Intercellular Connections

Learning Objectives

 Cell Adhesion Molecules

 Intercellular Connections.
 Brief description of each type and their
function.
 Cell Adhesion Molecules
(CAMs)
 Important cell surface proteins molecules
promoting cell–cell and cell–matrix
interactions.
 Important for many normal biological
processes
-embryonic cell migration, immune system
functions, wound healing.
 Involved in intracellular signaling pathways
(primarily for cell death/survival, secretion etc.)
 Cell Adhesion Molecules
 (CAMs)
Express 3 major domains:
• The extracellular domain allows one CAM to bind to
another on an adjacent cell.
• The transmembrane domain links the CAM to the
plasma membrane through hydrophobic forces.
• The cytoplasmic domain is directly connected to the
cytoskeleton by linker proteins.
 Cell Adhesion Molecules
(CAMs)
 Interactions between CAMs can be mediated by
:

Binding of an
An adhesion molecule
adhesion molecule on The linker molecule in
on one cell type binds
one cell to the same most cases is
to a different type of
adhesion molecule on Laminin, a family of
cell adhesion molecule
a second cell large cross shaped
on a second cell
Cadherin - cadherin molecules with
Selectins – mucins
multiple receptor
domains.
 CAMs PLAY ROLE IN

• Cell-cell interactions.
• Embryogenesis.
• Immunity(migration of immune cells to the
inflamation center).
• Cell-tissue-organ development.
• wound healing.
• Cancer metastasis.
 Principal classes of cell-adhesion
molecules
 Identified by using specific monoclonal
antibodies (mAbs).
 genes encoding these molecules has shown that
they are structurally different from each other.
 These cell adhesion molecules can be divided
into 4 major families
• The cadherin superfamily
• The selectins
• The immunoglobulin superfamily and
• The integrins
 CADHERINS
• The primary CAMs in adherens junctions and
desmosomes belong to the cadherin family.

• In vertebrates, >100 family members have been

identified with diverse protein structures,but all with
characteristic extracellular cadherin repeats,can be
grouped into classical and non classical cadherins.

• The diversity of cadherins arises from the presence of

multiple cadherin genes and alternative RNA
splicing.
• The role of cadherins is not limited to mechanical adhesion
between cells. Rather, cadherin function extends to
multiple aspects of tissue morphogenesis,including cell
recognition and sorting, boundary formation and
maintenance, coordinated cell movements and the
induction and maintenance of structural and functional cell
and tissue polarity.

• Cadherins have been implicated in the formation and

maintenance of diverse tissues and organs ranging from
polarization of simple epithelia to mechanically linking hair
cells in the cochlea to providing an adhesion code for
neural circuit formation during wiring of the brain.
 CLASSICAL CADHERINS
• The "classical'' cadherins include E-,N-, and P-cadherins,
named for the type of tissues in which they were initially
identified (epithelial, neural, and placental).

• The adhesiveness of cadherins depends on the presence of

extracellular Ca2+, the property that gave rise to their
name (calcium adhering).

• Each classical cadherin contains a single transmembrane

domain, a relatively short C-terminal cytosolic domain, and
five extracellular "cadherin" domains (EC).
If the calcium concentration is less the extra
cellular domain becomes floppy and rapidly
degraded by proteolytic enzymes
 The Cadherin
superfamily

 E-cadherin is thought to be important during

embryonic development, and is also involved in
generating and maintaining epithelial layers in
adult tissues.
 The loss of E cadherin expression has been
linked to the invasive behavior of tumour cells
 The Selectins
 Involved in heterophilic cell-cell interactions.
 Family of Ca+2 dependent carbohydrate
binding proteins, mediate the initial attachment
of leukocytes to the endothelium on the blood
vessel wall during the rolling step of leukocyte
extravasation in inflammation.
 Selectins recognize fucosylated carbohydrate
ligands, especially structures containing Sialyl-
Lewis x (sLex) and Sialyl-Lewis a (sLea),
which are heavily expressed on neutrophils
and monocytes
 The Selectins
 Structural features of selectins
include:
• NH2-terminal C-type Ca2+
dependent lectin like binding
domain, which determines the
ability of each selectin to bind to
specific carbohydrate lingands.
• an epidermal growth factor-like
region.
• a number of repeat sequences.
• a membrane-spanning region
and
• a short cytoplasmic region
 The Selectins
 Selectin family
• Leukocyte-expressed L-selectin(CD62L)
• Endothelial-expressed E-selectin(CD62E)
• P-selectin(CD62P) which is expressed by both platelets and
endothelial cells
 The Selectins

• Recently elevated levels of L-selectin have been observed in the

serum of patients with AIDS and leukemia (1)
• E selectin has been found to regulate adhesion of human
colon cancer cells to the endothelium by binding to sLea and
sLex carbohydrate ligands (2)
 Immunoglobulin Superfamily
Molecules
 Have a series of globular Ig-like
domains, formed by disulfide bonds.
 Mediate Ca-independent cell
adhesion.
 Primarily homophilic cell-cell adhesion but also
some heterophilic.
 Activate intracellular signaling pathways.
 Play critical role during morphogenesis and
differentiation of muscle, glial and nerve cells
 In neurons promote the formation of myelin
 In vascular endothelial cells leukocyte adhesion
and extravasation.
 Immunoglobulin Superfamily
Molecules
 Consists of more than 25 molecules.
 Important ones being:
• Intracellular adhesion molecule 1(ICAM1; CD54)
• Intercellular adhesion molecule 2 (ICAM2),
• Vascular cell adhesion molecule1 (VCAM1; CD106),
• Platelet endothelial cell adhesion molecule 1 (PECAM
1; CD31) and
• the mucosal addressin cell adhesion molecule 1
(MAdCAM1).
 ICAMs
• ICAM-1 (CD54), ICAM-2 (CD102) are counter receptors
for leukocyte β2 integrins
• ICAM-1 is expressed on leukocytes, fibroblasts,
epithelial cells & endothelial cells
• ICAM-2 has similar distribution but is not regulated by
cytokines as ICAM-1
VCAMs
• VCAM-1 (CD106) is expressed on surface of activated
endothelium, dendritic cells, tissue macrophages &
bone marrow fibroblasts
• It interacts with leukocyte integrin α4 β1 on
eosinophils, monocytes & with α4 β7 on activated
peripheral T-cells
PECAMs
• PECAM-1 (CD31 or endoCAM) is found on
endothelial cells, on platelets, some monocytes &
neutrophils
• Is involved in homophilic adhesion

NCAMs
• Is expressed on most of the nerve cells
• Play an important role in fine tuning of
adhesive interactions during development &
regeneration
“Although cadherins & Ig family members are frequently
expressed on the same cells, the adhesions mediated by
cadherins are much stronger & are responsible for
holding cells together, segregating cell collections into
 The integrins
 Cell adhesion receptors responsible for the cell
extracellular matrix adhesion
 Important signal transduction receptors for
regulation of cell growth
 Present in membranes of all cells except
erythrocytes.
 Composed of heterodimers consisting of two
non-covalently associated subunits,α and β, both
of which are necessary for adhesive binding.
 The integrins
 Fifteen different α and eight
different β subunits give rise to over
twently different heterodimeric
combinations at cell surfaces.
 Bind epithelial and muscle cells to
laminin in the basal lamina
 Allow platelets to stick to exposed
collagen in a damaged blood vessel
 Allow fibroblasts and white blood
cells to adhere to fibronectin and
collagen as they move
• Integrins can bind to extracellular matrix (ECM)
glycoproteins including collagens, fibronectins,
laminins,and cellular receptors such as vascular cell
adhesion molecule-1 (VCAM-1) and the intercellular
cell adhesion molecule (ICAM) family.

• Integrins also play key roles in the assembly of the

actin cytoskeleton as well as in modulating signal
transduction pathways that control biological and
cellular functions including cell adhesion, migration,
proliferation, cell differentiation,and apoptosis.
 FUNCTION OF INTEGRINS

1. Attachment of cell to ECM

2. Signal transduction from ECM to cell
have relation to-
-cell growth
-cell division
-cell survival
-cellular differentiation
-apoptosis
-cell migration during
embryogenesis, thrombosis,
haemostasis,wound healing etc.
Leukocyte migration and CAM
 CAMs in leukocyte adhesion deficiency

• Leukocyte adhesion
deficiency (LAD), is a
rare autosomal recessive disorder
characterized
by immunodeficiency resulting in
recurrent infections.

• LAD is currently divided into three

subtypes: LAD1, LAD2, and the
recently described LAD3, also
known as LAD-1/variant.

• In LAD3, the immune defects

are supplemented by a
Glanzmann thrombasthenia-like
bleeding tendency.
 CAMs in CANCER and METASTASIS
• The function of epithelial (E)-cadherin is decreased in most
epithelial tumors during cancer progression. Loss of E-
cadherin function elicits active signals that support tumor-cell
migration, invasion and metastasis.

• Loss of E-cadherin can be accompanied by increased level of

other cadherins such as N-cadherin which promotes tumor
cell motility and migration.

• E cadherin functions as a Tumor Suppressor.

• E cadherin loss enables disaggregation of cancer cells from

one another.
• CDH1 mutations involved with several cancers:
Breast, Liver, Prostate, Stomach, Endometrium,
Ovary, and Lung.

• Loss of E cadherin Function Correlates with Poor

Prognosis.

• Accumulating evidence from several preclinical

models confirms that tumor cell interactions
through selectins and integrins actively
contribute to the metastatic spread of tumor cell.
 CAMs in CARDIOVASCULAR
PATHOLOGY
A.Coronary Artery disease: The infiltration of monocytes
and T- lymphocytes, which initiates the atherosclerotic
process is mediated by adhesion receptors.
There are increased level of β-1 integrin, VCAM-1,
ICAM-1 and E-selectin in patients with atherosclerosis.

B.Thrombosis: Activation of αIIbβ3 receptor can

result in platelet adhesion even if Arachidonic Acid
pathway is blocked.
 CAMs IN INFLAMMATORY BOWEL DISEASE

• VCAM- 1 concentration were higher in patients with

active ulcerative colitis compare to inactive ulcerative
colitis.

• VCAM-1 concentration were also greater in patients

with both active and inactive crohns disease than
controls.

• Patients with active crohns disease had higher

ICAM-1
concentration than control.
 CAMS IN ALZHEIMER DISEASE
• Neuropathologically, AD is primarily characterized
by intraneuronal neurofibrillary tangles (NFTs) of
hyperphosphorylated tau and extracellular
deposits of mainly aggregated Aβ peptide, known
as senile or neuritic plaques.

• The role of CAMs in events considered central to

the pathogenesis and progression of Alzheimer’s
disease (AD) such as amyloid-β (Aβ)
metabolism, neuronal plasticity, inflammation,
and vascular changes.
 Intercellular
Connections.
OCCLUDING JUNCTIONS
• Tight Junctions (Zona Occludens)
ANCHORING JUNCTIONS
Actin filament attachment sites
• Cell- cell junctions (Zonula Adherens)
• Cell-matrix junction (Focal Adhesions)
Intermediate filament attachment sites
• Cell-cell junction (Desmosomes)
• Cell-matrix junction (Hemidesmosomes)
CHANNEL FORMING JUNCTIONS
• Gap junctions
SIGNAL RELAYING JUNCTIONS
• Chemical synapse
 Tight Junctions
 Also known as Zona Occludens.
 Surround the apical margins of the cells in
epithelia such as the intestinal mucosa, the walls of
the renal tubules, and the choroid plexus.
 Made up of ridges—half from one cell and half
from the other—which adhere so strongly at cell
junctions that they almost obliterate the space
between the cells.
 Permit the passage of some ions and solute in
between adjacent cells (paracellular pathway) and
the degree of this ―leakiness‖ varies, depending in
part on the protein makeup of the tight junction.
 Tight Junctions

 Basic architectural principle - transmembrane

proteins are linked to a cytoplasmic plaque that is
formed by a network of scaffolding and adaptor
proteins, signalling components and actin-
binding cytoskeleton
 Tight Junctions
 TRANSMEMBRANE TIGHT JUNCTION PROTEINS:
 Tight Junctions contain two principal types of
Transmembrane protein components – tetraspan
and single-span transmembrane proteins.
 The tetraspan proteins are:
• occludin and the claudins
• have both the N- and C-termini in the cytosol.
• form the paracellular permeability barrier and determine
the capacity and the selectivity of the paracellular
diffusion pathway.
 The single-span transmembrane proteins are the
junctional adhesion molecules (JAMs),
 Functions of Tight Junctions
 Paracellular permeability:
• allow the passive selective diffusion of ions and small
hydrophilic molecules through the paracellular
pathway across epithelia and endothelia.
• the claudin composition of TJs is a major determinant
of the permeability properties of a tissue.
• Occludin regulates the paracellular diffusion of small
hydrophilic molecules, and regulates the
transepithelial migration of neutrophils.
• The passage of solute depends upon its size and
charge.
 Functions of Tight Junctions
 Cell proliferation, polarity and differentiation:
• Several studies have linked TJs to the regulation of
cell proliferation and cell polarity.
• Occludin suppresses oncogenic Raf-1 signalling
(Wang et al., 2005) and interacts with
ZONAB, thereby regulating gene expression, cell
proliferation and epithelial morphogenesis (Matter
and Balda, 2007; Sourisseau et al.,2006)
• Occludin has also been linked to the regulation of
various subcellular signalling pathways, such as MAP-
kinase-dependent pathways.
Disease of Tight Junctions
 Gap Junction

 Gap junctions are clusters of intercellular

channels that allow direct diffusion of ions and
small molecules between adjacent cells.
 At gap junctions, the intercellular space narrows
from 25 nm to 3 nm.
 gap junctions were first discovered in
myocardium and nerve because of their
properties of electrical transmission between
adjacent cells (Weidmann 1952; Furshpan and
Potter 1957).
 Gap Junction

 The intercellular channels are formed by head-to-head

docking of hexameric assemblies (connexons) of
tetraspan integral membrane proteins, the connexins
(Cx) (Goodenough et al. 1996).
 Gap Junction

 Electron microscopy of gap junctions joining adjacent hepatocytes

in the mouse. The gap junction (GJ) is seen as an area of close
plasma membrane apposition
 Function of Gap Junction
 The diameter of the connexon channel is normally about
2 nm, which permits the passage of ions, sugars, amino
acids, and other solutes with molecular weights up to
about 1000 Dalton.
 Function as suppressors of somatic cell mutations -loss
of a critical metabolic enzyme or ion channel in one cell
compensated by its neighbours.
 Are particularly important in cardiac muscle: the signal
to contract is passed efficiently through gap
junctions, allowing the heart muscle cells to contract in
tandem.
 A gap junction located in neurons referred to as an
electrical synapse are important in neurotransmitter
release
 Disease associated with

Gap Junctions
20 different genes code for connexins in humans, and
mutations in these genes can lead to diseases that
are highly selective in terms of the tissues involved.
 In humans, mutations in Cx32 underlie X-linked
Charcot-Marie-Tooth syndrome, a common peripheral
demyelination neuropathy.
 mutations in Cx47 result in a central demyelinating
condition.
 disorders of the skin and the auditory system
accompany mutations in Cx31 andCx30.
 Familial cataracts are commonly associated
with
mutations in either Cx46 or Cx50.
 Desmosomes
 Also known as macula adherens is a cell structure
specialized for cell-to-cell adhesion.
 Are molecular complexes of cell adhesion proteins and
linking proteins that attach the cell surface adhesion
proteins to intracellular keratin cytoskeletal filaments.
 The cell adhesion proteins of the desmosome, desmoglein
and desmocollin, are members of the cadherin family.
 On the cytoplasmic side of the plasma membrane, there
are two dense structures called the Outer Dense Plaque
(ODP) and the Inner Dense Plaque (IDP).
• The Outer Dense Plaque is where the cytoplasmic domains of
the
cadherins attach to desmoplakin via plakoglobin and
plakophillin.
• The Inner Dense Plaque is where desmoplakin attaches to the
Desmosomes
 Hemidesmosomes
 Hemidesmosomes look like half-desmosomes
that attach cells to the underlying basal lamina.
 Rather than using
desmogleins, hemidesmosomes use
desmopenetrin cell adhesion proteins,which are
members of Integrin family.
 The integrin molecule attach to one of many
multi-adhesive proteins such as
laminin, resident within the extracellular
matrix, thereby forming one of many potential
adhesions between cell and matrix.