ANATOMY OF DERMO-

EPIDERMAL JUNCTION AND

ITS APPLIED ASPECT

Presenter- Dr. Siddhartha Dash

Moderator- Dr. Liza Mohapatra
 ULTRASTRUCTURE OF SKIN

 Consists of-
1.EPIDERMIS : Divided into-
a. Stratum Basale
b. Stratum Spinosum
c. Stratum Granulosum
d. Stratum Corneum
• Stratum basale & stratum spinosum together are
called Stratum Malphigi.
• In palms & soles, an additional Stratum
Lucidum is present.
• Keratinocytes constitues 95% of epidermal cells.
 2. DERMO-EPIDERMAL JUNCTION.
3. DERMIS : Organised into-
a. Papillary Dermis
b. Periadnexal Dermis
c. Reticular dermis
• Papillary & periadnexal dermis together are
called Adventitial dermis.
4. HYPODERMIS/ SUBCUTIS:
Consists of a layer of subcutaneous fat, which is
separated from rest of the body by a vestigial
layer of striated muscle.
 DERMOEPIDERMAL JUNCTION
 Interface between the lower part of epidermis
and the top layer of dermis.
 Consists of :

a. Basal Keratinocytes
b. Dermo-epidermal basement membrane
zone (BMZ).
 BMZ consists of a number of extracellular
matrix macromolecules, most of which are
glycoproteins (stained by PAS).
BASEMENT MEMBRANE ZONE
 Consists of Basal & Fibro-reticular Lamina.
 Ultrastructural examination of the BMZ by
transmission electron microscopy shows the
presence of 2 distinct layers with different optical
densities.
1. Upper electron luscent Lamina Lucida.
2. Lower electron dense Lamina Densa.
 Lamina lucida & Lamina densa together are
called Lamina basalis/ Basal lamina.
 Lamina fibro-reticularis/ Fibro-reticular
lamina : Basal lamina is attached with reticular
lamina through the anchoring fibrils.
ELECTRON MICROSCOPIC VIEW OF DERMO-EPIDERMAL JUNCTION
SCHEMATIC REPRESENTATION
 In order to hold this structure together, there are several
macromolecules, which binds among themselves. They are:
1. Intermediate filament (IF) components
• Keratin 5
• Keratin 14
2. Hemidesmosomal plaque components
• 230 kDa bullous pemphigoid antigen (BP230/BPAG1)
• Plectin
3. Transmembrane components
• α6β4 integrin
• Type XVII collagen (180 kDa bullous pemphigoid
antigen/BPAG2)
• α3β1 integrin
• Type XIII collagen
• Syndecans 1 and 4
4. Lamina lucida/lamina densa components
• Laminin 332 (laminin 5)
• Laminin 311 (laminin 6)
• Laminin 511 (laminin 10)
5. Lamina densa components
• Type IV collagen
• Laminin 111 (laminin 1)
• Nidogen
• BM‐40/SPARC
• Perlecan
6. Anchoring fibril components
• Type VII collagen
• GDA‐J/F3 antigen
 KERATIN INTERMEDIATE FILAMENTS

 Major structural proteins of the epidermis.

 2 types of keratin encoding genes : Basic (1-8) &
Acidic (9-19).
 To be functional, the keratin pairs must form into
heterodimers i.e. One acidic & one basic keratin
monomer. So they’re called obligate
heterodimers.
 Keratin dimers assemble into a network of 10
nm keratin filaments.
 Keratin 5 & 14 are found in basal keratinocytes.
At the lowest level, 2
monomers associate with
each other to form a
twisted dimer.

2 dimers then line up to

form a stagerred
tetramer.
Tetramers then link
end to end to form
one strand of an
intermediate
filament.

Eight strands stack

together & twist
around each other
to form a rope like
keratin filaments of
size 10 nm.
APPLIED ASPECT :-
•Mutations in keratin 5 & 14 cause Epidermolysis
bullosa simplex.
EBS SUBTYPES PATTERN OF TARGETED
INHERITANCE PROTEINS
1. EBS- DOWLING- AD Missense mutations Keratin 5 & 14
MEARA

2. EBS- LOCALIZED AD Missense mutations Keratin 5 & 14

(WEBER- COCKAYNE)

3. EBS-OTHER AD Missense mutations Keratin 5 & 14

GENERALISED

4. EBS-MOTTLED AD Missense mutations Keratin 5

PIGMENTATION

5. EBS-MIGRATORY AD Framshift mutations Keratin 5

CIRCINATE

6. EBS-AR AR Nonsense mutations Keratin 14

 INTEGRINS
 Transmembrane receptors that bind actin
cytoskeleton to the extracellular matrix ( only
exception is α6β4-integrin that binds to keratin
filaments).
 Obligate heterodimers containing one α subunit
& one β subunit.
 18 different α subunit- α1-11, αD, αE, αL, αM,
αV, α2B, αX.
 8 different β subunit- β1-8.
 Attach to laminin molecules in the extracellular
matrix.
 Mutation of α3 integrin forming gene can cause
junctional epidermolysis bullosa.
 COOH
terminal

Laminin

NH2 terminal

Interaction of integrin with actin & laminin

Kindlin 1 protein – Mutation causes Kindler’s Syndrome.
 HEMIDESMOSOMES

 Electron dense attachment complexes which

extend from the intracellular component of basal
keratinocytes to lamina lucida.
 Intracellular domains bind to keratin
intermediate filaments & extracellular domains
binds to laminin 332.
 5 major components : a) Plectin

b) BPAg1e / BP230
c) α6β4 integrin
d) BPAg2 / BP180 / Type-
XVII collagen
e) Tetraspanin CD151
SCHEMATIC REPRESENTATION
 2 types of hemidesmosomes :
a) Type 1 HD :
 Found in stratified & pseudostratified epithelium.

 Contains all five components.

b) Type 2 HD :
 Found in simple epithelium.
 Contains only α6β4 integrin & plectin.
 PLECTIN & BPAG1E
 They differ only in their c-terminal domain where plectin &
BPAg1e consists of 6 & 2 copies of plakin repeat domains
respectively.
 Consists of :
a) C terminal domain : Also called as universal IF- binding
site & binds with keratin 5 & 14.
b) Central rod domain: Mediates self association.
c) Plakin domain : consists of several spectrin repeats.
d) F-actin binding domain / ABD domain : Binds with β4
integrin.
 Mutation of plectin causes epidermolysis bullosa
simplex with muscular dystrophy (EBS-MD).
 Defects in BP 230 gene causes epidermolysis bullosa
simplex &Autoantibodies against it causes bullous
pemphigoid.
 β4 INTEGRIN
 Consists of a larger cytoplasmic domain ( > 1000
residues) which binds with plectin, BP 230 &
BP180.
 Each cytoplasmic domain consists of five
globular domain namely :
a) Membrane proximal Na+ - Ca2+ exchanger
motif.
b) 2 pairs of fibronectin type Ш domain ( Fn Ш
1,2 & Fn Ш 3,4) : Binds with ABD domain of
plectin & BP 230.
c) Connecting segment.
d) C- terminal end.
 α6 INTEGRIN
o Consists of :
a) N- terminal long extracellular domain : Binds
to BP 180, CD151 & laminin 332.
b) Transmembrane domain.
c) Short intracellular domain.
 Mutation of α6β4 integrin causes junctional
epidermolysis bullosa with pyloric atresia
BPAG2 / COLLAGEN ХVІІ :
 Contains :
a) Globular intracellular domain :
• It is approximately 70kDa & interacts with β4 integrin, plectin & BP
230.
b) Transmembrane stretch.
c) Large c-terminal ectodomain :
• It is approximately 120kDa consisting of 15 collagenous subdomains
flanked by short non collagenous stretches.
• Binds with laminin 332 & integrin α6.
• May be important for regulation of keratinocyte detachment from the
basement membrane.
o Mutation causes generalised intermediate junctional epidermolysis
bullosa & Autoantibodies against it causes bullous pemphigoid
 TETRASPANIN / CD151
 It is a cell surface protien belonging to tetraspan
superfamily of transmembrane proteins.
 Consists of 4 transmembrane domains forming a small and
a large extracellular loops with short intracellular N- & C-
terminal tails.
 Interacts with integrin α6 via its extracellular loop.
 Mutation causes Pretibial EB, nephritis, deafness, β
thalassemia minor
ULTRASTRUCTURE OF HEMIDESMOSOME
APPLIED ASPECTS :
 Plectin :-
• Mutation causes epidermolysis bullosa simplex with
muscular dystrophy (EBS-MD).
• Autosomal recessive inheritance.

 α6β4 integrin :-
• Mutation causes junctional epidermolysis bullosa
with pyloric atresia .
• Autosomal recessive inheritance.

 BP230 / BPAg1e :-
 Defects in gene causes epidermolysis bullosa simplex .
 Autoantibodies against it causes bullous pemphigoid.

 BP180 / Collagen XVII :-

• Autoantibodies against it causes bullous pemphigoid.
• Mutation causes generalised intermediate junctional
epidermolysis bullosa.
• Autosomal recessive inheritance.
 CD151 : Pretibial EB, nephritis, deafness, β
thalassemia minor.
 Mucous membrane pemphigoid – Auto-antibodies
against BP230, BP180, α6β4 integrin ,laminin 5
& collagen VII.
 Pemphigoid gestationis – Auto-antibodies against
BP180 & BP230.
 Lichen planus pemphigoides – Auto-antibodies
against BP180 & BP230. A new antigen of 200
kD of keratinocyte derivation has been identified.
 Linear IgA disease – Autoantibodies against
LABD -1 (70 kD) & LAD – 1 (120 kD), cleaved
portion of extracellular domain of BP180.
 SYNDECANS

 Family of transmembrane core proteins capable of

carrying heparan sulfate(HS) & chondroitin
sulfate(CS) chains.
 4 syndecan genes are present in vertebrate i.e.
Syndecan 1, 2, 3 & 4.
 Syndecan 1 is most abundant in epithelial
keratinocytes. It induces epidermal proliferation.
 Consists of 3 domains:
a) short cytoplasmic domain.
b) single span transmembrane domain.
c) extracellular domain with attachment sites for 3-5
HS or CS chains.
 HS chains:
a) interact with growth factors like FGF, VEGF,
TGF-β.
b) interact with various extracellular matrix
proteins such as fibronectin & antithrombin 1.
 Role of CS chains are not clear but a recent
study suggest that they cooperate with HS
chains of syndecan 1 & 4 in binding to the
heparin binding growth factors midkine &
pleiotrophin & to the ECM protein laminin.
 Expession of syndecans are increased during
wound healing.
Ultrastructure of syndecans
 LAMININS

 Most important protein of lamina lucida.

 16 different laminins have been identified out of
which at least 4 are present in the skin in
significant quantities.
 They are heterotrimers. Made up of α, β & γ
subunits.
 There are 5 α subunits (α1-5), 3 β subunits (β1-3)
& 3 γ subunits (γ1-3).
 LM-111 denotes laminin - α1β1γ1.
 Consists of 3 domains :-
a) LN domain - Binds to other laminin molecules &
collagen.
b) Coiled-coil domain - Attaches the α, β & γ subunits.
c) LG domain – Binds with amino terminal end of
integrins.
 HOW THE LAMININ MOLECULES BIND WITH EACH
OTHER IN LAMINA LUCIDA ??
3 ARM INTERACTION HYPOTHESIS

Laminin molecules bind with each other by forming hexagonal

tesselations through their LN domain.
Laminin 332 – how it differs from other laminin molecules
APPLIED ASPECTS :
 Genetic mutations in any of the 3 polypeptide
units of laminin 332 can result in herlitz or non
herlitz junctional epidermolysis bullosa
with profound fragility of skin.
 Autoantibodies against laminin 332 causes
cicatricial pemohigoid.
 COLLAGEN
 Major collagen of lamina densa is collagen IV.
 Major collagen of reticular lamina is collagen III.

 All collagen molecule have a triple helical

structure & the basic structural unit is trimer of
polypeptide called tropocollagen.
 Multiple tropocollagen molecules aggregate to
form collagen fibrils which in turn cross linnk
extensively to form collagen fibres
 Individual polypeptide chain- left handed helix.

 Collagen monomer- right handed superhelix.

 Each turn of polypeptide chain consists of Gly-X-
Y, where most of the times X & Y are proline &
hydroxyproline respectively. Other important
aminoacids are lysine & hydroxylysine.
COLLAGEN BIOSYNTHESIS:
 Synthesized by dermal fibroblast, epidermal
keratinocytes, vascular endothelial cells &
smooth muscle cells.
COLLAGEN IV ORGANISATION IN LAMINA DENSA
APPLIED ASPECTS:
1) Hydroxylation of proline & lysine requires
ascorbic acid, molecular oxygen & ferrous iron
as co factors. So,
• In scurvy Deficiency of ascorbic acid

• Connective tissue weakness

• In chronic ulcers Poor circulation

Impaired collagen Low oxygen tension

production

Poor healing
2) Defect in α3, α4, α5, α6 chain of type IV collagen
causes different form of Alport’s syndrome.
3) Defect in α3 chain of type IV collagen causes
Goodpasture’s syndrome.
4) Mutation in type III collagen gene causes Ehler
Danlos syndrome.
 NIDOGEN
 Family of highly conserved sulfated monomeric
glycoproteins located in the basal lamina.
 Two types : Nidogen 1 & 2

 Structurally it (along with perlecan) connects the

networks formed by collagens and laminins to
each other.
 PERLECANS

 Basement membrane specific heparan sulfate

proteoglycan core protein (HSPG) or HSPG 2.
 Large multidomain proteoglycan that binds to &
crosslinks many ECM components & cell surface
molecules.
 It consists of a core protein & 3 long chains of
glycosaminoglycans ( often heparan sulfate but
can be chondroitin sulfate).
 Have 5 domains :

1) N- terminal domain I : site for attachment of

heparan sulfate chains.
2) Domain II : 4 repeats homologous to ligand binding
portion of the LDL receptor.
3) Domain III : Homologous to domain IV a & IV b of
laminin.
4) Domain IV : contains series of Ig modules.
5) C- terminal Domain V : homologous to LG domain
of laminin. Responsible for self assembly.
 FIBULINS

 Family of 6 highly conserved, calcium binding

ECM proteins.
 Located in blood vessels, basement membrane &
microfibrillar structures.
 Have binding sites for variety of ligands of both
basement membrane proteins & components of
interstitial connective tissues.
 Intermolecular bridges stabilizing elastic fibres &
basement membrane.
 Genetic defects of genes encoding fibuin 4 & 5
causes different forms of cutis laxa.
 BM 40 / SPARC
 Secreted protein acidic & rich in cysteine (SPARC)
or Basement membrane protein 40 ( BM 40) or
osteonectin.
 Consists of a single polypeptide chain having 4
domains :
1) Domain I ( amino terminal) : calcium binding
domain.
2) Domain II : cysteine rich domain.

3) Domain III : hydrophilllic region.

4) Domain IV ( carboxy terminus) : EF hand motif.

 Domain I & II are antigenic determinants.
 Domain III & IV have binding sites for collagen
IV.
 SPARC is an anti adhesive molecule & an
inhibitor of cell spreading.
 Plays a role in tumor cell invasion.

Increased sparc Decreased collagen

proliferation

Increaed tumor cell

invasion
Interaction of molecules in basal lamina
 ANCHORING FIBRILS
 Ultrastructurally recognisable, U – shaped
structures that extend from the lower part of the
lamina densa to the upper reticular dermis.
 Major components of anchoring fibrils are
collagen VII molecules.
 Individual collagen molecules consists of a
central triple helical collagenous domain flanked
by non helical globular domains i.e. NC1 at
amino terminal & NC2 at carboxy terminal.
o The large amino terminal non collagenous NC1 domain
interact with type IV collagen & laminin 332.

o Forms antiparallel dimers linked through their carboxy

terminal ends.

Organisation of collagen monomer to anchoring fibrils.

 Forms U shaped loops that entrap larger fibres in
a manner that stabilises the association of lower
part of the lamina densa to the upper papillary
dermis.
APPLIED ASPECTS :
 Complete absence of type VII collagen – Severe,
generalised, recessive dystrophic
epidermolysis bullosa.
 Missense mutations – Dominantly inherited
dystrophic epidermolysis bullosa.
 Transversion mutation – transient bullous
dermolysis of newborn.
 Autoantibodies against collagen VII –
Epidermolysis bullosa acquisita, Bullous
SLE.