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 & 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 reticularis/ Reticular lamina : Basal lamina is
attached with reticular lamina through the anchoring
fibrils/ filaments.
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 PROTEINS
INHERITANCE
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.

 COOH
terminal

Laminin

NH2 terminal

Interaction of integrin with actin & laminin

 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.
 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.
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.
 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.
ULTRASTRUCTURE OF HEMIDESMOSOME
 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.
 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.
Ultrastructure of syndecans
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 :-
• 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.
 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.
 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 ??

Laminin molecules bind with each other by forming hexagonal tesselations

through their LN domain.
APPLIED ASPECTS :
 Genetic mutations in any of the 3 polypeptide units of
laminin 332 will result in junctional epidermolysis
bullosa with profound fragility of skin.
 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.
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 chain of type IV collagen causes Alport’s
syndrome.
3) Defect in α5 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.
 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.
 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.
 Autoantibodies against collagen VII – Epidermolysis
bullosa acquisita, Bullous SLE.