EARLY EMBRYOLOGY: SOMITE STAGE AND LIMB BUDS

Week 1-2: formation of zygote, implantation and formation of bilaminar embryo.

Weeks 3-8: Embryological period.
Weeks 9-38: Fetal period.

DEVELOPMENT OF THE SOMITES (week 3)

The intraembryonic mesoderm on each side of the forming notochord and neural tube thickens to form
a longitudinal column of paraxial mesoderm. By the end of the 3rd week, the paraxial mesoderm divides
into paired bodies called somites, located bilaterally of the neural tube.
Somites

The somites give rise to the axial skeleton (vertebrae, ribs), associated musculature and adjacent
dermis of skin.
The first pair of somites develop a short distance posterior to the cranial end of the notochord,
and the rest of the somites form caudally. Around 38 pairs of somites form during the somite
period of development, from days 20 to 30. The final number is 42 to 44 pairs. The somites may
be used as a criterion to determine the age of the embryo.
A cavity, the mycocoele, forms within each somite but disappears.
Each somite becomes differentiated into ventromedial sclerotome (for vertebrae and ribs),
myotome (muscles) and dermatome (skin).

Week 4

At the beginning of the 4th week, the somites (4) are well formed and the neural tube is also
formed but it is opened at the rostral and caudal neuropores.
Upper limb buds become recognizable during week 4 (day 26 or 27) and the lower limb buds
become present by the end of week 4 (day 28). The patterning of the limb development is
regulated by Homeobox-containing (Hox) genes.
The upper limb buds appear low on the embryo due to the dominant development of the head and
neck.
The upper limb buds form opposite the caudal cervical segments and lower limb buds form
opposite the lumbar and upper sacral segments.

Limb bud
Each limb bud consists of a mass of mesenchyme derived from the somatic mesoderm, covered by a
layer of ectoderm. At the tip of each limb bud, ectodermal cells form an apical ectodermal ridge, which
promotes growth and development of the limbs in the proximo-distal axis . Fibroblast growth factors and
T-box genes (tbx-4 and tbx-5) from the apical ectodermal ridge activate the mesenchymal cells at the
posterior margin of the limb bud (the zone of polarizing activity). This causes expression of the Sonic
Hedgehog gene, which controls the patterning of the limb along the anterior-posterior axis. Expression
of Wnt7 from the dorsal epidermis of the limb bud and engrailed-1 (EN-1) from the ventral aspect
specifies the dorsal-ventral axis

Week 5

Bones appear during week 5 as mesenchymal condensations in the limb buds

Upper limbs show regional differentiation with developing hand plates.

Week 6

Mesenchymal models of the bones in the limbs undergo chondrification to form hyaline
cartilage.
The clavicle develops by intramembranous ossification and later develops articular cartilages.
The cartilage models form sooner in the upper limb than in the lower limb and in a proximodistal
sequence.

Further differentiation of the limb buds during week 6:

Identifiable elbow and wrists regions are formed.

Hand plates develop ridges, called digital rays and these will become the future thumb and
fingers. At the tip of each digital ray is a portion of the apical ectodermal ridge. It induces
development of the mesenchyme into the primordia of bones. Areas between the rays contain
loose mesenchyme.
Development of the lower limb buds is always slower by a few days.

Week 7

Loose mesenchyme between the digital rays break down and notches appear between the digital
rays in the hand plates.
Digital rays form in the foot plate.
Ossification in the long bones begin by the end of the embryonic period (week 7). The primary
centers are in the diaphyses.
Limb muscles are formed by myogenic precursor cells that migrate into the limb buds and
differentiate into myoblasts. They are derived from the dorsolateral muscle-forming region of the
somites, an area which expresses the muscle-specific genes MyoD and myf-5. Expression of
MyoD results from the influence of activating Wnt proteins and inhibitory BMP-4 protein. The
myoblasts form a muscle mass which divides into a dorsal (extensor) and ventral (flexor)
compartments.

Limb rotation begins:

Originally, the flexor aspect of the limbs is ventral and the extensor aspect is dorsal; the preaxial
border is cranial and the postaxial border is caudal in direction.
The upper limbs rotate 90 degrees on their longitudinal axis. Elbows point posteriorly and
extensor muscles now lie lateral and posterior.
The lower limbs rotate 90 degrees in the opposite direction of rotation of the upper limbs and the
knees face anteriorly. The extensor muscles now lie anteriorly.
The radius in the forearm is homologous to the tibia in the leg, and the ulna is homologous to the
fibula.

Muscles of the limb shift their position during development because of the lateral rotation of the
upper limb and medial rotation of the lower limb.
Muscles forming on the dorsal side of the long bones give rise to extensor and supinator muscles
of the upper limbs and extensor and abductor muscles of the lower limb. They are innervated by
the dorsal branches of the ventral primary rami.
Muscles forming on the ventral side of the long bones become flexor and pronator muscles of the
upper limb and flexor and adductor muscles of the lower limb. They are innervated by the ventral
branches of the ventral primary rami.

Week 8 (Last week of embryonic life;

At the beginning of week 8,

The digits of the hand are short and webbed.

Notches develop between the digital rays of the feet.

At the end of week 8, there are distinct regions in the limbs, with long fingers and distinct toes.
FETAL PERIOD
Weeks 9-12

The fetus has short legs and small thighs at the beginning of week 9.
By the end of week 12, the upper limbs have reached their final relative length but the lower
limbs have not.
Primary ossification centers are present in all long bones.
Order of ossification: Clavicle, femora, etc...

Weeks 34-38

Secondary ossification centers appear in the epiphyses. The first ones to appear are in the distal
end of the femur and the proximal end of the tibia, at the knee joint.
The epiphyseal cartilage plate intervenes between the diaphysis and epiphysis. When it is
replaced around age 25, growth of the bones ends.

A dermatome is the area of skin innervated by a single spinal nerve and its dorsal root ganglion.
Development of the innervation of the limbs

Peripheral nerves grow from the brachial and lumbar plexuses into the mesenchyme of the limb
buds during week 5.
The distribution is segmental, supplying both dorsal and ventral aspects.
As the limbs elongate, the cutaneous distribution follows and an orderly sequence can still be
seen in the adult.
There is no overlap across the axial line.

Development of the blood supply to the limbs

Limb buds are supplied by branches of the intersegmental arteries arising from the aorta.
Initially, a primary axial artery and its branches supply the limb bud and a peripheral marginal sinus
drains it.
In the upper limb,

The primary axial artery becomes the brachial artery in the arm and the common interosseous
artery in the forearm.
o The terminal branches of the brachial artery are the radial and ulnar arteries.
o The terminal branches of the common interosseous arteries are the anterior and posterior
interosseous arteries.
With the formation of the digits the marginal sinus breaks up into the dorsal venous arch. The
final pattern of basilic and cephalic veins and their tributaries then arises.

In the lower limb,

The primary axial artery will form the profunda femoris artery in the thigh, and the anterior and
posterior tibial arteries in the leg.