DEVBIOL

LECTURE 6: NEURULATION
Gliceria B. Ramos| 1st TERM | A.Y. 2022-2023

OUTLINE n. Crest cells and detach from the epidermal

I. A IV. B
cells
II.
○ Expression of N-cadherin and E-cadherin
adhesion proteins during neurulation in
Xenopus.
○ In normal development of the neural plate
stage:
■ N-cadherin is seen in the neural
plate
NEURULATION OVERVIEW ■ E-cadherin is seen on the
● Laying down the rudiments of the CNS (brain and presumptive epidermis.
spinal cord) ○ Eventually, the N-cadherin-bearing neural
● Includes the formation of the neural crest cells cells separate from the
(NCCs) E-cadherin-containing epidermal cells.
○ Mesenchymal type
○ Can migrate to the different parts of the body
and give rise to different structures
○ Form the ganglia
● Defining the various brain divisions
● Starting organogenesis
● Continuing the establishment of the embryonic axis
○ Started at gastrula stage

RECAP OF GASTRULATION FORMATION OF THE NEURAL GROOVE AND

● Formation of the chordamesoderm in amphibians NEURAL FOLDS
● Formation of the axial mesoderm (notochordal ● Neural plate deepens and forms the neural groove
process in mammals) for chicks ● Its margins are increased by
○ Have an overlying ectoderm from epiblast ○ Convergent extension (cell rows shift into
● Generation of the chordamesoderm/ axial mesoderm/ one another and form the neural folds)
notochordal process ● Convergence
○ Have inductive effect ○ Coupled with convergent elongation and
○ Facilitate neural induction convergent thickening
○ Folds moving to the midline
NEURULATION ○ Formation of a v-shaped cavity - neural
● Neural induction groove
○ The specification of the dorsal ectoderm by
the inducing effect of the chordamesoderm/
axial mesoderm/ notochordal process
○ Dorsal ectoderm → neural plate
● Formation of the neural plate
○ Morphogenic changes: shaping, bending,
and folding neural plate bends and edges
elevate
○ Epithelial cell behavior: bending, folding
○ Cell behavior: cell shaping

*blue epidermal cells

*green neural crest cells

FORMATION OF THE TUBE BY CLOSURE OF THE

NEUROPORE
● The neural folds then merge, cut off the neural
groove, and form the neural tube
● The closure of the neural tube disconnects the neural
crest cells from the epidermis
● Migration of the neural crest cells is the final step in
the neural tube closure
○ The neural crest cells have neither cadherin,
and they disperse
■ Cell adhesion proteins
■ Can easily disperse and migrate
*Neural plate - Tall columnar cells distinguish themselves from
surrounding neural crest cells and presumptive epidermal cells
● 3 Cell Populations:
○ Presumptive neural plate cells
■ Undergo columnarization (low
columnar cell to tall columnar cells)
○ Presumptive neural crest cells
○ Presumptive epidermal cells
■ Flat squamous cells
● Thickening is the first phase of neurulation
● Neural plate formation
○ P. Neural plate cells express for the genes ● Other mesoderm cells differentiate into dorsal
encoding for N cadherin, neural cell mesoderm -> somites
adhesion protein ○ Dorsal mesoderm becomes paraxial
○ P. epidermal cells express the gene mesoderm and the ???
encoding for E-cadherin, epithelial cell ○ Other mesoderm will thicken to form
adhesion protein notochordal process or axial mesoderm
○ Eventually, these CAMs (N-cadherin and
E-cadherin) will clump the p. Epidermal and

1

● Meeting of neural folds completely at the midline
○ Epidermal cells meet in the midline
○ Neural crest cells detach underneath
epidermal cells and over the roof of neural
groove
DORSAL-VENTRAL SPECIFICATION OF THE
NEURAL TUBE
● Two primary signaling centers
○ Ectodermal cells of the epidermis produce
BMP4 and BMP7 (TGF-B) -> roof of the
neural tube is exposed (A)
○ Notochordal cells produce Sonic hedgehog
protein -> floor of the neural tube is exposed
(B)
■ Floor plate becomes a secondary
signaling center in response to
sonic hedgehog
● Secondary signaling centers established within the
neural tube (C)
○ Roof plate cells express/secrete BMP4
○ Floor plate cells express/secrete Sonic
hedgehog
*signaling centers will determine formation of dorsal and
ventral side depending on the concentration gradients
● Two concentration gradients are established (D);
gradients of TGF-B and Shh
○ Different concentration gradients activate the
expression of different sets of genes
○ -> Different exposure level of cells result to
different identities
○ -> cells then differentiate to become
interneurons and motorneurons
■ Motor- more ventral
■ Sensory- more dorsal
■ Interneurons- in between gradients
● Dorsoventral axis of embryo is already established
during neural tube formation
● Notochord eventually degenerates
○ Persists as nucleus pulposus of the
vertebral discs
TEMPORARY OPENING OF NEURAL TUBE
● The neural tube is temporarily open at both ends
(anterior and posterior)
● This allows the passage of amniotic fluid
● Anterior neuropore
○ Opening at head end
○ Region of brain development
● Posterior neuropore
○ Opening at caudal end
○ Region of spinal cord development
● Neurocoele will form the ventricles of CNS
○ Lateral ventricles, 3rd ventricle, and 4th
ventricle
● Anterior neuropore closes at 24-26 day of
development
● Posterior neuropore closes at 28-day development
Defects in the Neural Tube
● Failure of closure in anterior or posterior neuropore
causing embryonic conditions:
● Anencephaly - failure of Anterior neuropore to close
○ Missing forebrain portions; fatal
○ Compressed brain or abnormally small brain
● Spina Bifida - failure of posterior neuropore to close
○ Spina Bifida occulta - mild; no pain; no
neurological disorder
■ Presents as a dimple dorsally
○ Spina Bifida cystica - severe; neurological
disorders
■ Bulging of spinal cord
PRIMARY VS. SECONDARY NEURULATION
● Primary - development of the neural tube induced by
the notochord and the mesoderm
○ In human embryo: ends at 4th week of
embryonic development with the closure of
the posterior neuropore
● Secondary - caudal end of neural tube develops into
the neural notochord, then canalized
○ Starts off as a solid rod called the medullary
cord, then cavitates
○ This undergo epithelialization
○ First half and second half that is
epithelialized then face each other in a
juxtapose position
○ The space inbetween will form cavity to
become the neural tube
○ In avians, secondary neurulation starts at the
level of the 25-somite
○ In humans, occurs at the level of the
35-somite, or 6th week of embryonic
development
EXTENT OF USE OF PRIMARY OR SECONDARY
NEURULATION VARY ON VERTEBRATE CLASS
● Fishes: exclusively on secondary neurulation
○ Formation of keel
● Birds: both
○ anterior region - Primary neurulation
○ Posterior region to 25th-27th somite -
Secondary neurulation
● Amphibians: mostly primary neurulation
○ Tail end - secondary neurulation
● Humans: Secondary neurulation around 35th
somite
2

Early human brain development
AVIAN NEURULATION
● Primary neurulation: neural tube formation in the chick
embryo
○ (A, 1) Cells of the neural plate can be
distinguished as elongated cells in the dorsal
region of the ectoderm.
■ Folding begins as the medial
neural hinge point (MHP) cells
anchor to notochord and change
their shape, while the presumptive
epidermal cells move towards the
center.
○ (B, 2) The neural folds are elevated as
presumptive epidermis continues to move
toward the dorsal midline.
○ (C, 3) Convergence of the neural folds
occurs as the dorsolateral hinge point
(DHLP) cells become wedge-shaped and
epidermal cells push toward the center
○ (D, 4) The neural folds are brough into
contact with one another, and the neural
crest cells link the neural tube with the
epidermis.
■ The neural crest cells then
disperse, leaving the neural tube
separate from the epidermis.
Neural tube formation does not occur simultaneously
throughout the ectoderm
● Starts out as elongated primitive streak
● Neurulation on the cephalic end → well-advanced
● Neurulation in the caudal end → still undergoing (late
gastrulation)
● Regionalization of the tube
DIFFERENTIATION OF THE NEURAL TUBE
● Occurs simultaneously in 3 ways:
● Gross anatomical level
○ NT and neurocoel → chambers of the brain
and spinal cord
● Tissue level
○ Cells in the wall of NT → functional regions
of the brain and spinal cord
● Cellular level
○ Neuroepithelial cells → numerous types of
nerve cells (neurons) & supportive cells
(neuroglial cells)
● The three primary brain vesicles are subdivided as
development continues
Adult derivatives
● Forebrain (Prosencephalon)
○ Telencephalon
○ Diencephalon
● Midbrain (Mesencephalon)
○ Mesencephalon
● Hindbrain (Rhombencephalon)
○ Metencephalon
○ Myelencephalon
○ In the hindbrain, distinct subdivisions or
swellings are formed called rhombomeres
■ Rhombomeres - periodic swellings
on the rhombencephalon; with the
neural crest cells and will determine
where cranial nerves will arise
■ Will form the ganglia → associated
neural crest cells, over the roof of
the rhombencephalon and
underneath the epidermal cell, will
determine the position and location
of the cranial nerves
■ R2 = CN5
■ R4 = CN7 and CN8
■ R6 = CN9
■ R3 = will undergo apoptosis
■ R5 = will undergo apoptosis
○ A ganglion is an aggregate of nerve cell
bodies (collection of axonal processes) and
occur as thickening of neural crest cells
3
DEVELOPMENTAL EVENTS OF THE HUMAN EMBRYO

F. 23-day old embryo (approximately)

○ Pericardial bulge in each side of the midline
in the cephalic part
○ Anterior neuropore and posterior neuropore
allow passage of amniotic fluid

A. 16-day old embryo

○ Visible primitive streak with the primitive
node
B. 18-day old embryo
○ A groove at the midline is visible
○ Pear shaped; cephalic end broader than
caudal end

G. 25-day old embryo (approximately)

○ 18-20 somites
○ 14-somite stage; bulging pericardial
structure;
○ 1st and 2nd pharyngeal arches forming
○ Closure of cranial neuropore
C. 19-day old embryo (approximately)
○ Rewind from Gastrulation:
■ In the gastrula stage, the epiblast
splits into two — amniotic ectoderm
and epiblastic embryonic ectoderm
■ Amniotic ectoderm
● It forms a helmet around
the epiblastic ectoderm of
the developing embryo
● It encloses the amniotic
cavity that has the fluid
○ Amnion is cut to expose dorsal view; H. 28-day old embryo (approximately)
■ Neural plate visible ○ 25 somites
● Thickening ○ 3 pharyngeal arches formed
● Undergoing epithelial ○ Sensory placodes formed (LENS & OTIC)
columnarization ○ Closure of the posterior neuropore

NEURAL INDUCTION
As a multistep process

D. 20-day old embryo (approximately)

○ Neural groove & fold formed;
○ Somites forming lateral to the neural
structures
■ 3 distinct pairs of somites are
formed
○ Fusion of the neural fold begins in the
cervical region (level of 5th somite)
E. 22-day old embryo (approximately)
○ 7 distinct somites on the sides of the closed
neural tube
○ 5th somite
■ Starts the closure of neural fold
● Started as early as cytoplasmic movements in the
fertilized egg/oocyte
○ With fertilization, there is cortical movement,
granules moving in the point of sperm entry
— dragging the pigment granules which
creates the gray crescent region in the
dorsal part.
○ The gray crescent region determines the
dorsal side of the embryo
○
● In later cleavage, only dorsal blastomeres
(micromeres) are able to induce animal cells to form
dorsal mesoderm
○ Based on research reports, early on, there is
already a bias between the ventral cells
(macromeres) than the animal cells
■ The ventral cells are expressing
more of the molecular marker for
epidermal cells — Epi1 marler

4
 ■ Animal cells are also expressing
Epi1, BUT only at a lower level and
eventually to inhibited expression
* In the inductive process, there must be a synthesis of
competence factor to respond to the incoming signals.
● Remember that at neurulation, there is establishment
of the body axis; dorsal to the lateral, dorsal to medial,
and dorsal to ventral

● Dorsal mesoderm will form the dorsal lip of the

blastopore (organizer)
○ Sends planar induction signals at gastrula
stage
○ With further involution, it will form the
chordamesoderm
○ At DLB, the chief cell organizer of the
embryo is the reason of chordamesoderm’s
formation
● Chordamesoderm sends vertical signals to the
overlying prospective neuroectoderm/dorsal
ectoderm From the point of the primitive node and upwards, are the
neural structures or embryonic structures
1. Neuroectoderm is primed to respond to signal stimuli
2. Inductive signals (from chordamesoderm) may act by Three types of mesoderm are the:
disinhibition (interfere with inhibitory signal that 1. paraxial mesoderm at the side of the neural tube or to the
normally blocks a default program) side of the axial mesoderm and the neural tube
AXIS INDUCTION
Axis induction “Para”=side
o vertebrate embryonic cells, after midblastula
transition, develop dorsal organ rudiments unless 2. Intermediate mesoderm
they are told differently 3. lateral plate mesoderm

● Bone morphogenetic protein 4 (BMP 4) as the natural * after this is the establishment of the body axes
inhibitor of dorsal axis formation * Body axes formation

The inducer or natural inhibitor is BMP4 in axis induction

Dorsal lip of blastopore expresses the goosecoid gene, which

codes for a transcription factor that regulates the activity of
chordin and noggin; if chordin and noggin is activated, then it
inactivates BMP4, so when the BMP4 is inactive, dorsal
development is allowed which will lead to the establishment of
the dorsal mesoderm but when BMP4 is active, ventral
development is favored, and there will be formation of ventral
mesoderm

Neurulation or the establishment of the dorsal side is by

disinhibition or inhibition of BMP4
GOOSECOID GENE AT THE ORGANIZER

● Goosecoid codes for a transcription factor so it can

regulate the activity of chordin and noggin at the
organizer, and they can inactivate BMP4, and when
BMP4 is inactive, it allows dorsal development, and
neurulation can proceed, thereby, forming the neural
ectoderm and dorsal ectoderm ; some of the dorsal
mesoderm will move sideways to form the paraxial
mesoderm while others remain at the midline to form
the axial mesoderm, and the notochordal process, so
tue dorsal side is established

● Whereas, in the case that BMP4 is active, then it

favors ventral development, which will lead to the
ventral epidermis differentiating into the intermediate
mesoderm and the lateral plate mesoderm