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Integrating Cells into Tissues

I. Cell-Cell Adhesion.
II. Cell-Matrix Adhesion.
III.Components of the extracellular Matrix.
Yi-Ping Hsueh
Dec 25, 2006

Trans-Well system
Epithelial cells

1. Epithelial cells
2. Cell migration

Cell junctions: Epithelial cells

Cell junctions link to cytoskeletons Tight Junctions

Tight junctions
and adherens
junctions are
linked to the
Freeze-fracture EM
cytoskeleton, and
desmosomes and
are linked to
filaments. a major constituent
of TJ strands

Claudins with molecular masses of ~23 kD comprise a multigene family consisting of

24 members in mice/humans.

The composition of tight junctions. Assembly of epithelial tight junctions and regulation of
proliferation and differentiation.
aPKC, atypical protein
kinase C; huASH1, human ZO-1:
absent, small or homeotic
discs 1; JAMs, junctional
adhesion molecules; MAGI, •Tight junction associated
Membrane-associated MAGUK
guanylate kinase inverted;
MUPP1, multi-PDZ domain
protein 1; Pals1, Protein •In confluent epithelial
associated with Lin-7 1; culture, ZO-1 is restricted
PAR, Partitioning
defective; PATJ, Pals1-
at the junction. In
associated tight junction subconfluent culture, ZO1
protein; PP2A, protein is present in the nuclei.
phosphatase 2A; PTEN,
phosphatase and tensin
homologue; ZO, zonula •ZO-1 interacts with a Y-
occludens; ZONAB, ZO-1- box transcription factor
associated nucleic-acid ZONAB and regulates
EGFR member ErbB2,
which is critical for
regulation of epithelial cell
Some of the components that are thought to become recruited to TJs do not localize proliferation.
exclusively to TJs, but can also localize to the nucleus (for example, huASH1 and ZONAB) or
to other areas of the plasma membrane (for example, PTEN, PP2A and heterotrimeric G PNAS 93:10779 (1996)
proteins). In addition, many TJ components interact directly or indirectly with actin filaments. JCB 160:423 (2003)

claudin-11/oligodendrocyte-specific protein (OSP) claudin-19

* found in CNS myelin and testis * expressed in large amounts in the PNS, not in the CNS
* Tight junction strands in CNS myelin and sertoli cell are absent in * constituted the TJ-like structures of Schwann cells
Osp/claudin-11 null mice * claudin-19–deficient mice lack TJs in Schwann cells and exhibited
* CNS nerve conduction is slowed, hindlimb weakness is conspicuous, and behavioral abnormalities, including defects in locomotion activity.
males are sterile.
Optic nerve

JCB 169, 527 (2005)

Cell 99:649 (1999).

Tight junctions:

1. To seal off body cavities

J. Cell Biol. 169: 527 (2005).
2. To restrict diffusion of membrane components.
Outer mesaxon
3. To maintain blood-brain barrier.
4. To ensure the function of myelin sheath.
Intestinal lumen

Epithelial cells
Inner mesaxon

Basal lamina

Gap Junctions:

1. Allow small molecules (< 1.2 nm in diameter or 1200

Dalton) to pass between adjacent cells, e.g. amino
acids, nucleoside phosphates, cAMP and ions…
2. The channel close in the presence of very high
calcium in the cytosol.

Connexins, connexons, intercellular channels and gap junctions.
* 21 human genes and 20 mouse genes for connexins have been

* Each connexin shows tissue- or cell-type-specific expression, and

most organs and many cell types express more than one connexin.

* Cell coupling via gap junctions is dependent on the specific pattern

of connexin gene expression.

* Regulation of connexin expression is mainly controlled at

transcriptional level.

A diagram showing the relationships between the connexin monomer, the

hexameric assembly of connexins into a connexon and the intercellular joining of
two connexons to form a dodecameric intercellular channel. Clusters of
intercellular channels are known as gap junctions owing to the minute extracellular
'gap' that separates the apposed plasma membranes.

Charcot-Marie-Tooth Disease: Coupling of SCN neurons via gap junctions is important for
the precision of circadian behavior. (NN 8:61, 2004)
1. An X-linked demyelinating disorder of peripheral nervous system.
Individual SCN neurons
2. Caused by mutation in connexin32 (Cx32), which is expressed contain the molecular
machinery necessary to
abundantly in Schwann cells. generate circadian
oscillations. One gap in our
3. Cx32 is concentrated in special regions of Schwann cells. knowledge is the lack of
understanding of how these
4. The gap junctions in Schwann cells allow rapid transportation of single-cell oscillators are
coupled. The new study
the materials from the cell body to periaxonal region. demonstrates that SCN
neurons are coupled through
direct electrical connections.
This coupling is lost in mice
deficient in Cx36. Bottom,
schematics of wheel-running
activity records from WT
and Cx36-deficient mice.
Animals maintained in
constant darkness show
rhythms driven by the
endogenous timing system.
Each horizontal row
represents the activity record for a 24-hour day. Successive days are plotted from top to bottom. The
colored bars represent activity. The WT mice express robust circadian rhythms of locomotor activity with
period shorter then 24 h. The onset of activity is typically under precise control. In contrast, the Cx36-
deficient mice showed rhythms that were weaker and less coherent than controls. Without the Cx36, the
circadian clock still keeps time but lacks the temporal precision that typically characterizes the behavioral

Adherens Junctions and Desmosomes:
Cell junctions: Epithelial cells Cadherin-containing junctions

α- and β-

Cadherin family

Cadherin family: Cadherin

1. E-, P-, N-cadherins…..(more than 300 family
2. Defined by cadherin domain

3. Ca2+-dependent cell adhesion molecules

4. Homophilic interaction.
5. particularly important during early

The dual role of ß-catenin in cell adhesion and transcriptional activation.
Adherens Junctions

* Defects in desmosome-mediated cell–cell adhesion can lead to tissue
fragility syndromes.

* The two organs that appear most vulnerable to these defects are the skin
and the heart.

* Desmosomal proteins are also required for normal embryonic development.

Knockout mice for several desmosomal proteins die in utero. Depending on the
protein studied, death occurs either around the time of implantation, at mid-
gestation or shortly before birth. So far, it appears that structural defects
leading to abnormal histo-architecture and tissue fragility are the main cause
of death.

* There is no evidence that loss of a desmosomal protein would abort specific

cell lineages or differentiation programs.

Eur J Cell Biol. 2005 84:215-23.

Cell junctions: Epithelial cells II. Cell-matrix adhesion

Integrin-containing junctions connect cells to the substratum.

Model for integrin activation

Cell-cell or
cell-matrix interaction

(T-APC interaction)

αvβ3 integrin
Inactive form

Adhesive interactions of fibroblasts Major families of cell-adhesion molecules (CAMs)
integrins and adhesion receptors.

on the flat surface on a 3D matrix of ECM

N-CAMs Major families of cell-adhesion molecules (CAMs)

(nerve-cell adhesion molecule): and adhesion receptors.
1. Calcium-independent homophilic
2. Ig superfamily
3. Particularly important in nervous system.
4. Encoded by a single gene.
5. Important for development.
6. Adhesion of cultured neurons is
inhibited by addition of N-CAMs

Sequence of cell-cell interactions leading to tight binding
of leukocytes to activated endothelial cells and III. Components of the extracellular Matrix
subsequent extravasation
1. Collagen
2. Laminin
3. Fibronectin
4. Proteoglycans

(platelet-activating factor)

III. Components of the extracellular Matrix Collagen fibrils form by lateral interactions of triple helices.
1. Collagen
• The basic structural unit of collagen is a triple helix, which
consists of three coiled subunits (two α1(I) and one α2(I)).
• Repeating motif Gly-Pro-X in collagen proteins.

The side-by-side interactions of collagen helices are
stabilized by an aldol cross-link between two lysine side

Assembly of
collagen fibers
begins in the ER
and is completed
outside the cell.

Structure and
Sheet-forming type IV collagen is a major structural
assembly of
component in basal lamina.
type IV collagen


Major components of the basal lamina Osteogenesis imperfecta (Brittle-bone disease)

Autosomal dominant, lethal disease resulting in death in utero or shortly

after birth
Mutation of collagen: no glycine or hydroxyproline

Both laminin and collagen bind to specific integrins.

3. Fibronectin
2. Laminin Attaching cells to all matrices that contain the fibrous
A large heterotrimeric multiadhesive matrix protein found in all
basal lamina collagens (type I, II, III and V)

4. Proteoglycans: Glycosaminoglycan N-acetylglucosamine or N-acetylgalactosamine

a. Extracellular matrix proteoglycans 1. Heparan Uronic acid

perlecan sulfate
2. Chondroitin
b. Cell surface proteoglycans 3. Keratan
4. Hyaluronan

* Polysaccharide chains in proteoglycans are long repeating

linear polymers of specific disaccharide called
glycosaminoglycans (GAGs).
Linking sugars

Glycosaminoglycan structure Aggrecan
1. Glycosaminoglycans are unbranched, One of the most important extracellular proteoglycans
often high-molecular-weight, sugar
chains that consist mostly of
repeating disaccharide units.
2. Heparan, chondroitin and dermatan
sulphates are normally O-linked
through core-protein serine residues
that are amino-terminal to glycine
and are flanked by sequences rich in
acidic, and poor in basic, residues.
3. The repeating disaccharides of these
glycosaminoglycans are linked
through the sequence xylose–

Syndecan family:
1. Transmembrane heparan sulfate proteoglycans
2. Syndecan-1, -2, -3 and –4.
3. Developmentally regulated.

RBC is 10000 nm in diameter.

Syndecans act as co-receptors of growth and Protein Phosphorylation Regulates the
differentiation factors. Function of Syndecans.

EphB2 phosphorylates syndecan-2 and regulates the

synaptogenesis activity of syndecan-2. PKC phosphorylates syndecan-2 and regulates
left-right asymmetric development.

1. Molecular Cell Biology, fifth edition, by Lodish, Berk, Zipursky,

I. Cell-Cell Adhesion and Communication. 2.

Matsudaira, Baltimore and Darnell. Chapter 6.
J Clin Invest, April 2002, Volume 109, Number 8, 987-991.
3. Nature Reviews Molecular Cell Biology 4; 285-295 (2003)
tight junction
4. Nature Reviews Molecular Cell Biology 4; 926-938 (2003)
adherens junction 5. Nature Reviews Molecular Cell Biology 4; 225-237 (2003)
6. Nature Reviews Molecular Cell Bioloby 1; 91-100(2000)
desmosome 7. Nature Neuroscience 8:61 (2004)
8. J. Cell Biol. 169: 527 (2005).
gap junction
9. Cell 99:649 (1999).
10. JCB 169, 527 (2005)
II. Cell-Matrix Adhesion. 11. Biochimica et Biophysica Acta (BBA) - Biomembranes 1719, 6-23
12. Eur J Cell Biol. 84:215-23 (2005).
focal adhesion (also in non-epithelial cells) 13. PNAS 93:10779 (1996)
14. JCB 160:423 (2003)

III. Components of the extracellular Matrix.

collagen, laminin, fibronectin, proteoglycans