Chapter 3 Development of the Nervous System
The entire nervous system and the special sense organs originate from three sources, each derived from specific cell populations of the early epiblast termed neural ectoderm. The first source to be clearly delineated is the neural plate, which gives rise to the central nervous system (CNS), the somatic motor nerves and the preganglionic autonomic nerves. The second source is from cells at the perimeter of the neural plate, which remove themselves by epithelial–mesenchymal transition from the plate just prior to its fusion as a neural tube. These are the neural crest cells, and they form nearly all the peripheral nervous system (PNS), including the somatic sensory nerves, somatic and autonomic ganglia, postganglionic autonomic nerves and adrenal and chromaffin cells. They also give rise to significant mesenchymal populations in the head. The third source is from ectodermal placodes, a group of cells that originate at the edge of the neural plate but remain in the surface ectoderm after neural tube formation, undergoing epithelial–mesenchymal transformation after the neural crest cells have started their migration. Ectodermal placodes contribute to the cranial sensory ganglia, the hypophysis, the inner ear and, by a non-neuronal contribution, the lens of the eye.
Neurulation
Primary neurulation begins at stage 9 (Fig. 3.1). Although the process is continuous spatially and temporally, it has been envisaged as four stages. It begins with local elongation of the ectoderm cells in a midline zone of the disc and their reorganization into a pseudostratified epithelium, the neural plate. This is followed by reshaping of the neural plate and bending of the plate into a neural groove. The latter is closed to form a neural tube bidirectionally from the midportion to its cranial and caudal ends. A continuous surface ectoderm forms dorsal to the tube.
Early Vesicles and Flexures of the Neural Tube
Prior to closure of the neural tube, the neural folds become considerably expanded in the head region—the first indication of a brain. After the rostral neuropore closes, these regional expansions form three primary cerebral vesicles (Fig. 3.2). The term ‘vesicle’ may be a misnomer, because it suggests an exaggerated view of these localized accelerations of growth in the wall of the brain. The bulging is not initially marked, and the vesicles are more like gently fusiform tubes. The three regions are the prosencephalon (forebrain), mesencephalon (midbrain) and rhombencephalon (hindbrain), the last being continuous caudally with the spinal cord. As a result of unequal growth of their different regions, the prosencephalon and rhombencephalon enlarge more than the mesencephalon and can be subdivided. The prosencephalon gives rise to a midline diencephalon and bilateral telencephalon; the rhombencephalon gives rise to the metencephalon and myelencephalon. A summary of the derivatives of the cerebral vesicles is given in Table 3.1.
Rhombencephalon (or hindbrain) | |
1. Myelencephalon | Medulla oblongata |
Caudal part of the fourth ventricle | |
Inferior cerebellar peduncles | |
2. Metencephalon | Pons |
Cerebellum | |
Middle part of the fourth ventricle | |
Middle cerebellar peduncles | |
3. Isthmus rhombencephali | Superior medullary velum |
Superior cerebellar peduncles | |
Rostral part of the fourth ventricle | |
Mesencephalon (or midbrain) | Cerebral peduncles |
Tegmentum | |
Tectum | |
Aqueduct | |
Prosencephalon (or forebrain) | |
1. Diencephalon | Thalamus |
Metathalamus | |
Subthalamus | |
Epithalamus | |
Caudal part of the hypothalamus | |
Caudal part of the third ventricle | |
2. Telencephalon | Rostral part of the hypothalamus |
Rostral part of the third ventricle | |
Cerebral hemispheres | |
Lateral ventricles | |
Cortex (archaeocortex, palaeocortex, neocortex) | |
Corpus striatum |
Elongation of the brain occurs at the same time as the appearance of three flexures that are also developing prior to closure of the neural tube; two are concave ventrally, and one is concave dorsally. During stages 13 and 14 the brain bends at the mesencephalon (mesencephalic flexure), so the prosencephalon bends in a ventral direction around the cephalic end of the notochord and foregut until its floor lies almost parallel to that of the rhombencephalon (see Fig. 3.2). A bend also appears at the junction of the rhombencephalon and spinal cord (cervical flexure). This increases from the fifth to the end of the seventh week, by which time the rhombencephalon forms nearly a right angle to the spinal cord. However, after the seventh week, extension of the head takes place, and the cervical flexure diminishes and eventually disappears. The third bend, the pontine flexure, is directed ventrally between the metencephalon and myelencephalon. It does not substantially affect the outline of the head. In this region the roof plate thins until it is composed of only a single layer of cells and pia mater, the tela choroidea. The flexure of the neural tube at this point produces a rhombic shape in the roof that later forms the medullary velum.
In addition to these gross divisions, a number of ridges and depressions are present transiently on the inner surface of the brain. Prominent among these are the serial bulges that appear very early in the rhombencephalon, before the main flexures of the neural tube develop (Fig. 3.3). These bulges are termed rhombomeres.
Early Cellular Arrangement of the Neural Tube
Histologically, the early neural tube is composed of a pseudostratified neuroepithelium. It extends from the inner aspect of the tube to the outer limiting basal lamina and surrounding neural crest, which will form the pia mater. The epithelium contains stem cells that will give rise to populations of neuroblasts and glioblasts. A population of radial glia differentiates very early and provides a scaffold for later cells to follow. As development proceeds, three zones or layers develop (Figs. 3.4–3.6): an internal ventricular zone (variously termed the germinal, primitive ependymal or matrix layer), in which mitosis occurs and that contains the nucleated parts of the columnar cells and rounded cells undergoing mitosis; a middle mantle zone (also termed the intermediate zone), which contains the migrant cells from the divisions occurring in the ventricular zone; and an outer marginal zone, which initially consists of the external cytoplasmic processes of the radial glia and the neuroepithelial stem cells. The last is soon invaded by tracts of axonal processes that grow from neuroblasts developing in the mantle zone, together with varieties of non-neuronal cells (glial cells and later vascular endothelium and perivascular mesenchyme).
At first the neural tube caudal to the brain is oval in transverse section, and its lumen is narrow and slit-like (see Fig. 3.4). The original floor plate and the dorsal site of fusion of the tube initially contain non-neural cells. With cellular proliferation, the lateral walls thicken and the lumen, now the central canal, widens in its dorsal part and is somewhat diamond shaped on cross-section (see Fig. 3.6). Widening of the canal is associated with the development of a longitudinal sulcus limitans on each side. This divides the ventricular and mantle (intermediate) zones in each lateral wall into a ventrolateral lamina or basal plate and a dorsolateral lamina or alar plate. This separation indicates a fundamental functional difference.
Throughout the neural tube there is a generic pattern in the position of the neurones, specified by the juxtaposition of the notochord to the neural tube. Lateral or dorsal grafting of a notochord results in the induction of a floor plate overlying the grafted notochord and the induction of ectopic dorsal motor neurones. Similarly, lateral or dorsal grafts of a floor plate also result in the induction of a new floor plate overlying the graft and the induction of ectopic dorsal motor neurones. Removal of the notochord results in the elimination of the floor plate and motor neurones and the differentiation of dorsal cell types in the ventral region of the cord (Fig. 3.7).
Fig. 3.7 A–D, Successive stages in the development of the neural tube and spinal cord. A, The neural plate consists of epithelial cells. Cells in the midline of the neural plate are contacted directly by the notochord. More lateral regions of the neural plate overlie the paraxial mesenchyme (not shown). B, During neurulation, the neural plate bends at its midline, which elevates the lateral edges of the plate as the neural folds. Contact between the midline of the neural plate and the notochord is maintained at this stage. C, The neural tube is formed when the dorsal tips of the neural folds fuse. Cells in the region of fusion form the roof plate, which is a specialized group of dorsal midline cells. D, Cells at the ventral midline of the neural tube retain proximity to the notochord and differentiate into the floor plate. After neural tube closure, neuroepithelial cells continue to proliferate and eventually differentiate into defined classes of neurones at different dorsoventral positions within the spinal cord. For example, sensory relay, commissural and other classes of dorsal neurones (D) differentiate near the roof plate (R), and motor neurones (M) differentiate ventrally near the floor plate (F), which by this time is no longer in contact with the notochord (N). E–H, Summary of the results of experiments in chick embryos in which the notochord or floor plate is grafted to the dorsal midline of the neural tube or the notochord is removed before neural tube closure. E, The normal condition, showing the ventral location of motor neurones (M) and the dorsal location of sensory relay neurones (D). F, Dorsal grafts of a notochord result in induction of a floor plate in the dorsal midline and ectopic dorsal motor neurones (M). G, Dorsal grafts of a floor plate induce a new floor plate in the dorsal midline and ectopic dorsal motor neurones (M). H, Removal of the notochord results in the elimination of the floor plate and motor neurones and the expression of dorsal cells types (D) in the ventral region of the spinal cord.
(After Jessell, Dodd. 1992. WB Saunders.)
Failure of Neurulation
Failure of neurulation produces the conditions of craniorachischisis totalis (the entire neural tube is unfused in the dorsal midline), cranioschisis or anencephaly (the neural tube is fused dorsally to form the spinal cord but is not fused dorsally in the brain) and spina bifida (local regions of the spinal neural tube are unfused, or there is failure of formation of the vertebral neural arches) (Fig. 3.8). Anencephalic fetuses display severe disturbances in the shape, position and ossification of the basichondrocranium and in the course of the intracranial notochord (Fig. 3.9).
Neural Crest
The neuronal populations of the early epiblast are arranged in the medial region of the embryonic disc as the neural plate. Laterally, neural folds or crests indicate the transitional region between neural and surface ectoderm. Along most of the neuraxis the cells at the tips of the neural folds undergo an epithelial–mesenchymal transformation. They acquire migratory properties and leave the epithelium just prior to its fusion with the contralateral fold in the dorsal midline. The migratory cells so formed are collectively termed the neural crest (Brown, Keynes and Lumsden 2001). Cells within the rostral prosencephalic neural fold and smaller populations of cells in bilateral sites along the early brain do not form migratory neural crest cells but remain within the surface epithelium as ectodermal placodes.
Neural crest populations arise from the neural folds as primary neurulation proceeds and simultaneously progress rostrally and caudally. Crest cells migrate from the neural folds of the brain prior to tube closure. Caudally, from somite 27, secondary neurulation processes produce the most caudal neural crest. Two distinct populations of neural crest cells are formed: a neuronal population produced throughout the brain and spinal cord that gives rise to sensory and autonomic neurones and glia, and a non-neuronal mesenchymal population that arises only from the brain (Figs. 3.10, 3.11). Melanocytes develop from a subpopulation of neural crest cells derived from both the head and the trunk. They form one of the three pigment cell types (the others being retinal pigment epithelium and pigment cells of the pineal organ, both of which originate from the diencephalon).
In the trunk the migration patterns of neural crest cells are channelled by the somites. As the crest cells move laterally and ventrally they can pass between the somites and within the rostral sclerotomal half of each somite, but they cannot penetrate the caudal moiety of the sclerotomal mesenchyme. Thus the segmental distribution of the spinal and sympathetic ganglia is imposed on the neural crest cells by a prepattern that exists within the somitic paraxial mesenchyme (Fig. 3.12).
Rostral to the otic vesicle, neural crest cells arise from specific regions of the brain. Within the rhombencephalon a number of transverse subdivisions perpendicular to the long axis of the brain can be seen early in development. These are termed rhombomeres (neuromeres) to note their segmental arrangement (Muller and O’Rahilly 1997). At stage 9, six primary rhombomeres can be seen. Up to 16 secondary segments can be identified at stage 14. Eight main rhombomeres are recognized extending from the midbrain–hindbrain boundary rostrally to the spinal cord caudally (see Fig. 3.3). Rhombomeres 8 and 7 give rise to crest cells that migrate into the fourth and sixth pharyngeal arches; rhombomere 6 crest invades the third pharyngeal arch. Rhombomere 4 crest migrates into the second arch, whereas rhombomeres 5 and 3 give rise to a very small number of neural crest cells that migrate rostrally and caudally to enter the adjacent even-numbered neighbours. Rhombomeres 1 and 2 produce crest that invades the first pharyngeal arch. In each case mesenchymal populations and the sensory and autonomic ganglia are formed from the crest cells.
Ectodermal Placodes
Prior to neural tube closure, the elevating neural folds contain two distinctive neuronal populations. The larger population of neural crest cells migrates from the neural epithelium prior to neural tube fusion. A smaller population of neuroepithelial cells becomes incorporated into the surface ectoderm after neural tube closure. These areas of neuroepithelium within the surface ectoderm are termed ectodermal placodes. Although the majority of the ectodermal placodes form nervous tissue, non-neurogenic placodes also occur (Begbie and Graham 2001). After an appropriate inductive stimulus, local clusters of placodal cells remove themselves from the surrounding surface ectoderm either by epithelial–mesenchymal transition or by invagination of the whole placodal region to form a vesicle beneath the remaining surface ectoderm. Neurogenic placodes undergo both processes. Paired non-neurogenic placodes invaginate to form the lens vesicles under the inductive influence of the optic vesicles.
The neural folds meet in the rostral midline adjacent to the buccopharyngeal membrane. This rostral neural fold does not generate neural crest but gives rise to the hypophysial placode (i.e. the future Rathke’s pouch), which remains within the surface ectoderm directly rostral to the buccopharyngeal membrane. The rostral neural fold also gives rise to the olfactory placodes (which remain as paired, laterally placed placodes) and to the epithelium of the nasal cavity (see Fig. 3.11).
Further caudally, similar neurogenic placodes can be identified and divided into three categories: ventrolateral or epibranchial, dorsolateral and intermediate (Fig. 3.13). The epibranchial placodes appear in the surface ectoderm immediately dorsal to the area of pharyngeal (branchial) cleft formation. The first epibranchial placode is located at the level of the first pharyngeal groove and contributes cells to the distal (geniculate) ganglion of the facial nerve; the second and third epibranchial placodes contribute cells to the distal ganglia of the glossopharyngeal (petrosal) and vagus (nodose) nerves, respectively. Generally these placodes thicken, and cells begin to detach from the epithelium soon after the pharyngeal pouches have contacted the overlying ectoderm. Concurrently the neural crest cells reach and move beyond these lateral extensions of the pharynx. Cells budding off placodes show signs of early differentiation into neurones, including the formation of neurites. Epibranchial placodes may have their origins in the neurones that innervate the taste buds in fishes.
Intermediate between the epibranchial and dorsolateral placodes are the profundal and trigeminal placodes, which fuse in humans to form a single entity. Prospective neuroblasts migrate from foci dispersed throughout the surface ectoderm lateral and ventrolateral to the caudal mesencephalon and metencephalon to contribute to the distal portions of the trigeminal ganglia.
Pituitary Gland (Hypophysis Cerebri)
The most rostral portion of the neural plate, which will form the hypothalamus, is in contact rostrally with the future adenohypophysis, in the rostral neural ridge, and caudally with the neurohypophysis, in the floor of the neural plate (see Fig. 3.11). After neurulation the cells of the anterior neural ridge remain in the surface ectoderm and form the hypophysial placode, which is in close apposition and adherent to the overlying prosencephalon.
Neural crest mesenchyme later moves between the prosencephalon and surface ectoderm, except at the region of the placode. Before rupture of the buccopharyngeal membrane, proliferation of the periplacodal mesenchyme results in the placode forming the roof and walls of a saccular depression. This hypophysial recess (Rathke’s pouch; Figs. 3.14, 3.15) is the rudiment of the adenohypophysis. It lies immediately ventral to the dorsal border of the buccopharyngeal membrane, extending in front of the rostral tip of the notochord and retaining contact with the ventral surface of the prosencephalon. It is constricted by continued proliferation of the surrounding mesenchyme to form a closed vesicle, but it remains connected for a time to the ectoderm of the stomodeum by a solid cord of cells that can be traced down the posterior edge of the nasal septum. Masses of epithelial cells form mainly on each side and in the ventral wall of the vesicle, and development of the adenohypophysis progresses by the ingrowth of a mesenchymal stroma. Differentiation of epithelial cells into stem cells and three differentiating types is apparent during the early months of fetal development. It has been suggested that different types of cells arise in succession and that they may be derived in varying proportions from different parts of the hypophysial recess. A craniopharyngeal canal, which sometimes runs from the anterior part of the hypophysial fossa of the sphenoid to the exterior of the skull, often marks the original position of the hypophysial recess. Traces of the stomodeal end of the recess are usually present at the junction of the septum of the nose and the palate. Some claim that the craniopharyngeal canal itself is a secondary formation caused by the growth of blood vessels and that it is unconnected to the stalk of the anterior lobe.
Just caudal to, but in contact with, the adenohypophysial recess, a hollow diverticulum elongates toward the stomodeum from the floor of the neural plate just caudal to the hypothalamus (see Fig. 3.15B); this region of neural outgrowth is the neurohypophysis. It forms an infundibular sac, the walls of which increase in thickness until the contained cavity is obliterated except at its upper end, where it persists as the infundibular recess of the third ventricle. The neurohypophysis becomes invested by the adenohypophysis, which extends dorsally on each side of it. The adenohypophysis gives off two processes from its ventral wall that grow along the infundibulum and fuse to surround it, coming into contact with the tuber cinereum and forming the tuberal portion of the hypophysis. The original cavity of Rathke’s pouch remains first as a cleft and later as scattered vesicles; it can be identified readily in sagittal sections through the mature gland. The dorsal wall of Rathke’s pouch, which remains thin, fuses with the adjoining part of the neurohypophysis as the pars intermedia.
At birth the hypophysis is about one-sixth the weight of the adult gland; it increases to become about one-half the weight of the adult gland at 7 years and attains adult weight at puberty. Throughout postnatal life the gland is both larger and heavier in females.
Neuroglia
The ventricular zone lining the early central canal of the spinal cord and the cavities of the brain gives rise to neurones and glial cells (see Figs. 3.4, 3.5). One specialized form of glial cell is the radial glial cell, whose radial processes extend both outward, to form the outer limiting membrane deep to the pia mater, and inward, to form the inner limiting membrane around the central cavity. The geometry of these cells may provide contact guidance paths for cell migrations, both neuroblastic and glioblastic. A secondary radial glial scaffold is formed in the late-developing cerebellum and dentate gyrus and serves to translocate neuroblasts, formed in secondary germinal centres, to their definitive adult locations. Radial glia eventually lose their connections with both inner and outer limiting membranes, except for those that persist in the retina as Müller cells, in the cerebellum as Bergmann glia and in the hypothalamus as tanycytes. They can differentiate into neurones as well as astrocytes. They may partially clothe the somata of neighbouring developing neurones (between presumptive synaptic contacts) or similarly enwrap the intersynaptic surfaces of their neurites. Glial processes may expand around intraneural capillaries as perivascular end-feet. Other glioblasts retain an attachment (or form new expansions) to the pia mater, the innermost stratum of the meninges, as pial end-feet. Glioblasts also line the central canal and cavities of the brain as generalized or specialized ependymal cells, but they lose their peripheral attachments. In some situations, such as in the anterior median fissure of the spinal cord, ependymal cells retain their attachments to both the inner and outer limiting membranes. Thus, glia function as perineuronal satellites and provide cellular channels interconnecting extracerebral and intraventricular cerebrospinal fluid, the cerebral vascular bed, the intercellular crevices of the neuropil and the cytoplasm of all neural cell varieties.
Mechanisms of Neural Development
For more than a century the mechanisms that operate during development of the nervous system have been studied experimentally. Although much has been established, answers to many fundamental questions still remain obscure. In recent years, significant advances in our understanding of the development of vertebrates have come from work on amphibian, chicken, mouse and fish embryos and from the production of embryonic chimera (Le Douarin, Teillet and Catala, 1998). A combination of genetic, embryological, biochemical and molecular techniques has been used to elucidate the mechanisms operating in early neural populations.
Histogenesis of the Neural Tube
The wall of the early neural tube consists of an internal ventricular zone (sometimes termed the germinal matrix) abutting the central lumen. It contains the nucleated parts of the pseudostratified columnar neuroepithelial cells and rounded cells undergoing mitosis. The early ventricular zone also contains a population of radial glial cells whose processes pass from the ventricular surface to the pial surface, thus forming the internal and external glia limitans (glial limiting membrane). As development proceeds, the early pseudostratified epithelium proliferates, and an outer layer (the marginal zone), devoid of nuclei but containing the external cytoplasmic processes of cells, is delineated. Subsequently, a middle mantle layer (the intermediate zone) forms as the progeny from the ventricular zone migrate ventriculofugally (see Fig. 3.5).
Most CNS cells are produced in the proliferative zone adjacent to the future ventricular system, and in some regions this area is the only actively mitotic zone. According to the monophyletic theory of neurogenesis, it is assumed to produce all cell types. The early neural epithelium, including the deeply placed ventricular mitotic zone, consists of a homogeneous population of pluripotent cells whose varying appearances reflect different phases in a proliferative cycle. The ventricular zone is considered to be populated by a single basic type of progenitor cell and to exhibit three phases. The cells show an ‘elevator movement’ as they pass through a complete mitotic cycle, progressively approaching and then receding from the internal limiting membrane (Fig. 3.16). DNA replication occurs while the cells are extended and their nuclei approach the pial surface; they then enter a premitotic resting period as the cells shorten and their nuclei pass back toward the ventricular surface. The cells now become rounded close to the internal limiting membrane and undergo mitosis. They then elongate, and their nuclei move toward the outer edge during the postmitotic resting period, after which DNA synthesis commences once more, and the cycle is repeated. The cells so formed may either start another proliferative cycle or migrate outward (i.e. radially) and differentiate into neurones as they approach and enter the adjacent stratum. This differentiation may be initiated as they pass outward during the postmitotic resting period. The proliferative cycle continues with the production of clones of neuroblasts and glioblasts. This sequence of events has been called interkinetic nuclear migration, and it eventually declines. At the last division, two postmitotic daughter cells are produced, and they differentiate at the ventricular surface into ependyma.
The progeny of some of these divisions move away from the ventricular zone to form an intermediate zone of neurones. The early spinal cord and much of the brain stem shows only these three main layers: ventricular, intermediate and marginal zones. However, in the telencephalon, the region of cellular proliferation extends deeper than the ventricular zone, where the escalator movement of interkinetic migration is seen, and a subventricular zone appears between the ventricular and intermediate layers (see Fig. 3.5). Here cells continue to multiply to provide further generations of neurones and glia, which subsequently migrate into the intermediate and marginal zones. In some regions of the nervous system (e.g. the cerebellar cortex) some mitotic subventricular stem cells migrate across the entire neural wall to form a subpial population and establish a new zone of cell division and differentiation. Many cells formed in this site remain subpial in position, but others migrate back toward the ventricle through the developing nervous tissue and finish their migration in various definitive sites where they differentiate into neurones or macroglial cells. In the cerebral hemispheres, a zone termed the cortical plate is formed outside the intermediate zone by radially migrating cells from the ventricular zone. The most recently formed cells migrate to the outermost layers of the cortical plate, so that earlier formed and migrating cells become subjacent to those migrating later. In the forebrain there is an additional transient stratum deep to the early cortical plate, the subplate zone.
Lineage and Growth in the Nervous System
Growth Cones
During development, the growing axons of neuroblasts navigate with precision over considerable distances, often pursuing complex courses to reach their targets. Eventually they make functional contact with their appropriate end-organs (neuromuscular endings, secretomotor terminals, sensory corpuscles or synapses with other neurones). During the outgrowth of axonal processes, the earliest nerve fibres are known to traverse appreciable distances over an apparently virgin landscape, often occupied by loose mesenchyme. A central problem for neurobiologists, therefore, has been understanding the mechanisms of axon guidance (Gordon-Weeks 2000). Axon guidance is thought to involve short-range local guidance cues and long-range diffusible cues, any of which can be either attractive and permissive for growth or repellent and inhibitory. Short-range cues require factors that are displayed on cell surfaces or in the extracellular matrix; for example, axon extension requires a permissive, physical substrate, the molecules of which are actively recognized by the growth cone. They also require negative cues that inhibit the progress of the growth cone. Long-range cues come from gradients of specific factors diffusing from distant targets, which cause neurones to turn their axons toward the source of the attractive signal. The evidence for this has come from in vitro co-culture studies. The floor plate of the developing spinal cord exerts a chemotropic effect on commissural axons that later cross it, whereas there is chemorepulsion of developing motor axons from the floor plate. These forces are thought to act in vivo in concert in a dynamic process to ensure the correct passage of axons to their final destinations and to mediate their correct bundling together en route.
Dendritic Tree
Once growth cones have arrived in their general target area, they have to form terminals and synapses. In recent years, much emphasis has been placed on the idea that patterns of connectivity depend on the death of inappropriate cells. Programmed cell death, or apoptosis, occurs during the period of synaptogenesis if neurones fail to acquire sufficient amounts of specific neurotrophic factors. Coincident firing of neighbouring neurones that have found the appropriate target region might be involved in eliciting the release of these factors, thus reinforcing correct connections. Such mechanisms may explain the numerical correspondence between neurones in a motor pool and the muscle fibres innervated. On a subtler level, pruning of collaterals may give rise to mature neuronal architecture. The projections of pyramidal neurones from the motor and visual cortices, for example, start out with a similar architecture; the mature repertoire of targets is produced by the pruning of collaterals, leading to loss of projections to some targets.
Induction and Patterning of the Brain and Spinal Cord
The regional pattern of the nervous system is induced before and during neural tube closure. Early concepts about regional patterning envisaged that regionalization within mesenchymal populations that transmit inductive signals to the ectoderm imposes a similar mosaic of positional values on the overlying neural plate. For example, transplantation of caudal mesenchyme beneath the neural plate in Amphibia induced spinal cord, whereas rostral mesenchyme induced brain, as assessed by the morphology of the neuroepithelial vesicles. However, later work indicated a more complex scenario in which organizer grafts from early embryos induced mainly head structures, whereas later grafts induced mainly trunk structures. Subsequent molecular data tend to support a model in which neural-inducing factors released by the organizer, such as noggin, chordin and follistatin, neuralize the ectoderm and promote a mainly rostral neural identity. Later secreted signals then act to caudalize this rostral neural tissue, setting up an entire array of axial values along the neural tube. Candidates for these later caudalizing signals include retinoic acid, FGFs, and the WNT secreted proteins, which are present in the paraxial mesenchyme and later in its derivatives, the somites. This combination of signals does not seem to be sufficient to produce the most rostral forebrain structures. Other secreted proteins resident in the rostralmost part of the earliest ingressing axial populations of endoderm and mesenchyme are also capable of inducing markers of forebrain identity from ectodermal cells (Withington, Beddington and Cooke 2001).
Segmentation in the Neural Tube
One mechanism involved in the process of regional differentiation of cell populations within the neural tube is segmentation, which is conspicuous in humans and other vertebrates in the serial arrangement of the vertebrae and axial muscles and in the periodicity of the spinal nerves. In the last century, the possibility that the neural tube might be divided into segments or neuromeres was entertained, but some contended that the bulges observed in the lateral walls of the neural tube were artifacts or were caused by mechanical deformation of the tube by adjacent structures. Recent years have seen a resurgence of interest in this subject and a detailed evaluation of the significance of neuromeres. A series of eight prominent bulges that appear bilaterally in the rhombencephalic wall early in development have been termed rhombomeres (see Fig. 3.3). (Whereas the term neuromere applies generally to putative ‘segments’ of the neural tube, the term rhombomere applies specifically to the rhombencephalon.) Many aspects of the patterning of neuronal populations and the elaboration of their axon tracts conform to a segmental plan, and rhombomeres have now been shown to constitute crucial units of pattern formation. Domains of expression of developmental control genes abut rhombomere boundaries, and perhaps most importantly, single-cell labelling experiments have revealed that cells within rhombomeres form segregated non-mixing populations (Fig. 3.17). The neural crest also shows intrinsic segmentation in the hindbrain and is segregated into streams at its point of origin in the dorsal neural tube. This may represent a mechanism whereby morphogenetic specification of the premigratory neural crest cells is conveyed to the pharyngeal arches. Although these segmental units lose their morphological prominence with subsequent development, they represent the fundamental ground plan of this part of the neuraxis, creating a series of semiautonomous units within which local variations in patterning can develop. The consequences of early segmentation for later developmental events, such as the formation of definitive neuronal nuclei within the brain stem, and of peripheral axonal projections remain to be explored.
(Modified by permission from Annual Review of Cell and Developmental Biology, Vol. 8. 1992. www.annualreviews.org.)
Genes such as the Hox and Pax gene families, which encode transcription factor proteins, show intriguing expression patterns within the nervous system. Genes of the Hox-b cluster, for example, are expressed throughout the caudal neural tube and up to discrete limits in the hindbrain that coincide with rhombomere boundaries. The ordering of these genes within a cluster on the chromosome (5′–3′) is the same as the caudal-to-rostral limits of expression of consecutive genes. This characteristic pattern is surprisingly similar in fish, frogs, birds and mammals. Hox genes play a role in patterning of not only the neural tube but also much of the head region, consistent with their expression in neural crest cells and within the pharyngeal arches. Disruption of the Hox a-3 gene in mice mimics DiGeorge syndrome, a congenital human disorder characterized by the absence (or near absence) of the thymus, parathyroid and thyroid glands; hypotrophy of the walls of the arteries derived from the aortic arches and subsequent conotruncal cardiac malformations. Some Pax genes are expressed in different dorsoventral domains within the neural tube. Pax-3 is expressed in the alar lamina, including the neural crest, whereas Pax-6 is expressed in the intermediate plate. The Pax-3 gene has the same chromosomal localization as the mouse mutation Splotch and the affected locus in the human Waardenburg’s syndrome, both of which are characterized by neural crest disturbances with pigmentation disorders and occasional neural tube defects. Both Hox and Pax genes have restricted expression patterns with respect to the rostrocaudal and dorsoventral axes of the neural tube, consistent with roles in positional specification. (For reviews of the expression patterns of these genes, see Krumlauf et al 1993.)
Whereas craniocaudal positional values are probably conferred on the neuroepithelium at the neural plate or early neural tube stage, dorsoventral positional values may become fixed later. The development of the dorsoventral axis is heavily influenced by the presence of the underlying notochord. The notochord induces the ventral midline of the neural tube, the floor plate. This specialized region consists of a strip of non-neural cells with distinctive adhesive and functional properties. Notochord and floor plate together participate in inducing the differentiation of the motor columns. Motor neurone differentiation occurs early, giving some support to the idea of a ventral-to-dorsal wave of differentiation. The notochord–floor plate complex may also be responsible for allotting the values of more dorsal cell types within the tube (see Fig. 3.7). For example, the dorsal domain of Pax-3 expression extends more ventrally in embryos experimentally deprived of notochord and floor plate, whereas grafting an extra notochord adjacent to the dorsal neural tube leads to the repression of Pax-3 expression.
Peripheral Nervous System
Somatic Nerves
Spinal Nerves
Each spinal nerve is connected to the spinal cord by a ventral root and a dorsal root (Fig. 3.18). The fibres of the ventral roots grow out from cell bodies in the anterior and lateral parts of the intermediate zone. These pass through the overlying marginal zone and external limiting membrane. Some enter the myotomes of the somites, and some penetrate the somites, reaching the adjacent somatopleure; in both sites they ultimately form the α-, β- and γ-efferents. At appropriate levels these are accompanied by the outgrowing axons of preganglionic sympathetic neuroblasts (segments T1–L2) or preganglionic parasympathetic neuroblasts (S2–4).
The fibres of the dorsal roots extend from cell somata in dorsal root ganglia into the spinal cord and also into the periphery. Neural crest cells are produced continuously along the length of the spinal cord, but gangliogenic cells migrate only into the rostral part of each somitic sclerotome, where they condense and proliferate to form a bilateral series of oval-shaped primordial spinal ganglia (dorsal root ganglia; see Fig. 3.12). Negative factors in the caudal sclerotome deter neural crest from entering. The rostral sclerotome has a mitogenic effect on the crest cells that settle within it. From the ventral region of each ganglion, a small part separates to form sympathochromaffin cells, whereas the remainder becomes a definitive spinal ganglion (dorsal root ganglion). The spinal ganglia are arranged symmetrically at the sides of the neural tube and, except in the caudal region, are equal in number to the somites. The cells of the ganglia, like the cells of the intermediate zone of the early neural tube, are glial and neuronal precursors. The glial precursors develop into satellite cells (which become closely applied to the ganglionic nerve cell somata), Schwann cells and possibly other cells. The neuroblasts, which are initially round or oval, soon become fusiform, and their extremities gradually elongate into central and peripheral processes. The central processes grow into the neural tube as the fibres of dorsal nerve roots, and the peripheral processes grow ventrolaterally to mingle with the fibres of the ventral root, thus forming a mixed spinal nerve. As development proceeds, the original bipolar form of the cells in the spinal ganglia changes, and the two processes become approximated until they ultimately arise from a single stem to form a unipolar cell. The bipolar form is retained in the ganglion of the vestibulocochlear nerve.
Cranial Nerves
Cranial nerves may contain motor, sensory or both types of fibres. With the exception of the olfactory and optic nerves, the cranial nerves develop in a manner similar in some respects to components of the spinal nerves. The somata of motor neuroblasts originate within the neuroepithelium; those of sensory neuroblasts are derived from the neural crest, with additional contributions in the head from ectodermal placodes (Fig. 3.19).
The motor fibres of the cranial nerves that project to striated muscle are the axons of cells originating in the basal plate of the midbrain and hindbrain. The functional and morphological distinction between the neurones within these various nerves is based on the types of muscle innervated. In the trunk, the motor roots of the spinal nerves all emerge from the spinal cord close to the ventral midline, to supply the muscles derived from the somites. In the head, the motor outflow is traditionally divided into two pathways (see Figs. 3.2B, 3.19). General somatic efferent neurones exit ventrally in a similar manner to those of the spinal cord. Thus the oculomotor, trochlear, abducens and hypoglossal nerves parallel the organization of the somatic motor neurones in the spinal cord. The second motor component, the special branchial efferent, consists of the motor parts of the trigeminal, facial, glossopharyngeal and vagus nerves, which supply the pharyngeal (branchial) arches and the accessory nerve. All these nerves have nerve exit points more dorsally placed than in the somatic motor system.
The cranial sensory ganglia are derived in part from the neural crest and in part from cells of the ectodermal placodes (see Figs. 3.13, 3.19). Generally, neurones distal to the brain are derived from placodes, and proximal ones are derived from the neural crest (see Fig. 3.19). Supporting cells of all sensory ganglia arise from the neural crest. The most rostral sensory ganglion, the trigeminal, contains both neural crest– and placode-derived neurones that mediate general somatic afferent functions. The same applies to the more caudal cranial nerves (facial, glossopharyngeal, vagus), but the two cell populations form separate ganglia in the case of each nerve. The proximal series of ganglia is derived from neural crest (forming the proximal ganglion of the facial nerve, the superior ganglion of the glossopharyngeal nerve and the jugular ganglion of the vagus nerve); the distal series is derived from placodal cells (forming the geniculate ganglion of the facial nerve, the petrosal ganglion of the glossopharyngeal nerve and the nodose ganglion of the vagus nerve). These ganglia contain neurones that mediate special, general visceral and somatic afferent functions. The vestibulocochlear nerve has a vestibular ganglion that contains both crest and placodal cells and an acoustic ganglion from placodal neurones only; it conveys special somatic afferents.
Autonomic Nervous System
In the trunk at neurulation, neural crest cells migrate from the neural epithelium to lie transitorily on the fused neural tube. Thereafter, crest cells migrate laterally and then ventrally to their respective destinations (see Fig. 3.12). In the head, the neural crest cells migrate prior to neural fusion, producing a vast mesenchymal population as well as autonomic neurones.
Neurones of the enteric nervous system are described as arising from the vagal crest—that is, the neural crest derived from somite levels 1 to 7, and the sacral crest caudal to the twenty-eighth somite. At all these levels the crest cells also differentiate into glial-like support cells alongside the neurones (Fig. 3.20).
Sympathetic Ganglia
Neural crest cells migrate ventrally within the body segments to penetrate the underlying somites and continue to the region of the future paravertebral and prevertebral plexuses, notably forming the sympathetic chain of ganglia as well as the major ganglia around the ventral visceral branches of the abdominal aorta (see Figs. 3.12, 3.20).
The local environment is the major factor that controls the appropriate differentiation of the presumptive autonomic ganglion neurones. The factors responsible for subsequent adrenergic, cholinergic or peptidergic phenotype have yet to be identified, although it has been proposed that fibronectin and basal lamina components initiate adrenergic phenotypical expression at the expense of melanocyte numbers. Cholinergic characteristics are acquired relatively early, and the appropriate phenotypical expression may be promoted by cholinergic differentiation factor and CNTF.
Enteric Nervous System
The enteric nervous system is derived from the neural crest. The axial levels of crest origin are shown in Figure 3.20. Premigratory neural crest cells are not prepatterned for specific axial levels; rather, they attain their axial value as they leave the neuraxis. Once within the gut wall, there is a regionally specific pattern of enteric ganglia formation that may be controlled by the local splanchnopleuric mesenchyme. Cranial neural crest from somite levels 1 to 7 contributes to the enteric nervous system, forming both neuroblasts and glial support cells.