Section I / General
Developmental
Fig. 3.9  Anencephaly.
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 hypophysis cerebri consists of the adenohypophysis and the neurohy-
pophysis. Prior to neurulation the cell populations that give rise to these two
portions of the pituitary gland are found next to each other within the rostral
portion of the floor of the neural plate and the contiguous midline neural
fold. As neurulation proceeds the future neurohypophysis remains within the
floor of the prosencephalon, and the cells of the future adenohypophysis are
displaced into the surface ectoderm, where they form the hypophysial
placode.
The most rostral portion of the neural plate, which will form the hypo-
thalamus, 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 noto-
chord 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 pos-
terior 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
38
A Fate map B potentials
PROSENCEPHALON
MESENCEPHALON
ROSTRAL
RHOMBENCEPHALON
S1
CAUDAL
S5
S7
CERVICAL
SPINAL CORD
S18
THORACIC
S24 SPINAL CORD
S28
LUMBOSACRALL
SPINAL CORD
Mesectoderm Sensory ganglion
Ectomesenchyme Sympathetic ganglia
parasympathetic
ganglia
Fig. 3.10  A, Fate map along the neural crest of the presumptive territories that
yield the ectomesenchyme; the sensory, parasympathetic and sympathetic
ganglia; and neural crest–derived mesenchyme in normal development.
B, Developmental potentials for the same cell types. If neural crest cells from
any level of the neural axis are implanted in the appropriate sites of a host
embryo, they can give rise to almost all the cell types forming the various
kinds of peripheral nervous system ganglia. This is not true for the neural
crest–derived mesenchyme, whose precursors are confined to the cephalic
area of the crest down to the level of somite (S) 5.
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.
A small endodermal diverticulum, called Sessel’s pouch, projects toward
the brain from the cranial end of the foregut, immediately caudal to the buc-
copharyngeal membrane. In some marsupials this pouch forms a part of the
hypophysis, but in humans it apparently disappears entirely.
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 pro-
cesses 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 neu-
rohypophysis as the pars intermedia.
 Chapter 3 / Development of the Nervous System

A Hypothalamus B
Adenohypophysis
Nasal cavity

Chapter 3
Olfactory
placode

PROSENCEPHALON
Frontonasal ectoderm
Telencephalon

Eye

Neurohypophysis
Nasal cavity
Maxilla
Optic placode
Philtrum

Thalamus Primary
Epiphysis palate

Secondary
palate

MESENCEPHALON
Maxillo-mandibulary

trigeminal placode.
Mandible

ectoderm and
Mesencephalon
Trigeminal
Derived from tissues
placode
of several branchial
arches not included
in this diagram

Fig. 3.11  Fate map of the rostral region of the neural primordium as established by the quail–chick chimera system. A, The various territories yielding a rostral head are
indicated on the neural plate and neural fold of a one- to three-somite embryo. B, Results obtained in the avian embryo have been extrapolated to the human head.
Thus, the neural fold area coloured green yields the epithelium of the rostral roof of the mouth, the nasal cavities and part of the frontal area.

Dorsal root NEURAL CREST CELLS PLACODES

Ganglia and glia Neural crest cells
of the sympathetic ganglia and glia
trunk Olfactory
Optic
Melanocytes Location Location
derivatives derivatives

Ciliary
Trigeminal
Trigeminal

Distal VII
Proximal VII (root) (geniculate)
Ethmoidal
Sphenopalatine Distal IX (petrosal)
Proximal IX
(superior) Vestibulo-acoustic (otic)
Proximal X Distal X (nodose)
(jugular)

Preaortic Enteric Medulla of

ganglia and ganglia and
suprarenal
glia glia gland
Fig. 3.12  Migration routes taken by neural crest cells in the trunk.
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
Glial cells that support neurones in the CNS and PNS are derived from three
lineages: the neuroectoderm of the neural tube, the neural crest and the
angioblastic mesenchyme. In the CNS, cells of the proliferating ventricular
zone give rise to astrocyte and oligodendrocyte cell lines. After the prolifera-
tive phase, the cells remaining at the ventricular surface differentiate into
ependymal cells, which are specialized in many regions of the ventricular
Fig. 3.13  Positions of the neural crest and placodal cells in a stage 9.5 chick
embryo. Neural crest cells are shown in the midline in green. Placodes are
more laterally placed in grey. The otic placode is dorsolateral to the
rhombencephalon, the trigeminal placode is placed intermediately and the
epibranchial placodes (for facial, glossopharyngeal and vagus cranial nerves) are
placed ventrolaterally and dorsal to the future pharyngeal grooves. (From
D’Amico-Martel, A., Noden, D.M. 1983. Am. J. Anat. 166, 445–468. Reprinted by
permission of Wiley-Liss Inc.)
system as circumventricular organs. In the PNS, neural crest cells produce
Schwann cells and astrocyte-like support cells in the enteric nervous system.
Angioblastic mesenchyme gives rise to a variety of blood cell types, including
circulating monocytes that infiltrate the brain as microglial cells later in
development.
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

39
Section I / General

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 perineu-
ronal satellites and provide cellular channels interconnecting extracerebral
and intraventricular cerebrospinal fluid, the cerebral vascular bed, the inter-
cellular crevices of the neuropil and the cytoplasm of all neural cell
varieties.
Microglia appear in the CNS after it has been penetrated by blood vessels
and invade it in large numbers from certain restricted regions. From there they
spread in what have picturesquely been called ‘fountains of microglia’ to
extend deeply among the nervous elements.

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.
Fig. 3.14  Scanning electron micrograph of the roof of the pharynx showing the In recent years, significant advances in our understanding of the development
invagination of placodal ectoderm to form the adenohypophysis (Rathke’s of vertebrates have come from work on amphibian, chicken, mouse and fish
pouch). (Photograph by P. Collins; printed by S. Cox, Electron Microscopy Unit, embryos and from the production of embryonic chimera (Le Douarin, Teillet
Southampton General Hospital.) and Catala, 1998). A combination of genetic, embryological, biochemical and
molecular techniques has been used to elucidate the mechanisms operating
in early neural populations.
Hypophyseal recess Midbrain
A The CNS has a fundamental structure of layers and cells that are all derived
(Rathke’s pouch)
from a pluripotential neuroepithelium. Developing neuroblasts produce axons
Forebrain that traverse great distances to reach their target organs. Within the CNS they
form myriad connections with other neuroblasts in response to locally
secreted neurotrophins. The brain and spinal cord reveal an intrinsic metamer-
ism, induced rostrally by genes and caudally by inductive influences from
adjacent structures.

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 con-
Notochord tains the nucleated parts of the pseudostratified columnar neuroepithelial
Foregut cells and rounded cells undergoing mitosis. The early ventricular zone also
Pericardium
contains a population of radial glial cells whose processes pass from the
Oropharyngeal ventricular surface to the pial surface, thus forming the internal and external
membrane glia limitans (glial limiting membrane). As development proceeds, the early
pseudostratified epithelium proliferates, and an outer layer (the marginal
B zone), devoid of nuclei but containing the external cytoplasmic processes of
Diencephalon Midbrain Posterior cells, is delineated. Subsequently, a middle mantle layer (the intermediate
lobe
Interventricular zone) forms as the progeny from the ventricular zone migrate ventriculofu-
foramen gally (see Fig. 3.5).
Hypophysis
Anterior Most CNS cells are produced in the proliferative zone adjacent to the
lobe
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
Notochord deeply placed ventricular mitotic zone, consists of a homogeneous popula-
tion 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
Telencephalon show an ‘elevator movement’ as they pass through a complete mitotic cycle,
Degenerated progressively approaching and then receding from the internal limiting mem-
oropharyngeal
brane (Fig. 3.16). DNA replication occurs while the cells are extended and their
membrane
First arch nuclei approach the pial surface; they then enter a premitotic resting period
ventral (mandibular) part as the cells shorten and their nuclei pass back toward the ventricular surface.
Fig. 3.15  A and B, Sagittal sections of heads of early embryos showing the first The cells now become rounded close to the internal limiting membrane and
stages in the development of the hypophysis. undergo mitosis. They then elongate, and their nuclei move toward the outer
edge during the postmitotic resting period, after which DNA synthesis com-
mences once more, and the cycle is repeated. The cells so formed may either
mater, and inward, to form the inner limiting membrane around the central start another proliferative cycle or migrate outward (i.e. radially) and differ-
cavity. The geometry of these cells may provide contact guidance paths for entiate into neurones as they approach and enter the adjacent stratum. This
cell migrations, both neuroblastic and glioblastic. A secondary radial glial differentiation may be initiated as they pass outward during the postmitotic
scaffold is formed in the late-developing cerebellum and dentate gyrus and resting period. The proliferative cycle continues with the production of
serves to translocate neuroblasts, formed in secondary germinal centres, to clones of neuroblasts and glioblasts. This sequence of events has been called

40

Intermediate zone
DNA synthesis
Ventricular in this part of
zone/ ventricular
germinal zone
matrix
Ventricular surface
Phase of
cell cycle
G2 phase
G1 phase
S phase
S phase
Mitosis
Fig. 3.16  Cell cycle in the ventricular zone of the developing neural tube. The
nuclei of the proliferating stem cells show interkinetic migration. (From Fujita, S.
1963. J. Comp. Neurol. 120, 37–42. Reprinted by permission from Wiley-Liss Inc.)
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, inter-
mediate 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 differentia-
tion. 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
Neurones come from two major embryonic sources: CNS neurones originate
from the pluripotential neural plate and tube, whereas ganglionic neurones
originate from the neural crest and ectodermal placodes. The neural plate
also provides ependymal and macroglial cells. Peripheral Schwann cells and
chromaffin cells arise from the neural crest. The origins and lineages of cells
in the nervous system have been determined experimentally by the use of
autoradiography, by microinjection or retroviral labelling of progenitor cells
and in cell culture.
During development, neurones are formed before glial cells. The timing of
events differs in various parts of the CNS and between species. Most neurones
are formed prenatally in mammals, but some postnatal neurogenesis does
occur (e.g. the small granular cells of the cerebellum, olfactory bulb and hip-
pocampus, and neurones of the cerebral cortex). Gliogenesis continues after
birth in periventricular and other sites. Autoradiographic studies have shown
that different classes of neurones develop at specific times. Large neurones,
such as principal projection neurones, tend to differentiate before small ones,
such as local circuit neurones. However, their subsequent migration appears
to be independent of the time of their initial formation. Neurones can migrate
extensively through populations of maturing, relatively static cells to reach
their destination; for example, cerebellar granule cells pass through a layer of
Purkinje cells en route from the external pial layer to their final central posi-
tion. Later, the final form of their projections, their cell volume and even their
continuing survival depend on the establishment of patterns of functional
connection.
Chapter 3 / Development of the Nervous System
Initially, immature neurones, termed neuroblasts, are rotund or fusiform.
Their cytoplasm contains a prominent Golgi apparatus, many lysosomes, gly-
cogen and numerous unattached ribosomes. As maturation proceeds, cells
send out fine cytoplasmic processes that contain neurofilaments, microtu-
bules and other structures, often including centrioles at their bases where
microtubules form. Internally, endoplasmic reticulum cisternae appear, and
Chapter 3
attached ribosomes and mitochondria proliferate, whereas the glycogen
content progressively diminishes. One process becomes the axon, and other
processes establish a dendritic tree. Axonal growth, studied in tissue culture,
may be as much as 1 mm per day.
Growth Cones
Ramón y Cajal (1890) was the first to recognize that the expanded end of an
axon, the growth cone, is the principal sensory organ of the neurone. The
growing tips of neuroblasts have been studied extensively in tissue culture.
Classically, the growth cone is described as an expanded region that is con-
stantly active, changing shape, extending and withdrawing small filopodia and
lamellipodia that apparently ‘explore’ the local environment for a suitable
surface along which extension can occur. These processes are stabilized in
one direction, determining the direction of future growth, and after consoli-
dation of the growth cone, the exploratory behaviour recommences. This
continuous cycle resembles the behaviour at the leading edge of migratory
cells such as fibroblasts and neutrophils. The molecular basis of this behaviour
is the transmission of signals external to the growth cone via cell surface
receptors to the scaffolding of microtubules and neurofilaments within the
axon. Growing neuroblasts have a cortex rich in actin associated with the
plasma membrane, along with a core of centrally located microtubules and
sometimes neurofilaments. The assembly of these components, as well as the
synthesis of new membrane, occurs in segments distal to the cell body and
behind the growth cone, although some assembly of microtubules may take
place near the cell body.
The driving force of growth cone extension is uncertain. One possible
mechanism is that tension applied to objects by the leading edge of the
growth cone is mediated by actin, and local accumulations of F-actin redirect
the extension of microtubules. Under some culture conditions, growth cones
can develop mechanical tension, pulling against other axons or the substratum
to which they are attached. It is possible that tension in the growth cone acts
as a messenger to mediate the assembly of cytoskeletal components.
Adhesion to the substratum appears to be important for consolidation of the
growth cone and elaboration of the cytoskeleton in that direction.
During development, the growing axons of neuroblasts navigate with preci-
sion 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 cor-
puscles 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 mes-
enchyme. A central problem for neurobiologists, therefore, has been under-
standing 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 neu-
rones 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 inappropri-
ate cells. Programmed cell death, or apoptosis, occurs during the period of
synaptogenesis if neurones fail to acquire sufficient amounts of specific neu-
rotrophic 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
41