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DEVELOPMENT OF LIMB

Dr Anggraini Barlian

Learning objectives






The derivatives of mesoderm
The limb bud formation
The embryological development of upper limb
The embryological development of lower limb
The development of the vertebral column
The limb defects
To describe the vertebral column defects

Derivatives of
mesoderm:

* Paraxial/
somite:
- Myotome
- Dermatome
- Schleretome
* Intermediate
* Lateral
* Head

Classification of mesoderm :
• Notochord: axial mesoderm
in adult substituted by columna vertebrate
schlerotome from somite
• Somite : paraxial mesoderm
– Dermatome  Dermis
– Myotome  skeletal muscle
– Schlerotome  vertebrae, ribs

• Mesoderm intermediate : Urogenital system
• Mesoderm lateral :
– mesoderm splancnic : circulatory system
– mesoderm somatic : limbs

Dermatome  . Cells of ventral & medial walls of somites lose their epithelial arrangement & migrates  notochord Cells at dorsolateral sclerotome migrate limb & body wall musculature.Stages development of somites: A.Myotome continue to extend D. Dorsomedial cells migrate  myotome C. Mesoderm cells are arranged around a small cavity B.

Middorsal 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 .• Expression patterns of genes that regulates somite differentiation. Wnt prot  dorsomedial portion of somite form epaxial muscle & express muscle specific gene Myf5. secreted by notochord & floor plate of neural tube  ventral part of somite form sclerotome & expressed PAX1 control chondrogenesis & vertebral formation. Wnt proteinactivate PAX3 demarcates dermomyotome. Sonic hedgehog (Shh).

LIMB BUD FORMATION .

the mesoderm stops dividing. 3/24/2009 .Limb Bud Formation • The distal tip of the limb bud is covered with a transient structure known as the apical ectodermal ridge (AER). • If the AER is removed. Limb outgrowth is dependent upon interactions between the AER and the underlying mesoderm.

a supernumerary limb results. limb outgrowth involves reciprocal interactions between the AER and underlying mesoderm. • Prospective limb mesoderm grafted under flank ectoderm will promote formation of an AER and a supernumerary limb. • Therefore. the AER regresses and the mesoderm ceases proliferation. • If limb bud mesoderm is removed from an early limb bud. 3/24/2009 .Limb Development • If a supernumerary AER is grafted adjacent to a developing limb.

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Those residing there the longest become the most distal elements. Fates of cells are determined by the length of time they spend in the progress zone.What Controls Establishment of Limb Axes? • The limb has three axes: Proximo-distal. these elements are laid down in a proximal-to-distal direction. The AER does not regulate P-D polarity. Anteriorposterior and Dorsal-ventral. Thus. . • Proximo-distal Removal of the AER at progressively later stages results in a truncated limb with progressively more distal elements.

which is located at the junction between the limb bud and the body wall.Anterior-Posterior Axis This axis is determined by the zone of polarizing activity (ZPA). A diffusible morphogen (retinoic acid?) is released from the ZPA. Grafts of the ZPA to the anterior margin of a host limb bud causes duplication of digits in mirror-image symmetry. 3/24/2009 .

proximal/distal (base of limb / digits) . anterior/posterior (1st to 5th digits) • 2. ventral/dorsal (palm/ dorsum of hand) • 3.Which axis forms first? • 1.

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fibula Radius. ulna Tarsals Carpals Digits Digits – – Metatarsals Metacarpals – Phalanges – Phalanges .Upper Limb Lower • • •• •• •• Pelvis Scapula Femur Humerus Tibia.

ulna Carpals Digits – Metacarpals – Phalanges .Upper Limb • • • • • Scapula Humerus Radius.

000 births (Indiana Hand Center) . at ~ 1 in 100.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.

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 .How do these abnormalities arise? THROUGH IMPROPER DEVELOPMENT!!! The goal of limb development research is to understand how cartilage.

capable of independent survival – Involves: pattern formation. and cellular differentiation . heart.Limb Development is A Part of Organogenesis • The process of organogenesis involves using the basic body plan (organized embryo) to develop specific organs (limbs. specification of positional information. eyes) in specific regions that leads to the fully functional organism. morphogenesis. induction.

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 .

Model System – Development of the Chick Limb I • Chick limbs begin to develop on the 3rd day after laying • The limb develop from limb buds on the body wall of the embryo .

Development of the Chick Limb II .

and other elements gain their positional information . cells in the progress zone proliferate and acquire a positional value •When cells leave the progress zone.Development of the Chick Limb III • As the limb bid grows outward. cartilage may begin to differentiate.

Embryonic Chick Wing (day 10) .

What Directs Limb Development? • The APICAL ECTODERMAL RIDGE (AER) is responsible for the proximo-distal patterning of the limb. which occurs over time through the progress zone • Signals are sent both from the AER to the progress zone. and from the progress zone to the AER .

upper or lower) .e.What Directs Limb Development? (cont’d) • The ZONE OF POLARIZING ACTIVITY (ZPA) is responsible for antero-posterior patterning of the limb • The ECTODERM controls patterning along the dorso-ventral axis • Differences in the activation of genes that control the interpretation of positional information is responsible for development of different limb structures (i.

die on E 5.What Molecules Direct Limb Development? • Sonic Hedgehog (Shh) – The Shh gene encodes inductive signaling molecules that function during embryogenesis – Involved in establishing cell fates – Key signal in patterning of the antero-posterior limb axis – Alx-4 represses Shh expression • Fibroblast Growth Factor 4 (FGF-4) – Member of FGF family – Mesodermal cell stimulator and directs developmental signaling – FGF-4 -/.0 – Transcriptionally inactive in adult tissues .

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. 3/24/2009 . abnormal formation of the mammalian heart) through alterations in gene expression.g..

: syndactyly 3/24/2009 . the fingers would be webbed.Cell Death and Digit Formation Without apoptosis.

.Limb malformation These are clenched hands resulting from camptodactyly (fingers bent over) and clinodactyly (fingers inclined to one side or the other). The feet are turned inward. the appearance of the club feet (varus deformity) seen here. This Deformation from constraint with particular appearance is very oligohydramnios in utero can result in suggestive of trisomy 18.

the foot has rudimentary. though some do.Example of abnormality  absent of hands (+ legs) (amelia/ phocomelia)  caused by thalidomide (anti-vomiting) Syndactyly represents fusion of two or more digits. . Seen above are the digits of the hand fused together. Below. Most of these syndromes do not have a specific genetic defect yet defined. It can be an isolated finding or part of syndromes that define patterns of anomalies.

Loss of mesoderm in the lumbosacral region has resulted in fusion of limb Regulative ability of Limb field . 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. Sirenomelia (caudal dysgenesis) gastrulation disruption.This is sirenomelia.

What are these abnormalities? Polydactyly – the duplication of a digit .

tetracyclin (bone and tooth anomalies) etc Cocaine. hypoxia) Child with unilateral amelia . streptomycin (deafness). limb abnormalities. PCP (CNS. antidepressant (limb deformities). LSD.Caution: aspirin.

MALFORMATION IN CRANIAL DEVELOPMENT .

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VERTEBRA COLUMN DEVELOPMENT .

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Development of vertebrae .