LIMBS Limb Growth and Development

Limbs buds appear in the fourth week. 1. Lateral plate mesoderm forms the bones and connective tissue, 2. somites form the muscles of the limbs. Gene regulation:
The AER gene regulates limb outgrowth, and the ZPA gen controls anteroposterior patterning.

Development of the limb buds in human embryos.

A.

At 5 weeks.

B. At 6 weeks.

C. At 8 weeks.

B.

Hindlimb development lags behind forelimb development by 1 to 2 days.

Longitudinal section through the limb bud showing a core of mesenchyme covered by a layer of ectoderm that thickens at the distal border of the limb to form the apical ectodermal ridge. In humans, this occurs during the fifth week of development.

Schematic of human hands. A. At 48 days. Cell death in the apical ectodermal ridge creates a separate ridge for each digit. B. At 51 days. Cell death in the interdigital spaces produces separation of the digits. C. At 56 days. Digit separation is complete.

Lower extremity of an early 6-week embryo, illustrating the first hyaline cartilage models. B,C. Complete set of cartilage models at the end of the 6th week And the beginning of the 8th week, respectively.

Endochondral bone formation.

A. Mesenchyme cells begin to condense and differentiate into

chondrocytes.

B. Chondrocytes form a cartilaginous model of the prospective bone. C,D. Blood vessels invade the center of the cartilaginous model, bringing osteoblasts (black cells) and restricting proliferating chondrocytic cells to the ends (epiphyses) of the bones.

Molecular regulation of patterning and growth in the limb. A. Limb outgrowth is initiated by FGF10 secreted by lateral plate mesoderm in the limb-forming regions.
Once outgrowth is initiated, the apical ectodermal ridge is induced by bone morphogenetic proteins and restricted in its location by the gene Radical fringe expressed in dorsal ectoderm.

Chondrocytes toward the shaft side (diaphysis) undergo hypertrophy and apoptosis as they mineralize the surrounding matrix. Osteoblasts bind to the mineralized matrix and deposit bone matrices. Later, as blood vessels invade the epiphyses, secondary ossification centers form. Growth of the bones is maintained by
proliferation of chondrocytes in the growth plates

In turn, this expression induces that of SER2 in cells destined to form the apical ectodermal ridge. After the ridge is established, it expresses FGF4 and FGF8 to maintain the progress zone, the rapidly proliferating mesenchyme cells adjacent to the ridge. B. Anteroposterior patterning of the limb is controlled by cells in the zone of polarizing activity at the posterior border. These cells produce retinoic acid (vitamin A),

These cells produce retinoic acid (vitamin A), which initiates expression of sonic hedgehog, regulating patterning. C. The dorsoventral limb axis expressed in the dorsal ectoderm. D. Bone type and shape are regulated by clusters of genes , they are the primary determinants of bone morphology.

Experimental procedure for grafting a new zone of polarizing activity from one limb bud into another using chick embryos. The result is the production of a limb with mirror image duplication of the digits indicating the

role of the zone of polarizing activity in regulating anteroposterior patterning of the limb. Sonic hedgehog protein is

the molecule secreted by the zone of polarizing activity responsible for this regulation.

Bone Age
1)

Clinical Correlates

Radiologists use the appearance of various ossification centers to determine whether a child has reached his or her proper maturation age. Useful information about bone age is obtained from ossification studies in the hands and wrists of children.
Prenatal analysis of fetal bones by ultrasonography provides information about fetal growth and gestational age.

2)

3)

Limb Defects
Limb malformations occur in approximately 6\10,000 live births, with 3.4 \10,000 affecting the upper limbs and 1.1 \10,000 affecting the lower limbs. These defects are often associated with other birth defects involving the 1) craniofacial, 2) cardiac, 3) and genitourinary systems.

Abnormalities of the limbs

1) 2) 3)

partial absence of one or more of the extremities (meromelia) complete absence (amelia) of one or more of the extremities Sometimes the long bones are absent, and rudimentary hands and feet are attached to the trunk by small, irregularly shaped bones (phocomelia, a form of meromelia)

4)

Sometimes all segments of the extremities are present but abnormally short (micromelia).

Child with unilateral amelia and multiple defects of the left upper limb.
B. Patient with a form of meromelia called phocomelia. The hands are attached to the trunk by irregularly shaped bones.

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mainly hereditary, cases of teratogen-induced limb defects have been documented.. Many mothers of these infants had taken thalidomide, a drug widely used as a sleeping pill and antinauseant. thalidomide causes a characteristic syndrome of malformations consisting of 1. absence or gross deformities of the long bones, 2. intestinal atresia. 3. cardiac anomalies. Because the drug is now being used to treat AIDS and cancer patients, there is concern that its return will result in a new wave of limb defects. Studies indicate that the most sensitive period for teratogen-induced limb malformations is the fourth and fifth weeks of development.

A different category of limb defects involves the digits.

Sometimes the digits are shortened(brachydactyly); If two or more fingers or toes are fused, it is called syndactyly Normally, mesenchyme between prospective digits in hand- and footplates is removed by cell death (apoptosis). In one \2,000 births this process fails, and the result is fusion between two or more digits.

The presence of extra fingers or toes is called polydactylyThe extra digits frequently lack proper muscle connections. Abnormalities involving polydactyly are usually bilateral, whereas absence of a digit (ectrodactyly), such as a thumb, usually occurs unilaterally.

Digital defects.
A. Brachydactyly, short digits . B. Syndactyly, fused digits. C. Polydactyly, extra digits. D. Cleft foot, lobster claw deformity. Any of these defects may involve either the hands or feet or both.

A number of gene mutations have been identified that affect the limbs and sometimes other structures
Result in 1-hand-foot-genital syndrome 2-Holt-Oram syndrome. 3- Osteogenesis imperfecta 5-Clubfoot . 6-Congenital absence or deficiency of the radius . 7-craniosynostosis-radial aplasia syndrome 8-Congenital hip dislocation

VERTEBRAE AND THE VERTEBRAL COLUMN

The vertebral column and ribs develop from the sclerotome compartments of the somites, and the sternum is derived from mesoderm in the ventral body wall.

A definitive vertebra
is formed by condensation of the caudal half of one sclerotome and fusion with the cranial half of the subjacent sclerotome

The many abnormalities of the skeletal system include

1. 2. 3.

vertebral (spina bifida), cranial (cranioschisis and craniosynostosis), facial (cleft palate) defects.

Major malformations of the limbs are rare, but defects of the radius and digits are often associated with other abnormalities (syndromes).

As the vertebrae form, two primary curves of the spine are established:
1. 2.

the thoracic curvature The sacral curvature.

Later, two secondary curves are established:

1- cervical curvature , as the child learns to hold up his or her head 2- the lumbar curvature ,which forms when the child learns to walk.

Scoliosis

Vertebral Defects

(lateral curving of the spine). When two successive vertebrae fuse asymmetrically or have half a vertebra missing, a cause of scoliosis

(spina bifida),
One of the most serious vertebral defects is the result of imperfect fusion or nonunion of the vertebral arches. Such an abnormality, known as cleft vertebra (spina bifida), (1-spina bifida occulta). involve only the bony vertebral arches, leaving the spinal cord intact. the bony defect is covered by skin, and no neurological deficits occur (spina bifida occulta).

2-spina bifida cystica

A more severe abnormality in which the neural tube fails to close, vertebral arches fail to form, and neural tissue is exposed, Any neurological deficits depend on the level and extent of the lesion This defect, 1. which occurs in one per 1,000 births, 2. may be prevented, in many cases, by providing mothers with folic acid prior to conception. 3. Spina bifida can be detected prenatally by ultrasound, and if neural tissue is exposed, amniocentesis can detect elevated levels of fetoprotein in the amniotic fluid.

RIBS:
1-The bony portion of each rib is derived

from sclerotome cells that remain in the paraxial mesoderm and that grow out from the costal processes of thoracic vertebrae.

2-Costal cartilages

are formed by sclerotome cells that migrate across the lateral somitic frontier into the adjacent lateral plate mesoderm

The sternum
develops independently in lateral plate mesoderm in the ventral body wall.

Rib Defects
Clinical Correlates
.

Cervical ribs
occur in approximately 1% of the population and are usually attached to the seventh cervical vertebra. Because of its location, this type of rib may impinge on the brachial plexus or the subclavian artery, resulting in varying degrees of anesthesia in the limb.

Defects of the Sternum

1-Cleft sternum
is a very rare defect and may be complete or located at either end of the sternum. Thoracic organs are covered only by skin and soft tissue. The defect arises when the sternal bands fail to grow together in the midline.
2- Hypoplastic ossification centers

3- premature fusion of sternal segments

also occur particularly in infants with congenital heart defects (20% to 50%)

4- Multiple manubrial ossification centers occur in 6% to 20%

of all children but are especially common in those with Down syndrome

Pectus excavatum is the term for a depressed sternum that is sunken posteriorly. Pectus carinatum refers to a flattening of the chest bilaterally with an anteriorly projecting sternum.
. Both defects may result from abnormalities

of ventral body wall closure or formation of the costal cartilages and sternum