Cell adhesion, junctional structures,

the epithelial cell

Prof. Dr. Pál Röhlich

Dr. Anna L. Kiss

Department of Anatomy, Histology and Embryology

Semmelweis University
Budapest
2017
 Adhesion
Specific bindings between a.) individual cells
b.) cells and extracellular matrix (ECM).
- Integrates cells in a multicellular organism,
- crucial for organisation of tissues and organs.
- intracellular anchoring to filaments of the cytoskeleton (actin
microfilaments or intermediate filaments).

I. Adhesion between cells

II.Adhesion between cell and extracelluar matrix

 I. Adhesion between cells:
Adhesion proteins in the plasma membrane (integral membrane proteins).

Significance:
• in embryonic development (cell associations, switching between cell
associations, specific attachment between nerve and muscle, formation of
interneuronal connections (synapses), formation of epithelial layers, etc. …)

• in adult orgnisms:
• long lasting connections (e.g. epithelia, nervous system)
• or temporal connections (e.g. in the immune system).
• pathology: metastasis of cancer cells, …
Cell Adhesion Molecules (CAMs)
- are intramembrane (transmembrane) proteins located on the cell surface;

- involved with the binding with other cells or with the extracellular matrix (ECM) in
the process called cell adhesion;

- many weak bonds are formed between the cells.

These proteins are typically transmembrane receptors and are composed of three domains:

a.) an intracellular domain that interacts with the cytoskeleton,

b.) a transmembrane domain,

c.) an extracellular domain that interacts either with other CAMs of the same kind (homophilic or
homotypic binding) or with other CAMs or the extracellular matrix (heterophilic or heterotypic
binding).
Cell adhesion molecules

homotypic binding

heterotypic binding
 Cell adhesion molecules

Ca2+-dependent:

cadherins
selectins
integrins

Ca2+-independent:

IgG-like- CAMs (immunoglobulin superfamily)

 Cell-cell adhesion molecules

Ca2+-dependent:
1. Cadherins: Adhesion proteins with a single transmembrane portion.
The extracellular part usually consists of 5 repeating domains. Extracellular tip
of the molecule (last domain) binds a similar domain of cadherin in the
opposite membrane (homotypic binding).

Stability of the domains is Ca2+-dependent (without Ca2+ binding, the

extracellular part of the molecule collapses and the cells become dissociated).

Cadherins form a relatively strong binding between cells.

Cadherin-superfamily (about 180 cadherin types in human, „variations on a theme”), Nomination
according to cell type and characters, recently numbers.

Classical cadherins: E-cadherin (epithelial, in cells of epithelium and morula),

P-cadherin (placentar, in heart, lung, gut),
N-cadherin (neural, in development of the nervous system,
synaptogenesis), M-cadherin (muscular, myoblasts),
R-cadherin (retinal, outgrowth of nerves),
VE-cadherin (vascular endothelial),
K-cadherin (kidney), …

In a single cell several cadherin types can occur, expression of cadherins.

Appearance of E-cadherin in
the morula stage of embryonic
development. Mouse. Cells of
the morula closely adhere to
one another: compaction.

1.5 days 3.5 days

 2. Selectins

Ca2+- dependent, weak heterotypic bindings.

The terminal domain has binding site for certain membrane glycoproteins of other
cells.
Role: recirculation of lymphocytes in the organism, adhesion of leucocyte to the
endothelial surface of blood vessels. („rolling”), …
L-Selectin (on leucocytes), P-selectin (on blood platelets), E-selectin (on
endothelium)

anchoring proteins
actin filaments
3. Integrins:
Ca2+-dependent, heterotypic binding, α and β chains,
•binding site on the extracellular end.
•24 different integrins. Large majority binds to components of the
extracellular matrix, but a few of them to cells.
These are:
LFA-1 (αLβ2) Integrin (on leucocytes), binds I-CAM and V-CAM
adhesion molecules on endothelial cells. Migration of leucocytes
through the wall of the blood vessel.
Binding site

Mac-1 (αMβ2) Integrin (on macrophages and lymphocytes)

α-chain β-chain
 Ca2+-independent

Immunoglobulin-like adhesion molecules: CAMs

No Ca-dependence!
Immunoglobulin-domains. Weak adhesion, fine regulation. Disulfide-bond

N-CAM (neural cell adhesion molecule). Homotypic bindings.

Negative charges on the sialic acid molecules can modulate binding.
Role: in development of the nervous system, formation of ganglia,
outgrowth of nerve fibers.
I-CAM, V-CAM (in the lining cells of blood vessels). Heterotypic
bindings with integrins of leucocytes. L1-CAM (cell migration,
differentiation), PECAM (adhesion between blood platelets and
endothelium).
 II. Adhesion between cells and
extracellular matrix (ECM)
Strong connection between cells and ECM
ECM: produced by the cells: 1.) fibers; 2.) proteoglycans, glucosaminoglycans; 3.)
matrix binding glycoproteins
1.) Connective tissue fibers:
Collagen fibers:
Large family of collagen proteins. Most frequently occur:

collagen type I: composed of fibrous collagen molecules,

resistant against pull, thin collagen fibrils form
bundles of various thicknesses: thick collagen fibers.

collagen type III: can form thin fibers

often interconnected with each other (reticular fibers).
2.) Glycosaminoglycans, proteoglycans.

Glycosaminoglycans (GAGs) are long sugar chains, composed of disaccharide

units and carrying acidic groups (carboxyl, sulfate), therefore they contain many
negative charges. High water-binding capacity!
Proteoglycans (PGs): consist of a core polypeptide chain and of GAG chains
laterally bound with covalent bonds.
3.) Matrix-binding glycoproteins:

heterogenous proteins many binding sites for ECM components and

cells. They form bridges in between them. The most promeinent matrix-
binding proteins: laminin és a fibronectin.
 II. Adhesion between cells and
extracellular matrix (ECM): integrins
Functions of integrins:
:
• Adhesion between cell and ECM (Attachment of the
cell onto the underlying structure is important!)

• Local integration of the cytoskeleton with the ECM

• Signal transduction from inside outward and reversed

Indirect significance: cell migration, cell division, development, differenciation

Integrin as signal transduction
molecule
Active and inactive forms. Binding of a ligand to the integrin molecule on
one side of the membrane activates the molecule on the other side of the
membrane (transduction of a signal across the membrane).

The role of talin. Activation of signal transduction pathways (binding to the

β-chain activates extracellular part of the molecule which in turn strongly
binds ECM components.)

ligand binding

talin binding
Integrins play important role in cell
migration
Integrin internalistion: via
caveolae

Recycling/or lysosomal
degradation
Integrins in signal transduction
 The epithelial cell
Surface epithelium: Epithelial cells are attached to each other laterally and form a continuous layer on
free surfaces. General characteristics of the epithelial cell are introduced on the example of the columnar
epithelium.
The epithelial cell is a polarized cell: cell organelles, cytoskeletal constituents, membrane domains and cell
junctions are arranged in a characteristic pattern and orientation.

Apical (luminal) surface


apical surface

Golgi
gap junction

centrosome lateral surface

 Intestinal
epithelium

basal lamina basal surface

 Columnar epithelial cell showing localisation

of cell junctions and surfaces.
 Epithelial cells
Poligonal cells

apical

lateral

basal
Apical surface
• glykocalyx
• microvilli (brush border)
• stereocilium
• cilia, flagellum

Function:
– protection (mechanical, biological)
– diffusion barrier
– absorption,
– movements
– secretion (exocytosis)
– transcytosis
 Glycocalyx

Function
cell-cell recognition
(MHC, blood groups)
enzyme-function (intestine: enterocytes)

cell membrane
 Microvilli (brush border)

Microfilaments Glykocalyx

0.08 m

1 m

Terminal web – bundles of actin

Microvilli (brush border)
To increase the surface
Stereocilia
length: 8-10 µm
diameter: 1 µm
actin filaments

stereocilia

Ductus epidydymis
Cilia
length: 5-10 µm
diameter: 0,2 µm

Function: movements
Structure: microtubules (tubulin)
nexin (connects microtubules)
dynein (outer and inner)
Cilia (9x2 +2) and basal body (9x3)
 Basal surface
Basement membrane: inhibits the free diffusion
induces polarity
regeneration
filter (kidney)

epithelial
cell

• lamina basalis

lamina lucida (rara)

lamina densa

• lamina fibro-reticularis
 Basal surface
Basal or basement membrane (light
microscope)
– Basal lamina:
Type IV-collagen + adhesion molecules
Glycoproteins (laminin, fibronectin) +
proteoglycans
• Lamina rara externa (lamina lucida)
• Lamina densa
• Lamina rara interna
– Lamina fibroreticularis:
TypeIII-collagen
 Basement membrane
A thin layer (40-120 nm) under the basal surface
of the epithelium, only seen with the electron
microscope

lamina rara

lamina densa

Electron microscopy

lamina fibroreticularis
 Molecular structure of the basal lamina.
Components:

Collagen type IV (these fibrous protein molecules form a multilayered network: lamina densa, a
resistant and plastic layer)
Laminin (matrix-binding molecule)
Perlecan (proteoglycan)
Nidogen (a small protein with multiple binding sites)
Fibronectin (matrix-binding molecule)

Binding to the plasma membrane by: integrin (laminin-receptor), dystroglycan and syndecan
(membrane proteoglycans)
Integrin Perlecan Dystroglycan

Laminin

Nidogen

collagen Type IV
lamina densa

Fibronectin
 Biological significance of the basal lamina:

1. mechanical: a.) binds the epithelial layer firmly to the ECM. Important e.g. in the strong
binding of the epidermis to the underlying connective tissue.
The basal lamina is a continuous layer, b.) inhibits immigration of connective tissue cells into
the epithelium or free emigration of epithelial cells into the connective tissue (cancer!).

2. cell biological: the basal lamina sends information signals through integrins into the cell
interior and is therefore important in survival and division of the epithelial cell as well as in the
maintenance of cell polarity. During epithelial regeneration it „leads” the epithelial cells to
cell-free areas. Without basal lamina no continuous epithelial layer can be formed.
Genetic defects of basal lamina proteins are lethal for the embryo in an early developmental
stage.

3. molecular sieve: filters blood from the capillaries of the renal glomerulus (ultrafiltrate) and
retains proteins and cells in the blood. In genetic defects of one of its molecular components
blood or blood proteins appear in the urine. The pore size of the filter depends primarily on
proteoglycans in the basal lamina.
 Basal striation

membrane invaginations+mitochondria
 Lateral surface: cell junctions
• Mechanical connecting structures:
– macula adherens (desmosoma)
– zonula adherens
– hemidesmosoma

• Diffusion barriers
– zonula occludens (tight junction)

• Communicating junctions
– nexus (gap junction)

Long-lasting coupling between cells and between cell and ECM with various
functions.
General structure: adhesion molecules,
adaptor proteins,
cytoskeleton
Desmosomes and zonula adherens

adaptor proteins:
(cateins, vinculin, α-
actinin)
adaptor proteins

integral membrane
protein, adhesion mol.
Desmosomes
Zonula adherens, desmosomes
Hemidesmosome (half-desmosome):
Adhesion molecules: integrins (e.g. laminin receptor α6β4)
Adaptor molecules: plectin, dystonin
Cytoskeleton: intermediate filaments (e.g. keratin in epithelium)
ECM: laminin (lamina basalis)
 Hemidesmosomes
• It makes stronger the connection of the epithelial cells to the lamina
basalis

integrins
Hemidesmosomes
Zonula occludens (tight junction)

Integral membrane protein: occludin, claudin

adapter molecules: ZO1, ZO2

cytoskeleton: microfilaments (actin)í

Zonula occludens (tight junction)

ZO1, ZO2 protein

 Functions of tight junction

• Diffusion barrier in the plasma membrane (blocking lateral diffusion in the membrane), the belt-
like tight junction divides the plasma membrane into an apical and baso-lateral domain (with
different sets of molecules)

• Diffusion barrier in the intercellular space: free diffusion of substances between neighboring
cells is blocked.
•
Biological significance: controlled and unidirectional transport across the epithelial layer
apical membrane

baso-lateral membrane

baso-lateral membrane
 Zonula occludens (tight junction)
Cell junction functioning as a diffusion barrier in the intercellular space and in the plasma
membrane.
Localisation: belt-like, running circumferentially close to the luminal surface.



 Nexus (gap junction)
Patch-like contacts between two cells: large number of channels (connexons) in the
opposite membranes, that are bound to each other in the intercellular space to form
continuous channels between the two cells.

cell membranes

intercellular space

connexon (connexin subunits)

2 connexons
Molecular structure:
opposite plasma
membranes
connexon: a complex composed of 6 transmembrane proteins
(connexins), that surround a central canal. A similar complex in the
opposite membrane is joined to it in the intercellular space. A
continuous canal is formed which leads from one cell into the other
making communication by free diffusion of low molecular weight
substances possible.
Connexin: Transmembrane proteins
Gap junction: patch-like membrane domain with densely packed
connexons
Diversity of connexons, in different cell types. Combinations of
different connexins in a connexon.
Examples of connexin isoforms:

Cx50 in the crystalline lens of th eye, defects lead to
glaucoma
Cx26 in sensory cells of the inner ear, defects cause
auditory malfunction
Cx32 in nerve fibers, defects lead to problems in nerve
conduction.
 Nexus
Biological significance:
Communication between cells: nutrient transport (cells of the lens, osteocytes,
follicular cells surrounding the oocyte in the ovary), synchronized reaction for
signals during the development etc.
Electrical connection between cells: electrical synapsis: ions can migrate
between cells, stimulus can be transmitted without delay;
In heart muscle: synchronized contraction
Nexus (gap junction, macula communicans)

gap
junction

 
Interdigitation
Cell junctions in polarized
cells

Hemidesmosom
es
 Textbook: Essential cell biology, 3rd ed. p. 689-690, 693-707,

+ szövettan + lecture summarized in the present ppt presentation

Sources of figures:

 Röhlich: Szövettan, 3. edition, Semmelweis, Budapest

 Alberts – Johnson – Lewis – Raff – Roberts – Walter: Molecular

biology of the cell. 5. edition, Garland Science

 Specimens and/or micrographs, or drawings of P. Röhlich

 Junqueira – Carneiro: Histologie, 6. edition, Springer