Lecture 18: Chapter 23 - (Development I)

A. Anatomy of the Developing Embryo

Three tissues make up the embryo: ectoderm, mesoderm, and endoderm.

Ectoderm gives rise to the nervous system and skin.

Zygote  blastula  blastocoel  neurogenic region  neural plate

- Neurulation: The neural plate forms the neural groove. The mesoderm forms the notochord, which creates the
neural tube from the groove.
- Some of the neural ectoderm is pinched off and becomes the Neural Crest
- The neural tube becomes the CNS
- The neural crest becomes the PNS

The Neural Tube forms 3 swellings called the primary vesicles:

1. Prosencephalon: forebrain
2. Mesencephalon: midbrain
3. Rhombencephalon: hindbrain

Forebrain: Secondary vesicles sprout off on both sides (optic and telencephalic vesicles)
Diencephalon: the structure between the 2 vesicles

B. Six Stages of Brain Development

1. Neurogenesis:

- Both neurons and glial cells proliferate in the Neural tube around the central canal
- The central canal eventually becomes the vesicles
- Vesicle walls have 2 layers: the marginal and ventricular zones
- The Ventricular Zone: lines the inside of each vesicle
- The Marginal Zone: faces the overlying pia

Cell Proliferation:
1st Position: A cell in the ventricular zone extends a process that reaches upward toward the pia
2nd Position: The nucleus of the cell migrates upward from the ventricular surface toward the pia, where its DNA is
copied
3rd Position: The nucleus (with diploid DNA) settles back into ventricular surface
4th Position: The cell retracts its arm from the pial surface
5th Position: The cell divides in 2 (in the ventricular zone)

After Vertical Cleavage: both daughter cells remain in the ventricular zone to divide again and again
Vertical cell division dominates early in development to expand the neural population

After Horizontal Cleavage: daughter cell lying farther from the ventricular surface migrates up to take its position
in the cortex  won’t divide again. The other daughter remains in the ventricular zone for more cell division. This
method dominates later development.

- Most neurons born between 5th week – 5th month of pregnancy; peak of 250,000 neurons per minute
- Once a daughter cell differentiates (commits to a neuronal fate) it won’t divide again

What Determines the Cell’s Fate?

- Cell fate is regulated by differences in gene expression during development
- Gene expression is regulated by transcription factors (proteins) and mRNA
- If transcription factors are unevenly distributed within a cell, then the cleavage plane can determine which
factors are passed onto the daughter cells
- Example: Notch1 and Numb are proteins in different poles of ventricular zone precursor cells. If the cell divides
vertically, they’re split symmetrically. If it divides horizontally Notch1 goes with migrating daughter, Numb
remains with cell that will divide. Notch1 activates the gene expression that makes the cell migrate from the
ventricular zone.

- Mature cortical cells can be classified as glia or neurons, etc

- Neural Stem Cells (precursor cells) can differentiate into multiple cell types
- Ultimate fate of the migrating daughter cell is determined by a combination of factors:
1. Age of the precursor cell
2. Position within the ventricular zone
3. Environment at the time of division

2. Cell Migration:
- Daughter cells migrate by slithering along radial glia from the ventricular zone to the pia
- The neuroblasts (immature neurons) follow the radial glia toward the surface of the brain
- Not all migrating cells follow this path; 1/3 wander horizontally to the cortex
- The cells then distribute themselves throughout the developing CNS
- The first cells to migrate away are destined to reside in the Subplate (eventually disappears)
- Those destined to become the cortex migrate next – cross Subplate and form Cortical Plate
- First cells in Cortical Plate are layer VI neurons, and then layer V, IV, etc
- The cortex is said to be assembled "inside out"

3. Cell Differentiation
- Differentiation: the process in which a cell takes on the appearance and characteristics of a neuron
- Begins as soon as precursor cells divide with uneven distribution of cell constituents
- Continues as neuroblast arrives in the cortical plate
- Layer VI neurons differentiate into pyramidal cells before layer II cells migrate into cortical plate

Differentiation of the Neuroblast  Neuron:

1. Appearance of neurites sprouting off the cell body
2. The neurites differentiate into axons and dendrites

Differentiation occurs even if the neuroblast is removed from the brain and placed in tissue culture – this means
that differentiation is programmed before the neuroblast arrives in the cortex.

Differentiation of Cortical Areas:

- Most cortical neurons are born in ventricular zone, migrate along radial glia, end up in cortical layers
- Cortical areas of the adult brain reflect an organization that is present in the ventricular zone
- The ventricular zone contains a cortical "protomap"
- The thalamus is important for specifying the pattern of cortical areas
- Subplate neurons attract the appropriate thalamic axons to different parts of the developing cortex
- Ex: LGN axons to the occipital cortex, VP nucleus axons to the parietal cortex, etc…
- Thalamic axons are area specific and initially innervate distinct populations in the subplate
- When the overlying cortical plate grows to a sufficient size, the axons invade the cortex
- Arrival of the thalamic axons causes the differentiation we recognize in the adult brain

Genesis of Connections:
- As neurons differentiate, they extend axons that must find appropriate targets
- Pathway selection: Retinal cell axon going to the correct location in the LGN travels down optic stalk, reaches
optic chiasm, must decide which of 2 optic tracts to continue along (depends on location in the retina of the
ganglion cell)
- Target selection: Choice of which thalamic nucleus to innervate (LGN obviously)
- Address selection: Must find the correct layer of the LGN
4. Synaptogenesis
- Growth cone: growing tip of a neurite; specialized to identify path for neurite elongation and hunts for target cells
to make synapses with
- Lamellipodia (edge of cone) with spikes called filopodia; probe the environment
- Filopodia take hold of the substrate on which it's growing  growth of neurite
- Growth cone comes in contact with its target; a synapse is formed

5. Neuronal Cell Death

- Entire populations of neurons are eliminated during pathway formation
- After axons reach targets and synapse formation begins, there’s a neuron/axon number decline
- Cells compete for trophic factors secreted by target cells; die without them
- Thus only cells wired up correctly survive – those without effective contacts die
- Nerve Growth Factors (NGF): trophic factor produced by targets of axons; injection of NGF antibodies into
newborn mice results in basal ganglia degeneration
- Programmed because cell death is a consequence of genetic instructions to self-destruct
- Apoptosis; neurons die by disassembly of neuron

6. Synapse Rearrangement:
- Synapses are re-arranged based on patterns of neural activity
- Patterns of activity control stability of synapses
- Hebb Hypothesis: segregation is thought to depend on a process of synaptic stabilization, where only retinal
terminals active at the same time as their postsynaptic LGN target neuron are retained
- When presynaptic axon is active and postsynaptic axon is strongly activated, then synapse formed by presynaptic
axon is strengthened – Cells that Fire together Wire together
- NMDA receptors play a role in determining what synapses survive
- Presynaptic activation  glutamate release  acts on postsynaptic NMDA receptors
- Highly correlated activity of synapses (necessary for synaptic enhancement) causes sufficient depolarization to
relieve Mg2+ block of NMDA receptor channels

C. Genetic and Environmental Mechanisms: Optic Tectum

- Both genetic and environmental factors influence organization of the brain
- All adult frogs have the same topographic point to point mapping of the retina onto the tectum
- Normally frogs have 2 eyes and each retina projects axons exclusively to the optic tectum
- Eye buds are reversed but axons still project exclusively to the optic tectum
- Chemoaffinity hypothesis: chemical markers on growing axons are matched with complementary chemical
markers on the optic tectum
- This suggests a Genetic basis for organization

- Eyes innervate different stripes called ocular dominance columns

- The input segregates into stripes that look like ocular dominance patterns
- If retinal activity is blocked, axons from 2 eyes become mingled; differences in activity can be used to
segregate inputs (Hebbian models)
- Reversing the retina screws up behavior forever, suggesting a genetic basis

- Environmental example?
- Blocking action potentials and NMDA receptors also blocks fine tuning – mapping to connections isn’t precise.
- Sensitivity of cortical cells to the eyes depends on action potentials and NMDA receptors

Ocular Dominance:
- A kitten was presented with visual stimulus and both left eye and right eye neurons responded
- After monocular deprivation of the right eye for hours, t was no longer able to evoke a response
- The majority of neurons in the visual cortex of a normal animal are driven binocularly
- Left eye open  synapses that drive postsynaptic cell are strengthened
- Right eye closed  neurons in visual cortex are not responsive to the deprived eye

AMPA/NMDA:
- Closing eye prevents image formation on the retina, and doesn’t correlate with a strong post-synaptic
response, so NMDA receptors aren’t strongly activated
- Modest Ca+ entry results in removal of AMPA receptors from visually deprived synapse
- Fewer AMPA receptors causes synapses to lose influence over cortical neuron responses