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Trans-Well system
Epithelial cells
1. Epithelial cells
2. Cell migration
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Cell junctions: Epithelial cells
Tight junctions
and adherens
junctions are
linked to the
actin
Freeze-fracture EM
cytoskeleton, and
desmosomes and
hemidesmosomes
are linked to
intermediate
filaments. a major constituent
of TJ strands
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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
binding.
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)
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Tight junctions:
Epithelial cells
Inner mesaxon
Basal lamina
Gap Junctions:
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Connexins, connexons, intercellular channels and gap junctions.
Connexins:
* 21 human genes and 20 mouse genes for connexins have been
identified.
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
output.
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Adherens Junctions and Desmosomes:
Cell junctions: Epithelial cells Cadherin-containing junctions
α- and β-
catenins
plakoglobin
Cadherin family
desmocollin
E-cadherin
desmoglein
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The dual role of ß-catenin in cell adhesion and transcriptional activation.
Adherens Junctions
Desmosome
Desmosome:
* 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.
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Cell junctions: Epithelial cells II. Cell-matrix adhesion
Cell-cell or
cell-matrix interaction
(T-APC interaction)
αvβ3 integrin
Inactive form
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Adhesive interactions of fibroblasts Major families of cell-adhesion molecules (CAMs)
integrins and adhesion receptors.
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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.
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The side-by-side interactions of collagen helices are
stabilized by an aldol cross-link between two lysine side
chains.
Assembly of
collagen fibers
begins in the ER
and is completed
outside the cell.
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Structure and
Sheet-forming type IV collagen is a major structural
assembly of
component in basal lamina.
type IV collagen
Head-to-head
Tail-to-tail
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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)
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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–
galactose–galactose–uronic-acid.
Syndecan family:
1. Transmembrane heparan sulfate proteoglycans
2. Syndecan-1, -2, -3 and –4.
3. Developmentally regulated.
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Syndecans act as co-receptors of growth and Protein Phosphorylation Regulates the
differentiation factors. Function of Syndecans.
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References:
Outline:
1. Molecular Cell Biology, fifth edition, by Lodish, Berk, Zipursky,
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