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INDIAN DENTAL ACADEMY

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DEVELOPMENTAL BIOLOGY
GROWTH
DIFFERENTIATION
MORPHOGENESIS
Developmental biology is the study of the process by
which organisms grow and develop.
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GROWTH
It is the physiochemical process by which an
organism becomes larger.
[Salzmann]

Growth usually refers to an increase in size or
number. It is largely an anatomic phenomenon.
[Proffit]


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A normal process of increase in size of an
organism as a result of accretion of tissue
similar to that originally present.


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DIFFERENTIATION
Cellular differentiation is a concept from
developmental biology describing the process
by which cells acquire a "type".

The morphology of a cell may change
dramatically during differentiation, but the
genetic material remains the same.
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A cell that is able to differentiate into many
cell types is known as pluripotent.

These cells are called stem cells in
animals.

A cell that is able to differentiate into all cell
types is known as totipotent. In mammals,
only the zygote and early embryonic cells
are totipotent.
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Embryonic stem
cells are stem cells
derived from the
inner cell mass of an
early stage embryo
known as a
blastocyst.

Human embryos reach the blastocyst stage
4-5 days post fertilization, at which time they
consist of 50-150 cells.
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Adult stem cells are undifferentiated cells
found throughout the body that divide to
replenish dying cells and regenerate
damaged tissues. Also known as somatic
stem cells.

Umbilical cord blood is human blood
from the placenta and umbilical cord that
is rich in hematopoietic stem cells.
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Derived from the Greek
word morph shape and
genesis creation


It is one of three fundamental aspects of
developmental biology along with the control
of cell growth and cellular differentiation.

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Morphogenesis is concerned with the
shapes of tissues, organs and entire
organisms and the positions of the
various specialized cell types.

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MOLECULAR BASIS OF
CRANIOFACIAL DEVELOPMENT
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The development of the
craniofacial region is an
amalgamation of complex
processes that need to be
coordinated to produce a
working unit such as a
calvarium, cranial base, or
palate.
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A common theme in the
early development of
craniofacial structures is
balancing the level of
progenitor populations by
growth factor signaling.

This is to form a large enough
population of progenitor cells
before differentiation, or that
the proliferation is maintained
for the outgrowth of the
element.
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What is the Molecular
basis of growth
?
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Several types of molecules are particularly
important during growth, differentiation and
morphogenesis.

Morphogens are soluble molecules that can
diffuse and carry signals that control cell
differentiation decisions in a concentration-
dependent fashion.
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MORPHOGENS typically act
through binding to specific
protein receptors. An
important class of molecules
involved in morphogenesis
are transcription factor
proteins that determine the
fate of cells by interacting with
DNA.

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These can be coded for by master regulatory
genes and either activate or deactivate the
transcription of other genes and, in turn,
these secondary gene products can regulate
the expression of still other genes in a
regulatory cascade.


Another class of molecules involved in
morphogenesis are molecules that control
cell adhesion.
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For all intents and purposes, craniofacial
development is initiated as soon as the
anteroposterior axis of an embryo is established

The ability to specify a head structure, rather than
reiterate another body segment, was a crucial step in
vertebrate evolution that corresponded to the
acquisition of two cell populations: the neural crest
and the ectodermal placodes

(reviewed by Basch et al., 2004 ; McCabe et al.,
2004 ).
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In recent years, new data have begun to
reveal how the neural crest cell population is
actually generated, what types of controls are
in place to modify neural crest cell migration
and, ultimately, the role that this cell
population plays in establishing the pattern of
the craniofacial skeleton.

Although the neural crest receives a
significant amount of attention, it is not the
only craniofacial tissue with patterning
information.
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This seminar today focuses on innovative
studies that have addressed these issues,
sometimes with new and unexpected
results.
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In the beginning
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Although the postnatal
vertebrate head exhibits
an exceedingly intricate
and varied morphology,
the craniofacial complex
initially has a much more
simple geometry.


It consisting of a series of swellings or that undergo
growth, fusion and expansion.

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There are seven prominences that comprise the
vertebrate face.
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Until recently, it was thought that the ventral
region of the first pharyngeal (branchial) arch
gave rise to the mandibular prominence and
therefore the lower jaw.

It was also believed that the dorsal region of
the first arch gave rise to the maxillary
prominences, which form the sides of the
middle and lower face, the lateral borders of
the lips, and the secondary palate.
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Two new studies, carried out in avians and
axolotls, contest this view and demonstrate
that at least part of this is incorrect.

Both groups show that the ventral region of
the first arch actually gives rise to both
maxillary and mandibular skeletal elements,
rather than to only the mandibular elements,
as previously thought.
(Cerny et al., 2004 ; Lee et al., 2004 )
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When considering the origami-like process of
tissue folding, flexure and growth that ultimately
results in a face, one must also bear in mind that
the cells comprising the face have undergone a
massive relocation.

This relocation occurs owing to both active neural
crest cell migration and the passive displacement
of tissue that is associated with neurulation and
head flexure.
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NEURULATION
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It is the process of development of the

NEURAL PLATE results from the
thickening of the ectoderm overlying the
Notochord.

NEUROECTODERM cells comprising the
neural plate.

Folding to produce NEURAL TUBE lateral
edges of the neural plate get elevated to form
the neural folds
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Neurulation begins with a unified layer of
ectoderm, underneath which lies the
endoderm.
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Ectomeres are discrete regions of superficial
ectoderm that exhibit a segmented pattern of
gene expression.

Together with neural crest and neuro
ectoderm, they define a larger developmental
unit .
(Couly and Le Douarin, 1999).

Later, these tissues act on signaling centers
in the facial prominences
(Hu et al., 2003 ).
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The ectoderm begins to fold upwards, giving rise to
the neural folds. During this process, interactions
between signaling molecules begin to delineate the
medial ectoderm as being neural (green) and the
lateral regions of ectoderm as being non-neural (blue).
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The neural tube forms upon fusion of the neural folds,
giving rise to discrete neuroectoderm (green) and
surface ectoderm (blue). Around the same time, the
border region between the neuroectoderm and
surface ectoderm gives rise to neural crest cells.
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Neurulation completes upon formation of the neural
tube, and neural crest cells (nc) lie sandwiched
between the facial (surface) ectoderm and the
neuroectoderm.
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Sagittal section through neural tube of a
chick embryo, showing neural crest (nc)
located between surface ectoderm (se) and
neuroectoderm (ne). L, lateral; M, medial.
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The neural tube is the primodium of the C.N.S . The
anterior region forms the Fore brain, Hind brain
AND Mid brain. Eight bulges form in the hind brain
known as RHOMBOMERES.

Neural crest cell which arise from the neural folds
migrate through out the body to form varied
structures. They migrate from each rhombomere to
a specified location.

Neural crest cells needed for development of face
and first pharyngeal arch structures arise from
MID BRAIN and RHOMBOMERES 1,2 & 3.
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NEURAL CREST MIGRATION
AND
ECTODERM ALIGNMENT
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As the closed neural tube begins to differentiate
into the central nervous system, the neural
crest begins to migrate anteriorly from specific
rhombomeres (r1-r3) into discrete regions of the
face.
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During this process, the neuroectoderm (ne)
and surface ectoderm (se) components of the
ectomeres continue to remain aligned (yellow
arrows in C).
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As neural crest migration nears completion, the
neuroectoderm and facial ectoderm are no
longer aligned.
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CREATING THE
NEUROECTODERM-SURFACE
ECTODERM BOUNDARY
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One of the first crucial steps in craniofacial
development occurs when head ectoderm
is subdivided into non-neural and neural
regions

This effectively establishes which head
epithelium will lie outside of the cranial
neural crest and which will lie inside it.
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A subset of epithelial cells located at this
neural/non-neural boundary separate from
the epithelium, adopt a mesenchymal
character and come between these two
epithelia as they start their migration.

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The epithelial-mesenchymal transition that
marks the birth date of the neural crest, this
shift is upon bone morphogenetic protein (Bmp)
signaling.

When Bmp signaling is inhibited by the
overexpression of noggin, a Bmp antagonist,
this transition is blocked and neural crest cells
are no longer generated from the margins of the
neural folds
(Burstyn-Cohen et al., 2004 ).
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Noggin is a polypeptide that binds to members
of the TGF- superfamily of proteins. It is a Bone
morphogenetic protein inhibitor.
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Bmp signaling achieves this effect in part by
regulating Wnt1 gene


In turn, Wnt signaling appears to be essential
for the generation of neural crest cells as
inhibition of its activity can block the
production of neural crest cells (Garcia-Castro
et al 2005)

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In addition to Bmp and Wnt proteins, several
new molecules have also been implicated in
the generation or early migration of neural
crest cells.


Sox transcription factors, which are well
known for their roles in skeletogenic cell fate
and sex determination, are also involved in
generating neural crest cells (Cheung and
Briscoe, 2003 ; Perez-Alcala et al., 2004 ).
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These studies indicate that the overexpression
of Sox genes lengthens the developmental
window during which cranial and trunk neural
crest cells can be induced, and then promotes
neural crest-like characteristics in those cells.

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Neural crest contributions to
craniofacial patterning
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Which tissue controls facial patterning?

The answer to this question continues to be
debated. In two recent studies, the contribution
of the neural crest to facial patterning was
assessed by swapping neural crest cells
between ducks and quails.
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It was found that switching frontonasal
neural crest cells between ducks and
quails resulted in alterations to such an
extent that ducks with quail frontonasal
neural crest cells had a quail-like beak,
and quails carrying duck neural crest cells
had a duck-like beak.
(Schneider and Helms, 2003 ).

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Tucker and Lumsden reached a near-
identical conclusion when they independently
performed the same types of inter-species
transplants.

They, too, found that the capacity to form
species-specific skeletal elements in the head
is an inherent property of the neural crest
cells.
(Tucker and Lumsden, 2004 )
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It should be emphasized that in both these
studies, the extent to which facial features
were transformed was directly proportional
to the number of transplanted neural crest
cells.

In other words, the transformation was a
`population-dependent' effect, as was
reported in much earlier transplantation
studies (Andres, 1949 ).

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So it seems that only when the contingency
is large enough do neural crest cells follow
molecular cues that are generated and
maintained by the assemblage itself,
disregarding signals emanating from the
local environment.

When the numbers of transplanted cells are
below some crucial threshold, then they
appear to respond to local cues from the
surrounding epithelia.

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Just what these population-dependent cues
are, and how many cells are required to
maintain them, is unknown.

What we do know, however, is that facial
morphogenesis is the cumulative result of
reciprocal signaling between and among all of
these tissues, and that the debatable issue of
which tissue contains patterning
information becomes a question of timing.

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Preview of next seminar:
Epithelial contribution to craniofacial patterning
Oral ectoderm and tooth patterning
Pharyngeal endoderm and arch patterning
Neural and surface ectoderm: patterning the middle
and upper face
Molecular mediators of craniofacial morphogenesis
A sonic boom
HOX genes
Fgfs and craniofacial patterning: a question of timing
Bmp proteins and craniofacial patterning
Bmp4 and craniofacial patterning
Patterning of the jaw


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THANK YOU . . .
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Presented by :
Dr. Mahima Nanda
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No region of our anatomy more powerfully
conveys our emotions nor elicits more
profound reactions when disease or
genetic disorders disfigure it than the face.
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Recent progress has been
made towards defining the
tissue interactions and
molecular mechanisms that
control craniofacial
morphogenesis.


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Some insights have come
from genetic manipulations
and others from tissue
recombination and
biochemical approaches,
which have revealed the
molecular underpinnings of
morphogenesis.

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Changes in craniofacial architecture also
lie at the heart of evolutionary adaptation,
as new studies in attest.

Together, these findings reveal much
about molecular and tissue interactions
behind craniofacial development.
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Sometimes, the mechanisms that regulate
normal development are best appreciated
by studying cases of abnormal
development.


Human craniofacial malformations have
been avidly catalogued since the
Aristotelian era but only lately have
researchers pinpointed some of the genes
responsible.

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The next hurdle is to understand the function of
the encoded proteins in craniofacial
morphogenesis.

This aim is complicated by the fact that these
genes are invariably expressed in multiple
tissues and at multiple times during facial
development, and so separating their numerous
functions becomes a difficult task.

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The importance of genes on development
has been know for a long time.

In Drosophila a class of Master regulatory
genes were know to exist long before
molecular basis was known.

Genes known to control segment
differentiation were called Homeotic
Genes.
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Mutations of these genes caused parts to
develop in inappropriate places.

This process of transformation was called
HOMEOSIS by Bateson back in the
1940s.

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Here, a mutation of the Antennapedia gene
causes a leg to develop in place of the
antenna. Note the little Haltere (balancer) on
the segment behind the wing.
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The next slide shows homeotic mutation
that results in transformation of this haltere
into a second wing.
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Molecular mediators of
craniofacial morphogenesis
Development 132, 851-861 (2005)
Helms JA, Cordero D, Tapadia MD.
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A sonic boom
HOX genes
Fgfs and craniofacial patterning: a
question of timing
Bmp proteins and craniofacial
patterning
Bmp4 and craniofacial patterning

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A SONIC BOOM !!!!
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The hedgehog gene (hh) was first identified
in the classic Heidelberg screens of Eric
Wiechaus and Christiane Nusslein-Volhard,
as published in 1978.

These screens identified genes that control
the segmentation pattern of Drosophila
melanogaster (fruit fly) embryos.
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The hh loss of function
mutant phenotype
causes the embryos to
be covered with
denticles (small pointy
projections), much like
a hedgehog.

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Investigations aimed at finding a hedgehog
equivalent in mammals revealed three
homologous genes.

The first two discovered, desert hedgehog and
Indian hedgehog, were named for species of
hedgehogs.
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Sonic hedgehog
was named after
Sega's video game
character Sonic the
Hedgehog.

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PROCESSING
SHH undergoes a series of processing
steps before it is secreted from the cell.
Newly synthesised SHH is referred to as
the PREPROPROTEIN. www.indiandentalacademy.com
As a secreted protein it contains a short
signal sequence at its N-terminus, which is
recognised by the signal recognition particle
during the translocation into the
endoplasmic reticulum (ER).

Once translocation is complete, the signal
sequence is removed by signal peptidase in
the ER.
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There SHH undergoes autoprocessing to
generate a N-terminal signaling domain
(SHH-N) and a C-terminal domain with no
known signaling role.

The cleavage is catalysed by a protease
within the C-terminal domain. During the
reaction, a cholesterol molecule is added
to the C-terminus of SHH-N.
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Thus the C-terminal domain acts as an intein
and a cholesterol transferase.

Another hydrophobic moiety, a palmitate, is
added to the alpha-amine of N-terminal
cysteine of SHH-N.

This modification is required for efficient
signaling, resulting in 30-fold increase in
potency over the non-palmitylated form

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HEDGEHOG SIGNALING PATHWAY.
HEDGEHOG SIGNALING PATHWAY.
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When SHH reaches its target cell, it binds to
the Patched-1 (PTCH1) receptor.

In the absence of ligand, PTCH1 inhibits
Smoothened (SMO), a downstream protein
in the pathway.

The binding of SHH relieves SMO inhibition,
leading to activation of the GLI transcription
factors: the activators Gli1 and Gli2 and the
repressor[Gli3].
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Activated GLI accumulates in the nucleus
and controls the transcription of hedgehog
target genes.

Overexpression of mutated PTCH1 plays
a role in basal cell carcinoma.
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FUNCTION
Of the hh homologues, shh has been
found to have the most critical roles in
development, acting as a morphogen
involved in patterning many systems,
including the limb and midline structures in
the brain and spinal cord.
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Members of the hedgehog family play key
roles in a wide variety of developmental
processes.

One of the best studied examples is the
action of Sonic hedgehog during
development of the vertebrate limb.
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One of the best studied craniofacial abnormalities
is holoprosencephaly (HPE), a syndrome that is
associated with perturbations in a handful of Shh-
related genes.

At one end of the HPE spectrum, fetuses exhibit
cyclopia, a condition characterized by a single,
central eye and no discernable nose, but a
relatively normal-looking middle and lower face
(Chiang et al., 2001 ).

At the other extreme, obligate HPE carriers can
have a normal facial appearance (McKusick, 2000
).
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In an effort to explain this remarkable phenotypic
variation, Traiffort and colleagues recently
examined how specific human HPE mutations
affected the structure and function of the SHH
protein.

The researchers found that most HPE mutations
fall into one of three classes:
1. mutations that influenced binding of the protein
2. those that affect the auto-processing of SHH
3. and those that adversely alter SHH stability
(Traiffort et al., 2004 ).
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Holoprosencephaly is a type of cephalic disorder.

This is a disorder characterized by the failure of the
prosencephalon (the forebrain of the embryo) to
develop.
During normal development the forebrain is formed
and the face begins to develop in the fifth and sixth
weeks of human pregnancy.
Holoprosencephaly is caused by a failure of the
embryo's forebrain to divide to form bilateral cerebral
hemispheres.

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Alobar holoprosencephaly - the most
serious form in which the brain fails to
separate, is usually associated with severe
facial anomalies.

Semilobar holoprosencephaly - in which
the brain's hemispheres have a slight
tendency to separate, is an intermediate form
of the disease.

Lobar holoprosencephaly - in which there
is considerable evidence of separate brain
hemispheres, is the least severe form.

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Alobar holoprosencephaly
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However, none of these mutation types could
be linked to a specific phenotype
(Traiffort et al., 2004 )

If there is no clear genotype-phenotype
correlation, then what explains the variable
expressivity of this craniofacial malformation?
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One appealing hypothesis is that environmental
agents act in conjunction with an autosomal
dominant mutation to compromise Shh
signaling.
(Cordero et al., 2004 ; Edison 2003 ).

If this scenario were true, then varying the time
in which an embryo was exposed to an
environmental teratogen could elicit different
disease phenotypes.

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Testing this hypothesis
Chen et al., 2002
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They exposed avian embryos to cyclopamine,
a potent inhibitor of the Hedgehog signaling
pathway ..

and found that by varying the delivery time so
that it coincided with Shh induction in the
forebrain and later in the face, we could
reproduce the spectrum of HPE phenotypes.


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Although this is unlikely to be the sole, or
even the predominant, explanation for
variations in HPE phenotype, experiments
such as these indicate that Shh has a
variety of functions in facial development.
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Presented by :
Dr. Mahima Nanda
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Molecular mediators of
craniofacial morphogenesis
continued . . . www.indiandentalacademy.com
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The central dogma of
molecular biology
describes the two-step
process, transcription
and translation, by
which the information
in genes flows into
proteins:

DNA RNA protein
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The basic building block of a protein is
the amino acid.
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HEDGEHOG SIGNALING PATHWAY.
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Fgfs and craniofacial patterning
: a question of timing
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Fibroblast growth factors, or FGFs, are a
family of growth factors involved in wound
healing and embryonic development.

The FGFs are heparin-binding proteins and
interactions with cell-surface associated
heparan sulfate proteoglycans have been
shown to be essential for FGF signal
transduction
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Fibroblast growth factor was found in a
cow brain extract by Gospodarowicz and
colleagues and tested in a bioassay which
caused fibroblasts to proliferate (first
published report in 1974).

In humans, 20 members of the FGF family
have been identified all of which are
structurally related signaling molecules:
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1. Members FGF1-10 all bind fibroblast growth
factor receptors (FGFRs). FGF1 is also
known as "Acidic", and FGF2 is also known
as basic FGF.

2. Members FGF11-14 are involved in
intracellular processes unrelated to the
FGFs

3. Members FGF16 through FGF23 are newer
and not as well characterized.

4. FGF15 is the mouse ortholog of human
FGF19
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One of the most important functions of
bFGF (FGF2) is the promotion of
endothelial cell proliferation and the
physical organization of endothelial cells
into tube-like structures.

It thus promotes angiogenesis, the growth
of new blood vessels from the pre-existing
vasculature.
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bFGF is an important player in wound
healing. It stimulates the proliferation of
fibroblasts that give rise to granulation
tissue, which fills up a wound space/cavity
early in the wound healing process.
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It has also been demonstrated that fibroblast
growth factors are associated with many
developmental processes in the craniofacial
region.

This has been well illustrated in recent
studies evaluating the consequences of Fgf
perturbation at four separate points in
craniofacial development.
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Early in craniofacial development, Fgf signaling
is crucial in establishing the midbrain-hindbrain
boundary
(Scholpp et al., 2003)

Later in development, Fgf signaling from ventral
forebrain and pharyngeal endoderm is required
for pharyngeal skeletogenesis, as inhibiting this
pathway prevents the formation of the second
arch skeleton
(Creuzet et al., 2004 ; Mason, 2003 )
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Later still, blocking Fgf signaling from the
surface ectoderm disrupts outgrowth of the
frontonasal skeleton
(A. Abzhanov, D. Hu, J. Sen, C. J)


Finally, just before birth, disruptions in Fgf
signaling cause premature osteogenesis in
the sutures.
(Moore et al.; Sarkar et al. )
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FGF-5 in the embryo is notable for its highly
specific pattern of expression, first in pre-
gastrulation embryonic ectoderm and later in a
small patch of mesoderm through which the
hepatic bud will penetrate.

Ectodermally-derived FGF-8 is suggested to
be important in driving the proliferation of the
underlying mesenchyme, thus adding length to
the limb.
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Clearly then, Fgfs play multiple, fundamental
roles in craniofacial morphogenesis, but
unraveling this complicated molecular
machinery will have to await better genetic
and molecular tools that permit a more precise
regulation of gene activity.
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What is a homeobox?
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A homeobox is a DNA sequence found
within genes that are involved in the
regulation of development (morphogenesis)
of animals, fungi and plants.

Genes that have a homeobox are called
homeobox genes and form the homeobox
gene family.
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Discovery

They were discovered independently in
1983 by Walter Jakob Gehring and his
colleagues at the University of Basel,
Switzerland.
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Since their discovery in 1983, homeobox
genes, and the proteins they encode, the
homeodomain proteins, have turned out to
play important roles in the developmental
processes of many multicellular organisms.


While certainly not the only developmental
control genes, they have been shown to play
crucial roles from the earliest steps in
embryogenesis to the very latest steps in cell
differentiation.
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HOX GENES
Hox genes are a subgroup of homeobox
genes.

In vertebrates these genes are found in
gene clusters on the chromosomes.

In mammals four such clusters exist,
called Hox clusters.
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The gene name "Hox" has been restricted
to name Hox cluster genes in vertebrates.

Only genes in the HOX cluster should be
named Hox genes.

So note: homeobox genes are NOT Hox
genes, Hox genes are a subset of
homeobox genes.

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HOX cluster
The term Hox cluster refers to a group of
clustered homeobox genes, named Hox
genes in vertebrates, that play important
roles in pattern formation along the anterior-
posterior body axis.

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Homeodomain: a DNA-binding domain,
usually about 60 amino acids in length,
encoded by the homeobox.


Homeobox: a fragment of DNA of about
180 basepairs (not counting introns),
found in homeobox genes.

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Here a rotating view of the homeodomain
bound to DNA
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HUMAN GENES
NAME CHROMOSOME GENE
HOX A Chromosome 7
HOXA 1-7
HOXA 9-11,
HOXA 13
HOX B Chromosome 17
HOXB 1-9
HOX C Chromosome 12
HOXC 4-6
HOXC 8-13
HOX D
Chromosome 2

HOXD 1
HOXD 3-4
HOXC 8-13
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If we look at the CNS earlier in embryogenesis, shortly
after neurulation, and during somitogenesis,
segmentation is obvious in the rhombencephalon, which
is the CNS precursor of the hindbrain.

In this structure and at this time there is a segmental
periodicity to the expression of the most anteriorly
expressed genes.

So, each rhombomere has its own hox code as well
(i.e., except R1 and R2, which do not express hox
genes. .
Hox Gene Expression and
Embryogenesis
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Key Features of this are :
1) The vertebrate homeotic complex comprises four
distinct Hox gene clusters (Hox A, B, C, D).

2) The chromosomal organization of the genes in each
Hox cluster reflects its anterior-posterior expression in
the body plan (spatial colinearity).

3) Homeotic genes are expressed within segmented
and unsegmented structures within the body plan. Hox
gene expression in some unsegmented structures
arise from segmented precursors.
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Bmp proteins and
craniofacial patterning
Initial discovery of bone morphogenetic
protein activity was published in 1965
by Marshall R
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Bone Morphogenetic Proteins (BMPs) are a
group of growth factors known for their ability
to induce the formation of bone and cartilage.

TYPES

Originally, seven such proteins were
discovered, 6 of them (BMP2 through BMP7)
belong to the Transforming growth factor beta
superfamily of proteins.
Since then, nine more BMPs have been
discovered, bringing the total to sixteen.
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FUNCTION
BMPs interact with specific receptors on the cell
surface, referred to as bone morphogenetic protein
receptors (BMPRs).

Signal transduction through BMPRs results in
mobilization of members of the SMAD family of
proteins.

The signaling pathways involving BMPs, BMPRs
and SMADS are important in the development of
the heart, central nervous system, and cartilage, as
well as post-natal bone development.
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They have an important role during embryonic
development on the embryonic patterning and
early skeletal formation.

As such, disruption of BMP signaling can affect
the body plan of the developing embryo. For
example, BMP4 and its inhibitors noggin and
chordin help regulate polarity of the embryo (i.e.
back to front patterning).
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Mutations in BMPs and their inhibitors
(such as sclerostin) are associated with a
number of human disorders which affect
the skeleton.

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Tabin and co-workers set out to understand
how such morphological variations might
arise due to BMPs.

(2004)

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They evaluated two finch species - the ground
and cactus finches - that represent the
extremes in Galapagos finch beak morphology
(Grant, 1986 ).
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At the time when ground and cactus finch
embryos appear similar, in situ hybridization
analyses by these investigators revealed a
difference in the patterns of Bmp4 expression.
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Epithelial contribution to
craniofacial patterning
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Neural and
surface
ectoderm:
patterning
the middle
and upper
face
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When regions of facial ectoderm are
transplanted to ectopic sites in the avian face,
the developmental fate of underlying frontonasal
neural crest cells is altered and the result is a
duplication of upper beak structures (Hu et al.,
2003 ).

This same bit of facial ectoderm can elicit similar
duplications when transplanted into the first,
Hox-negative, arch, but has no effect when
transplanted into the second, Hox-positive, arch
(Hu et al., 2003 ).
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This result indirectly illustrates how neural
crest plasticity is balanced against a `pre-
pattern', owing in no small part to the
expression of Hox genes in the facial
tissues (Creuzet et al., 2002 ).

What types of signals imbue this facial
ectoderm with the ability to re-specify the
fates of neural crest cells?

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Both Shh and Fgf8 are expressed in this
region of tissue, but whether they are the
molecules responsible for achieving this
effect, or simply molecular markers of an
important boundary domain in the face,
remains to be determined.

Neural ectoderm is also a source of
patterning information for the middle and
upper face, as has recently been shown in
a series of experiments conducted in
zebrafish.
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In these experiments, it was found that
Shh emanating from anterior ventral
neuroectoderm directly patterned the
ventral surface ectoderm, without requiring
an intermediate signal generated by neural
crest.
The loss of neuroectodermal Shh
prevented neural crest cells from
aggregating into condensations and
eventually from forming skeletal elements.

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Preview of next seminar:
Molecular mediators of craniofacial morphogenesis
HOX genes

Epithelial contribution to craniofacial patterning
Oral ectoderm and tooth patterning
Pharyngeal endoderm and arch patterning
Neural and surface ectoderm: patterning the middle and
upper face

Patterning of the jaws
Maxilla
Mandible
palate


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THANK YOU . . .
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At the time when ground and cactus finch embryos appear similar,
in situ hybridization analyses by these investigators revealed a
difference in the patterns of Bmp4 expression (Abzhanov et al.,
2004 ) (see Fig. 4). To test experimentally whether spatial and
temporal changes in Bmp4 expression could account for the relative
size and shape differences in these finches' beaks, the investigators
mis-expressed Bmp4 throughout the mesenchyme of a chick
frontonasal prominence (Fig. 4D). This misexpression converted the
narrow short chick beak into a much broader bigger beak that
resembled that of the large ground finch (Abzhanov et al., 2004 )
(Fig. 4D).

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Vertebrates exhibit a marvelous range of craniofacial
features that are designed to fit specialized niches and
behaviors.
These postnatal facial features are immediately obvious,
but during the embryonic period, vertebrate faces look
remarkably similar (Haeckel, 1897 ).
The proteins that establish this basic blueprint of the
craniofacial region are still unidentified but likely
candidates are those same molecules that establish
other developmental axes in vertebrates and
invertebrates: Hedgehog and Wnt proteins, and
members of the Bmp and Fgf families. Some new
studies have begun to explore how different species use
these pathways to create distinctive facial features.
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In the Galapagos finches, Darwin had noted that `a
nearly perfect gradation may be traced from a beak
extraordinarily thick to one so fine that it may be
compared with that of a warbler.' (Darwin, 1859 ). We
now know that these species-specific morphological
variations are evident during embryogenesis, and are
first evident around Hamburger and Hamilton
(Hamburger and Hamilton, 1951 ) stage 22 (S.
Brugmann and J.A.H., unpublished). Prior to that time,
the faces of different avian species are indistinguishable
from one another (Schneider and Helms, 2003 ). Tabin
and co-workers set out to understand how such
morphological variations might arise.
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Epithelial contribution to
craniofacial patterning
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PRE-NATAL CRANIOFACIAL
MORPHOGENESIS
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Basic research concerning craniofacial
development runs along 2 pathways
namely the molecular and the
morphometric. This gap will now be
bridged in the initial part if this seminar
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Pre natal craniofacial
morphogenesis : 4-D visualization
of morphogenetic processes
Orthod Craniofacial Res 6( sppl. 1), 2003;89-94
Radlanski RJ
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Using histological sections of human fetuses
computer aided 3-D reconstuctions were
made with special focus given to all
anatomical structures of orofacial region of
the orofacial region of the growing head.
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The hard tissues like the
bones, cartilages, and teeth,
play many essential roles for
our survival.

Calvarial and cranial base
bones protect the brain and
the sense organs from
external shocks. Three small
bones in the inner ear are
needed for us to hear. Jaws
allow us to talk and along with
the teeth allow us to chew.

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