Vertebral Column During the fourth week of development, cells of the sclerotomes shift their position to surround both

the spinal cord and the notochord (Fig. 8.1). This mesenchymal column retains traces of its segmental origin, as the sclerotomic blocks are separated by less dense areas containing intersegmental arteries (Fig. 8.21A). During further development the caudal portion of each sclerotome segment proliferates extensively and condenses (Fig. 8.21 B). This proliferation is so extensive that it proceeds into the subjacent intersegmental tissue and binds the caudal half of one sclerotome to the cephalic half of the subjacent sclerotome (arrows in Fig. 8.21,A and B). Hence, by incorporation of the intersegmental tissue into the precartilaginous vertebral body (Fig. 8.21B), the body of the

ABC
Figure 8.21 Formation of the vertebral column at various stages of development. A. At the fourth week of development, sclerotomic segments are separated by less dense intersegmental tissue. Note the position of the myotomes, intersegmental arteries, and segmental nerves. B. Condensation and proliferation of the caudal half of one sclerotome proceed into the intersegmental mesenchyme and cranial half of the subjacent sclerotome (arrows in A and B). Note the appearance of the intervertebral discs. C. Precartilaginous vertebral bodies are formed by the upper and lower halves of two successive sclerotomes and the intersegmental tissue. Myotomes bridge the intervertebral discs and, therefore, can move the vertebral column. 194 Part Two: Special Embryology

vertebra becomes intersegmental. Patterning of the shapes of the different vertebra is regulated by HOX genes. Mesenchymal cells between cephalic and caudal parts of the original sclerotome segment do not proliferate but fill the space between two precartilaginous vertebral bodies. In this way they contribute to formation of the intervertebral disc (Fig. 8.21B). Although the notochord regresses entirely in the region of the vertebral bodies, it persists and enlarges in the region of the intervertebral disc. Here it contributes to the nucleus pulposus, which is later surrounded by circular fibers of the annulus fibrosus. Combined, these two structures form the intervertebral disc (Fig. 8.21C ). Rearrangement of sclerotomes into definitive vertebrae causes the myotomes to bridge the intervertebral discs, and this alteration gives them the capacity to move the spine (Fig. 8.21C ). For the same reason, intersegmental arteries, at first lying between the sclerotomes, now pass midway over the vertebral bodies. Spinal nerves, however, come to lie near the intervertebral discs and leave the vertebral column through the intervertebral foramina.

CLINICALCORRELATES

Vertebral Defects The process of formation and rearrangement of segmental sclerotomes into definitive vertebrae is complicated, and it is fairly common to have two successive vertebrae fuse asymmetrically or have half a vertebramissing, a cause of scoliosis (lateral curving of the spine). Also, the number of vertebrae is frequently more or less than the norm. A typical example of these abnormalities is found in patients with Klippel-Feil anomaly. These patients have fewer than normal cervical vertebrae, and often other vertebrae are fused or abnormal in shape. This anomaly is usually associated with other abnormalities. 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), may involve only the bony vertebral arches, leaving the spinal cord intact. In these cases the bony defect is covered by skin, and no neurological deficits occur (spina bifida occulta). A more severe abnormality is spina bifida cystica, 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, which occurs in 1/1000 births, may be prevented, in many cases, by providing mothers with folic acid prior to conception. Spina bifida can be detected prenatally by ultrasound (Fig. 8.22), and if neural tissue is exposed, amniocentesis can detect elevated levels of -fetoprotein in the amniotic fluid Summary The skeletal system develops from mesenchyme, which is derived from the mesodermal germ layer and from neural crest. Some bones, such as the flat bones of the skull, undergo membranous ossification; that is, mesenchyme cells are directly transformed into osteoblasts (Fig. 8.2). In most bones, such as the long bones of the limbs, mesenchyme condenses and forms hyaline cartilage models of bones (Fig. 8.15). Ossification centers appear in these cartilage models, and the bone gradually ossifies by endochondral ossification. The skull consists of the neurocranium and viscerocranium (face). The neurocranium includes a membranous portion, which forms the cranial vault, and a cartilaginous portion (chondrocranium), which forms the base of the skull. Neural crest cells form the face, most of the cranial vault, and the prechordal part of the chondrocranium (the part that lies rostral to the notochord). Paraxial mesoderm forms the remainder of the skull. Limbs form as buds along the body wall that appear in the fourth week. Lateral plate mesoderm forms the bones and connective tissue, while muscle cells migrate to the limbs from the somites. The AER regulates limb outgrowth, and the ZPA controls anteroposterior patterning. Many of the genes that regulate limb growth and patterning have been defined (see Fig. 8.16). 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 (Fig. 8.21). The many abnormalities of the skeletal system include vertebral (spina bifida), cranial (cranioschisis and craniosynostosis), and 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 notochord and neural tube form, the intraembryonic mesoderm lateral to these structures thickens to form two longitudinal columns of paraxial mesoderm (Fig. 14-1A and B). Toward the end of the third week, these dorsolateral columns located in the trunk become segmented into blocks of mesoderm-the somites (see Fig. 14-1C). Externally the somites appear as beadlike elevations along the dorsolateral surface of the embryo (see Chapter 5). Each somite differentiates into two parts (see Fig. 14-1D and E):

The ventromedial part is the sclerotome; its cells form the vertebrae and ribs. The dorsolateral part is the dermomyotome; cells from its myotome region form myoblasts (primordial muscle cells), and those from its dermatome region form the dermis (fibroblasts).

DEVELOPMENT OF THE AXIAL SKELETON The axial skeleton is composed of the cranium (skull), vertebral column, ribs, and sternum. During the fourth week, cells in the sclerotomes now surround the neural tube (primordium of spinal cord) and the notochord, the structure about which the primordia of the vertebrae develop. This positional change of the sclerotomal cells is effected by differential growth of the surrounding structures and not by active migration of sclerotomal cells. The Pax-1 gene, which is expressed in all prospective sclerotomal cells of epithelial somites in chick and mouse embryos, seems to play an essential role in the development of the vertebral column. Development of the Vertebral Column During the precartilaginous or mesenchymal stage, mesenchymal cells from the sclerotomes are found in three main areas (Fig. 14-7A): around the notochord, surrounding the neural tube, and in the body wall. In a frontal section of a 4-week embryo, the sclerotomes appear as paired condensations of mesenchymal cells around the notochord (see Fig. 14-7B). Each sclerotome consists of loosely arranged cells cranially and densely packed cells caudally. Some densely packed cells move cranially, opposite the center of the myotome, where they form the intervertebral (IV) disc (see Fig. 14-7C and D). The remaining densely packed cells fuse with the loosely arranged cells of the immediately caudal sclerotome to form the mesenchymal centrum, the primordium of the body of a vertebra. Thus, each centrum develops from two adjacent sclerotomes and becomes an intersegmental structure. The nerves now lie in close relationship to the intervertebral discs, and the intersegmental arteries lie on each side of the vertebral bodies. In the thorax, the dorsal intersegmental arteries become the intercostal arteries.

Figure 14-7 A, Transverse section through a 4-week embryo. The arrows indicate the dorsal growth of the neural tube and the simultaneous dorsolateral movement of the somite remnant, leaving behind a trail of sclerotomal cells. B, Diagrammatic frontal section of this embryo showing that the condensation of sclerotomal cells around the notochord consists of a cranial area of loosely packed cells and a caudal area of densely packed cells. C, Transverse section through a 5-week embryo showing the condensation of sclerotomal cells around the notochord and neural tube, which forms a mesenchymal vertebra. D, Diagrammatic frontal section illustrating that the vertebral body forms from the cranial and caudal halves of two successive sclerotomal masses. The intersegmental arteries now cross the bodies of the vertebrae, and the spinal nerves lie between the vertebrae. The notochord is degenerating except in the region of the intervertebral disc, where it forms the nucleus pulposus. The notochord degenerates and disappears where it is surrounded by the developing vertebral bodies. Between the vertebrae, the notochord expands to form the gelatinous center of the intervertebral disc-the nucleus pulposus (see Fig. 14-7D). This nucleus is later surrounded by circularly arranged fibers that form the anulus fibrosus. The nucleus pulposus and anulus fibrosus together constitute the intervertebral disc. The mesenchymal cells, surrounding the neural tube, form the neural arch (Fig. 14-7C). The mesenchymal cells in the body wall form the costal processes that form ribs in the thoracic region. Chordoma Remnants of the notochord may persist and give rise to a chordoma. Approximately one third of these slow-growing malignant tumors occur at the base of the cranium and extend to the nasopharynx. Cordomas infiltrate bone and are difficult to remove. Few patients survive longer than 5 years. Chordomas also develop in the lumbosacral region.

Cartilaginous Stage of Vertebral Development During the sixth week, chondrification centers appear in each mesenchymal vertebra (Fig. 14-8A and B). The two centers in each centrum fuse at the end of the embryonic period to form a cartilaginous centrum. Concomitantly, the centers in the neural arches fuse with each other and the centrum. The spinous and transverse processes develop from extensions of chondrification centers in the neural arch. Chondrification spreads until a cartilaginous vertebral column is formed.

Figure 14-8 Stages of vertebral development. A, Mesenchymal vertebra at 5 weeks. B, Chondrification centers in a mesenchymal vertebra at 6 weeks. The neural arch is the primordium of the vertebral arch. C, Primary ossification centers in a cartilaginous vertebra at 7 weeks. D, Thoracic vertebra at birth consisting of three bony parts. Note the cartilage between the halves of the vertebral arch and between the arch and the centrum (neurocentral joint). E and F, Two views of a typical thoracic vertebra at puberty showing the location of the secondary centers of ossification. Ossification of typical vertebrae begins during the embryonic period and usually ends by the 25th year. There are two primary ossification centers, ventral and dorsal, for the centrum (see Fig. 14-8C). These primary ossification centers soon fuse to form one center. Three primary centers are present by the end of the embryonic period: one in the centrum and one in each half of the neural arch. Ossification becomes evident in the neural arches during the eighth week. At birth, each vertebra consists of three bony parts connected by cartilage (see Fig. 14-8D). The bony halves of the vertebral arch usually fuse during the first 3 to 5 years. The arches first unite in the lumbar region, and union progresses cranially. The vertebral arch articulates with the centrum at cartilaginous neurocentral joints, which permit the vertebral arches to grow as the spinal cord enlarges. These joints disappear when the vertebral arch fuses with the centrum during the third to sixth years. Five secondary ossification centers appear in the vertebrae after puberty:

One for the tip of the spinous process One for the tip of each transverse process

Two anular epiphyses, one on the superior and one on the inferior rim of the vertebral body (see Fig. 14-8E and F).

The vertebral body is a composite of the anular epiphyses and the mass of bone between them. The vertebral body includes the centrum, parts of the vertebral arch, and the facets for the heads of the ribs. All secondary centers unite with the rest of the vertebra at approximately 25 years of age. Exceptions to the typical ossification of vertebrae occur in the atlas or C1 vertebra, axis or C2 vertebra, C7 vertebra, lumbar vertebrae, sacrum, and coccyx. Minor developmental anomalies of the vertebrae are common, but in most cases are of little clinical importance. Variation in the Number of Vertebrae Most people have 7 cervical, 12 thoracic, 5 lumbar, and 5 sacral vertebrae. A few have one or two additional vertebrae or one fewer. To determine the number of vertebrae, it is necessary to examine the entire vertebral column because an apparent extra (or absent) vertebra in one segment of the column may be compensated for by an absent (or extra) vertebra in an adjacent segment; for example, 11 thoracic-type vertebrae with 6 lumbar-type vertebrae.