EARLY EMBRYOLOGY: SOMITE STAGE AND LIMB BUDS

Week 1-2: formation of zygote, implantation and formation of bilaminar embryo (p. 3-4, fig. 1-1). Weeks 3-8: Embryological period (p. 4-5, fig. 1-1). Weeks 9-38: Fetal period (p. 5-6, figs. 1-1 and 1-2).

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 ofparaxial mesoderm. By the end of the 3rd week, the paraxial mesoderm divides into paired bodies called somites, located bilaterally of the neural tube (p. 64, fig. 4-10). 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 (p. 81-89). 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; p. 340, fig. 141).

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 (p. 81, fig. 5.8). 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; p. 84, fig. 5.12). The patterning of the limb development is regulated by Homeoboxcontaining (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 (p. 366, fig. 16-2) 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 (tbx4 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 (p. 371, fig. 16-7) Upper limbs show regional differentiation with developing hand plates (p. 367, fig. 16-3).

Week 6 (p. 354, fig. 14-14; p. 371, fig. 16-7)

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 (p. 367, fig. 16-3):

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 (p. 343, fig. 14-5). 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 andmyf-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 (p. 373, fig. 16-9):

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; p. 372 fig. 16.8) 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 (p. 5-6, fig. 1-1 and 1-2) 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 (p. 343, fig. 14-5). Order of ossification: Clavicle, femora, etc...

Weeks 34-38

Secondary ossification centers appear in the epiphyses (p. 343, fig. 14-5). 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 (p. 373, fig. 16-10). 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 (p. 374, fig. 16-11). 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.

updated 8/25/2008

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Introduction: at day 17, the 3 germ layers are seen (ectoderm above, entoderm below, and mesoderm between). At day 19, the lateral mesodermal plate cleaves and the intraembryonic coelom appears, and differentiation of a somite plate is seen on the side of the neural tube. Metamerization begins at days 20 to 21 with embryonic flexion. Segmentation proceeds caudally, resulting in 42 to 44 pairs of somites by the end of week 5. Each somite develops a myocele. The sclerotome forms on its ventromedial portion and migrates to the notochord where it gives rise to fibroblasts, chondroblasts, and osteoblasts according to location. The rest of the somite, its dorsolateral part, is called the dermomyotome, which forms a dermatome(spreads under the surface ectoderm to form the subcutaneous tissue) and amyotome which forms the skeletal muscles Skeletal muscles are derived from mesenchymal myoblasts which originate in the myotome portion of the dermomyotome. Skeletal muscles may also arise from mesenchyme in the branchial arches and somatic mesoderm A. THE MYOBLASTS elongate, combine to form parallel bundles, and fuse to form multinucleated cells. The central nuclei move to the periphery, and during fetal life myofibrils are seen in the cytoplasm. By month 3, crossstriations are also visible B. THE MYOTOME: most myotome development occurs in the thoracic region by week 5. Each myotome divides into a small dorsal epaxialdivision (epimere) and a larger ventral hypaxial division (hypomere). Each spinal nerve also divides, sending branches to each division, a dorsal primary ramus to the epimere and a ventral primary ramus to the hypomere. Most myotomes migrate to form nonsegmented muscles; some remain segmentally arranged like the somites (e.g., the intercostals of the thorax) 1. Epaxial derivatives: these myoblasts form the extensor muscles of the neck, vertebral column, and lumbar region. Extensors from the caudal sacral and coccygeal myotomes degenerate and become the adult dorsal sacrococcygeal ligament 2. Hypaxial derivatives: myoblasts of cervical myotomes form the scalene, prevertebral, infrahyoid, and geniohyoid muscles. Thoracic myotomes become the lateral and ventral flexors of the vertebral column. Lumbar myotomes become the quadratus lumborum muscl The sacrococcygeal myotomes form the muscles of the pelvic diaphragm, anus, and sex organs C. BRANCHIAL ARCH MUSCLES: myoblasts from the arches migrate to form the muscles of mastication, of facial expression, and muscles of the pharynx and larynx. They are innervated by branchial arch nerves V, VII, IX, and X, respectively

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OCULAR MUSCLES are probably derived from mesenchymal cells around the prochordal plate which gives rise to 3 preoptic myotomes. Groups of myoblasts with cranial nerves III, IV, and VI form the extrinsic muscles of the eyeball E. TONGUE MUSCLES: 4 occipital myotomes are seen first, but the first pair disappears. The last 3 pairs form the tongue muscles, innervated by cranial nerve XII F. LIMB MUSCLES develop in situ from mesenchyme around the developing limb bones. The mesenchyme comes from the somatic layer of the lateral plate mesoderm Visceral (smooth and cardiac) muscle A. SMOOTH MUSCLE forms from splanchnic mesenchyme around the primitive gut and its derivatives. Elsewhere, it forms from local mesenchyme 1. Muscles of the iris (sphincter and dilator pupillae) and the myoepithelial cells of the breast and sweat glands come from mesenchymal cells of ectodermal origin B. CARDIAC (HEART) MUSCLE: forms from splanchnic mesenchyme around the embryonic heart 1. Special bundles of muscle cells develop with few myofibrils. These atypical cells form the Purkinje fibers of the conduction system of the heart Congenital malformations of muscle may be due to muscle failure to develop or a pathologic process affecting the muscle or nerve during embryonic development

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ntroduction: these systems develop from mesoderm. The somites differentiate into 2 parts: cells of the sclerotome give rise to bone, cartilage, and ligaments; those of the dermomyotome give rise to skeletal muscle. The mesodermal cells form mesenchyme (embryonic connective tissue) which can differentiate into fibroblasts, chondroblasts, and osteoblasts. In addition to somite mesenchyme, the splanchnic and somatic mesoderm also can form mesenchyme (some head mesenchyme even arises from neuroectoderm). Most bones first appear as condensations of mesenchymal cells which give rise to hyaline cartilage models that ossify via endochondral ossification. Others develop in mesenchyme by intramembranous bone formation Cartilage histogenesis: cartilage is seen as mesenchymal condensations, at about week 5, where it is to develop. The cells proliferate, round up, and elastic or cartilaginous fibers are deposited in the intercellular substance (matrix). Three types are described: hyaline, fibrocartilage, and elastic cartilage, depending on matrix Bone histogenesis: bone develops in 2 types of preexisting connective tissue, namely, in mesenchyme or cartilage A. INTRAMEMBRANOUS FORMATION develops in mesenchyme 1. The mesenchyme condenses, becomes very vascular, and the cells differentiate into osteoblasts (bone-forming cells) which deposit an intercellular matrix 2. The matrix is calcified to form spicules of spongy bone 3. Some osteoblasts are trapped in the matrix to becomeosteocytes (bone cells) as successive layers of lamellae are deposited by other osteoblasts 4. The spicules thicken, fuse, and form plates of compact bon With internal reorganization, haversian systems develop 5. Between the plates of bone, the intervening bone stays spongy and the mesenchyme forms bone marrow 6. Both osteoblasts and osteoclasts continue to remodel the bone 7. Ossification begins at the end of the embryonic period B. ENDOCHONDRAL OR INTRACARTILAGINOUS OSSIFICATION takes place in a preexisting cartilage model

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In a long bone (, the femur), the primary ossification center is seen in the diaphysis or shaft (between ends of the bone). Here the cartilage increases in size (hypertrophies), the matrix is calcified, and the cells die a. Concurrently, a thin layer of bone is laid down under the perichondrium around the diaphysis and will become theperiosteum b. Vascular connective tissue invades from the periosteum and breaks up the cartilag Some of the mesenchymal cells form hematopoietic cells of the bone marrow and others form osteoblasts which deposit bone matrix on the spicules of calcified cartilag The spicules are remodeled by osteoblasts and osteoc1asts, and the process continues toward both ends of the bones (epiphysis) c. The bone grows in length at the diaphyseoepiphyseal junction where the cartilage cells proliferate by mitosis d. The cartilage cells facing the diaphysis hypertrophy, the matrix is broken up into spicules by the vascular tissue from the marrow, and bone is deposited on the spicules i. Resorption of bone keeps the bone mass relatively constant in length and enlarges the marrow cavity ii. At birth, the diaphyses are largely ossified, but most epiphyses are still cartilaginous

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Limb primordia A. THE LIMB BUD PRIMORDIA appear at the end of week 4 as small elevations of the ventrolateral body wall, but most development occurs in week 6. Early developmental stages for both the upper and lower limbs are identical, except that the arm buds precede those of the leg buds by several days B. THE BUDS are formed by a series of reciprocal inductions of mesoderm and ectoderm 1. The lateral mesoderm of the somatopleure induces a transitory longitudinal thickening of the surface ectoderm, the wolffian crest, a fold which can be seen in front of the somite column. The middle portion of the column disappears rapidly, leaving only 2 nodes at the extremities of the crest which are at the level of the future osseous girdles a. The arm buds develop opposite the caudal cervical segments; the leg buds develop opposite the lumbar and upper sacral segments 2. The ectodermal nodule or apical ectoderm ridge, located on the proximal side of the column, induces the mesenchyme to grow and develop the limbs, in successive waves. There is no apparent contribution from the myotome regions of the somite a. Each bud is thus initially a mass of mesenchyme, of somatic mesodermal origin, covered by ectoderm 3. Innervation of the limbs is an early phenomenon. The upper limb is supplied from the last 6 cervical and first 2 thoracic metameres (brachial plexus); the lower limb from the last 4 lumbar and first 3 sacral metameres (lumbosacral plexus) Limb development 1. The first primordium of the upper limb appears about the 24th day and that of the lower limb at about day 2 The essential basic constituents of the limbs are distinguishable at day 3 2. The distal ends of the limb buds flatten into paddle-shaped hand or foot plates, and the respective digits form at the margins of these plates 3. The limb acquires its distal segment in week Shortly after this, a groove divides the proximal segment, and the limb now consists of its 3 definitive segments. Development of the upper limb is more advanced than that of the lower

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Chondroblasts appear in the precartilaginous matrix which fragments to form the various skeletal parts. Between them, the first joint structures make their appearance toward week 5. As the bones form and limbs elongate, myoblasts aggregate and form the large muscle masses in each limb a. The muscle masses separate into dorsal (extensor) and ventral (flexor) components 6. Early in week 7, the limbs move ventrally, and the developing arms and legs rotate to different degrees and in opposite directions a. Initially, the flexor surface of the limbs is ventral and the extensor surface is dorsal, with the preaxial and postaxial borders being cranial and caudal, respectively b. With rotation, the upper limbs rotate laterally through 90 degrees on their long axes, the elbows come to face posteriorly, and the extensor muscles come to lie on the outer or dorsal aspect of the arm c. With rotation, the lower limbs rotate medially through 90 degrees on their long axes, the knees face forward or ventrolaterally, and the extensor muscles come to lie on the ventral aspect of the legs Dermatome distribution and cutaneous innervation are related to the growth and rotation of the limbs A. A DERMATOME is an area of skin (sensory) supplied by a single spinal nerve and its dorsal root ganglion 1. Peripheral spinal nerves grow from the brachial plexi into the upper limb buds, and lumbosacral plexi grow into the lower limb buds, during week 5 2. The spinal nerves (dermatome nerves) are distributed in segmental bands to supply the dorsal and ventral surfaces of the limb buds a. As the bud elongates, the cutaneous nerves migrate out along the limbs, and although the original dermatomal patterns change with growth, there is still an orderly sequence of nerve distribution b. The limb dermatomes can be traced down the lateral side of the arm and up its medial side, while those of the lower limb can be traced down the ventral side and up the dorsal side c. Autonomic nerves that supply the blood vessels grow out into the limb buds with the spinal nerves B. A CUTANEOUS NERVE supplies an area of the skin that is related to a peripheral nerve. Note that any cutaneous nerve may contain fibers from several individual spinal nerves. Therefore, cutaneous nerve areas and dermatome areas show much overlapping. A cutaneous nerve area is generally broader and wider than an area supplied by only a single spinal (dermatome) nerve

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