human auto
antibodies into a mouse reproduces the disease. Other
organs are spared, owing to the restricted expression of
these two isoforms. Mutations in the corresponding
desmoglein genes in mice compromise desmosomes and
cause skin blisters similar to pemphigus.
The development of animal tissues depends on
desmosomes. Loss-of-function mutations can lead to
mechanical failures. For example, mutations in the plako
globin gene can be lethal in mice and humans during
embryogenesis, owing to disruption of the heart. Simi
larly, mutations in the desmoplakin gene cause skin and
cardiac defects that can be fatal. Desmosomal proteins
also participate in signal transduction. For example,
plakoglobin and desmoglein suppress proliferation and
promote differentiation by competing with -catenin
mitogen-activated protein (MAP) kinase pathway. Loss of
desmosomes is associated with the spread of epithelial
cancer cells.
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
552 SECTION VIII n Cellular Adhesion and the Extracellular Matrix

contacts and related assemblies, transmembrane integ
rins 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 resem
blance of hemidesmosomes to half of a conventional
desmosome belies the fact that they are fundamentally
different at the molecular level (Fig. 31.8C). Like desmo
somes, hemidesmosomes have a dense plaque on the
cytoplasmic surface of the plasma membrane that
anchors loops of intermediate filaments. The similarity
ends there.
The hemidesmosomes of simple epithelia use 64
integrin to adhere to laminin-5 in the basal lamina.
Plectin (see Fig. 35.7) links the large cytoplasmic domain
of 4-integrin to keratin intermediate filaments.
More complex hemidesmosomes of stratified epithe
lial cells have, in addition to 64-integrin, a second
transmembrane adhesion protein, type XVII collagen.
The type XVII collagen trimer forms an extracellular
collagen triple helix (see Fig. 29.1) that anchors the

A. Adherens junction B. Desmosome C. Hemidesmosome

Tight
junction

Adherens
junction

2 nm/mm 4 nm/mm 2 nm/mm

CELL 1 CELL 2 CELL 1 CELL 2 IF

IF

Desmoplakin BP230
130 nm
-catenin
BP180
Plectin

2 nm/mm
6 4
Integrin
Actin Plakoglobin
BPAG2
E-cadherin Laminin

BASAL LAMINA

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.
-Catenin and -catenin link the cytoplasmic domain of E-cadherin to actin filaments. B, Desmosome. Two types of cadherinsdesmoglein and
desmocollinlink 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 plako-
globin (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 64 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 intermedi-
ate filaments. (AB, 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;112:535548. C, Micrograph courtesy Jonathan
Jones, Northwestern University, Chicago, IL.)

membrane to the basal lamina. In a blistering skin disease
called bullous pemphigoid, autoantibodies attack
type XVII collagen, so the protein is also called bullous
pemphigoid antigen-2, or BPAG2. This clinical observa
tion and genetic deletions established that both 64-
integrin and type XVII collagen are required for assembly
of stable hemidesmosomes in skin. BPAG1 (bullous
pemphigoid antigen-1; also called BP230) is related to
plectin and helps connect the integrin to intermediate
filaments.
Mutations in the genes for any of the hemidesmosome
proteins cause blistering skin diseases known as epider
molysis bullosa. Pathology can also occur in other tissues
that depend on hemidesmosomes, including the cornea,
gastrointestinal tract, and muscles. Mutations in keratin
genes also cause epidermolysis bullosa (see Fig. 35.6).
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