Development of The Limb and

Muscle
Week II
FMS II
Dr Marselina I. Tan
 Learning objectives
Students should be able to:
• explain the embryological development of limb
• explain the embryological development of the
muscle
• know the abnormality
 Classification of mesoderm :
• Notochord: axial mesoderm

in adult substituted by columna vertebrate

• Somite : paraxial mesoderm

– Dermatome  Dermis
– Myotome  skeletal muscle
– Schlerotome  vertebrae, ribs

• Mesoderm intermediate : Urogenital system

• Mesoderm lateral :
– mesoderm splancnic : circulatory system
– mesoderm somatic : limbs
Derivatives of
mesoderm:
* Paraxial/
somite:
- Myotome
 skeletal
muscle
- Dermatome
- Schlereto
me 
vertebrae
* Intermediate
* Lateral
* Head
 • Expression patterns of
genes that regulates
somite differentiation.
Sonic hedgehog (Shh),
secreted by notochord
& floor plate of neural
tube  ventral part of
somite form sclerotome
& expressed PAX1
• PAX1 control
chondrogenesis &
vertebral formation.
• Wnt proteinactivate
PAX3 demarcates
dermomyotome.

Wnt prot  dorsomedial portion of somite form epaxial muscle &

express muscle specific gene Myf5.
Mid-dorsal portion of somite dermis by neurotropin 3 (NT-3)
expressed by dorsal neural tube.
Hypaxial musculature is derived from the dorsolateral portion of somite
under activating Wnt prot & BMP-4 prot  together activate MyoD
expression
Development of the Limb
 How do these abnormalities
arise?
THROUGH IMPROPER
DEVELOPMENT!!!

The goal of limb development research is to

understand how cartilage, muscle and tendons
are formed in the right place and how they make
the right connections with each other to form a
normal functional limb
 Why Study Limb
Development?
• Fundamental Understanding of the
process
• Clinical relevance
– Some statistics:
o Frequency of abnormality depends on severity
o Duplication of digits is fairly common at ~1 in
300 births
o Major failures of formation is less common, at
~ 1 in 100,000 births
(Indiana Hand Center)
 Limb Development is A Part
of Organogenesis
• The process of organogenesis involves
using the basic body plan (organized
embryo) to develop specific organs (limbs,
heart, eyes) in specific regions that leads
to the fully functional organism, capable of
independent survival
– Involves: pattern formation, specification of
positional information, induction,
morphogenesis, and cellular differentiation
 How are these processes
involved?
• Pattern formation  directs cell identity and
leads to ordered spatial pattern of cell activity
• Positional information  directs where organs
will form
• Induction  direction of specific cell fate
• Morphogenesis  changing the form of cells
• Differentiation  acquire functional and
structural identity distinct from their
surrounding cells
LIMB BUD FORMATION
 Limb Bud Formation

• The distal tip of the limb bud is covered with a transient

structure known as the apical ectodermal ridge (AER).
Limb outgrowth is dependent upon interactions between
the AER and the underlying mesoderm.

• If the AER is removed, the mesoderm stops dividing 

Therefore, limb outgrowth involves reciprocal interactions
between the AER and underlying mesoderm.

3/24/2009
What Controls Establishment of Limb Axes?
• The limb has three axes:
– Proximo-distal;
– Anterior-posterior and
– Dorsal-ventral.

• Proximo-distal
Removal of the AER at progressively later stages results in a
truncated limb with progressively more distal elements. Thus, these
elements are laid down in a proximal-to-distal direction.
The AER does not regulate P-D polarity. Fates of cells are
determined by the length of time they spend in the progress zone.
Those residing there the longest become the most distal elements.
 Development of the Limb
• As the limb bud grows outward,
cells in the progress zone proliferate
and acquire a positional value
•When cells leave the progress zone,
cartilage may begin to differentiate,
and other elements gain their
positional information
 Anterior-Posterior Axis
This axis is determined by the zone of polarizing activity (ZPA),
which is located at the junction between the limb bud and the
body wall.
Grafts of the ZPA to the anterior margin of a host limb bud causes
duplication of digits in mirror-image symmetry.
A diffusible morphogen (retinoic acid?) is released from the ZPA.

3/24/2009
 Retinoic Acid : A Developmental
Morphogen

• Retinoic acid is the active form of Vitamin A

• Both excess retinoic acid and vitamin A

deficiency each affect left-right symmetry
(e.g., abnormal formation of the mammalian
heart) through alterations in gene
expression.
Pengatur arah perkembangan anggota
tubuh
Gen-gen yang berperan dalam pembentukan anggota tubuh
• Sonic Hedgehog (Shh)
• Fibroblast Growth Factor 4 (FGF-4)
– Gen Shh mengkode molekul sinyal
– Anggota dari keluarga FGF
induksi yang berperan selama
embriogenesis – Menstimulasi sel-sel mesoderm
dan mengarahkan sinyal untuk
– Berperan menentukan nasib sel perkembangannya
– Sinyal utama penentu sumbu – FGF-4 -/-  embrio mati
antero-posterior – Pada jaringan dewasa tidak
– Alx-4 menekan ekspresi Shh aktif ditranskripsikan
Cell Death and Digit Formation

Without apoptosis, the fingers would be webbed: syndactyly

• Kematian sel
apoptosis pada
pembentukan jari
DEVELOPMENT OF MUSCLE:
THE MYOTOMES
Figure 10.1 Cross-sectional drawings showing the stages of development in a somite. A. Mesoderm cells become
epithelial and are arranged around a small lumen. B. Cells in the ventral and medial walls of the somite lose their
epithelial characteristics and migrate around the neural tube and notochord, and some move into the parietal layer
of lateral plate mesoderm. Collectively, these cells constitute the sclerotome. Cells at the dorsomedial (DML) and
ventrolateral (VLL) regions of the somite form muscle cell precursors. Cells from both regions migrate ventral to
the dermatome to form the dermomyotome. VLL cells also migrate into the parietal layer of lateral plate mesoderm
across the lateral somitic frontier (green line). In combination, somitic cells and lateral plate mesoderm cells
constitute the abaxial mesodermal domain, while the primaxial mesodermal domain only contains somitic cells
(paraxial mesoderm). C. Together, dermatome cells and the muscle cells that associate with them form the
dermomyotome. D. The dermomyotome begins to differentiate: Myotome cells contribute to primaxial muscles,
and dermatome cells form the dermis of the back.
REGION SPECIFIC OF
MYOBLAST
 Limb malformation

These are clenched hands resulting from

camptodactyly (fingers bent over) and
clinodactyly (fingers inclined to one side or the
other). This particular appearance is very
suggestive of trisomy 18.
Deformation from constraint with
oligohydramnios in utero can result in the
appearance of the club feet (varus deformity)
seen here. The feet are turned inward.
Example of abnormality  absent of
hands (+ legs)
(amelia/ phocomelia)  caused by
thalidomide (anti-vomiting)

Syndactyly represents fusion of two or more

digits. It can be an isolated finding or part of
syndromes that define patterns of anomalies.
Most of these syndromes do not have a specific
genetic defect yet defined, though some do.
Seen above are the digits of the hand fused
together. Below, the foot has rudimentary,
partially fused digits.
 This is sirenomelia. The term comes from "siren" or
"mermaid" because of the characteristic fusion of the lower
extremities that results from a failure in the development of
a normal vascular supply to lower extremities from the lower
aorta in utero.

Regulative
ability of Limb
Sirenomelia (caudal dysgenesis) field
gastrulation disruption. Loss of
mesoderm in the lumbosacral
region has resulted in fusion of limb
buds and other defects.
 What are these
abnormalities?

Polydactyly – the
duplication of a digit
 Caution: aspirin; streptomycin
(deafness); antidepressant (limb
deformities); tetracyclin (bone and
tooth anomalies) etc
Cocaine, LSD, PCP (CNS, limb
abnormalities, hypoxia)

Child with unilateral amelia

Figure 10.6 Poland sequence. The pectoralis
minor and part of the pectoralis major muscles
are missing on the patient's left side. Note
displacement of the nipple and areola.
Figure 10.7 Prune belly syndrome: a
distended abdomen from atrophy of
abdominal wall musculature.
Figure 10.8 Infant with arthrogryposis
(congenital joint contractures).