Structure & Function of Neurons

10:00 on 9/15/2015
Dr. Lindsley
MS-539, lindslt@mail.amc.edu
Lecture Objectives: Introduction to the histology and cell biology of neurons,
emphasizing the special features of neurons that distinguish them from other cell
types, esp. axon transport. Using the example of a simple spinal reflex circuit,
students learn to integrate their understanding of neuronal structure and direction of
information flow to the gross anatomy of spinal nerves.
Student Learning Objectives:
1. Describe important histological characteristics of neurons, nerves and nervous
tissue.
2. Describe key features of the 3 kinetic components of axon transport (fast
anterograde, fast retrograde and slow anterograde).
3. Relate what you know about the structure of neurons and direction of information
flow to the anatomical organization of the spinal nerve.
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What is special about neurons compared to other cell types?

1. Highly polarized functional compartments
The cell body, dendrites, and axon are distinct compartments that differ in
structure and function.
Each compartment has subcompartments whose molecular characteristics
provide for specialized functions.
Very high level of transcription and protein synthesis, which occur almost entirely in
the cell bodies and dendrites (nissl substance).
Intracellular materials must be transported to regions far away from the site of
synthesis (axon transport). Axon transport is regulated by activity and myelination.
2. Excitability & Conductivity Cell-to-Cell Communication (Dr. Shins lectures)
All cells have different ion concentrations inside and outside the plasma membrane.
Muscle cells and neurons use this differential ion concentration to generate
electrical signals; both cell types respond to physical or chemical stimuli with a
change in the electrical potential across their plasma membrane.
The unique event in neurons occurs in axons: This electrical signal propagates
over long distances as a self-regenerating, non-decremental and uni-directional
wave (from the cell body to the synaptic terminals ) called an action potential.

What do you already know about the structure of neurons?

Q: How many cells are depicted in this drawing?

Add Labels: dendrite, neuronal cell body, nucleus, axon, Schwann cell nucleus, myelin
sheath or internode, Node of Ranvier, axon terminals.
Q: What is the direction of information flow in neurons?

There is a large diversity of neurons in the nervous system more

than any other organ system or cell type.
They all have regions for input, integration, conduction & output.
*

Modified from Kandel et al. 4th Edition. Fig 2.8

NO dendrites
NO synapses on soma
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Input from special
sensory organs

Classification of Neurons
There are several hundred different types of neurons that may be
classified by one or more of the following characteristics:
shape; multipolar, bipolar, unipolar, pseudounipolar, spiny,
pyramidal, etc.
modality; ex. sensory or motor

Details at 10am 9/17

whether they are connected to local targets (interneuron) or

distant targets (projection neuron)
neurotransmitter they release; ex. GABAergic, glutamatergic,
acetylcholinergic, etc.
Electrophysiological firing patterns; ex. bursting, regular spiking,
silent, etc.
various other characteristics

Ultrastructure of Neurons
Cell body input and integration
Postsynaptic regions (axo-somatic synapses)
Nissl substance
Axon hillock
Dendrites input and integration
Mixed polarity of microtubules
Postsynaptic regions (axo-dendritic synapses)
Dendritic spines
Axons conduction and output
Uniform polarity of microtubules
Initial segment
Node of Ranvier
Presynaptic regions (with output to neurons and other effector targets)

Histology of Dendrites

Input region - Primary target for synaptic

input from other neurons. Electrochemical
signals travel along dendrites to the soma.
They extend from the soma and taper in
diameter.
Length, thickness and pattern of branching is
highly variable.
Shape influences electrical properties and
surface area correlates with # of synapses.
Cytoplasm composition is similar to soma
(next slide), except that the golgi apparatus
and Nissl bodies are only in the proximal part
of the dendrites.
Unique features
Dendritic spines are tiny mushroom-shaped
protrusions on the dendrites of some neurons.
They increase the receptive surface area for
synapses, esp. excitatory input. Thought to be
important in neuronal plasticity & learning.
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Somatodendritic Region = combined input region; dendrites and soma of the cell.

Dendritic Spines: Ultrastructure

K. Harris 2004

Spines on a dendrite.
Reconstructed from serial EM

Most spines are mushroom-shaped; spine neck and head.

Spines receive synaptic input, primarily on their head region
(i.e. spines are postsynaptic).
Spine shape changes can occur rapidly in response to
synaptic activity and other conditions.
Spine shape influences electrophysiological properties.
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The structure of axo-spinous synapses in the CNS: LM and EM

C. and D. abbreviations:

A. Multipolar neuron with spiny dendrites.

B. Dendritic shaft (orange) and spines (green).

pre
ax
s.v.
PSD
dend

presynaptic
axon
synaptic vesicle
postsynaptic density
dendrite

Dendritic Spines: Plasticity

Before (left) and after (right) repetitive synaptic stimulation at

a synapse onto the spine indicated by the arrow.

Spine number and shape is modified by synaptic activity.

- The spine neck is rich in actin (shape change modifies electrical properties)
- local mRNA translation (activity-dependent changes in protein expression)

Spine number and shape may correlate with neural function and can be modulated by a
variety of conditions:
- Spine number is lower in conditions marked by cognitive impairment, ex. Autism spectrum
disorders, Downs syndrome, depression, chronic stress, normal aging.
- Spine number is increased by environmental enrichment and estrogen replacement.

More about spines in learning & memory in Nervous System I theme.

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Histology of the cell body (aka soma, perikaryon)

Input and integration; receives and summates
synaptic input.
metabolic and genetic center of the cell
nucleus (often has prominent nucleoli)
o post-mitotic
o lots of euchromatin (active transcription)
lots of sER, mitochondria, golgi

Unique features
Nissl substance a neuron-specific
arrangement of rER and polysomes that is
distributed throughout the cell body cytoplasm
(except in the axon hillock).
Axon hillock a region of cytoplasm in the cell
body that is adjacent to the axon and is devoid of
Nissl. Thought to be the trigger zone for the
action potential.
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Nissl substance is unique to neurons. This is an EM of Nissl.

Nissl substance is a unique
arrangement of cytoplasmic, free
polysomal rosettes (one rosette is
boxed) interspersed between rows of
rough endoplasmic reticulum (RER)
studded with membrane-bound
ribosomes (arrow).

Squire et al., 2nd Edition, Fundamental Neuroscience

Nissl bodies refers to the clumped

appearance of nissl substance in
stained sections observed by light
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microscopy (next slide)

LM showing Nissl stained sections of spinal cord grey matter.

Nissl bodies are the dark purple clumps in the cytoplasm. Recognize the
axon hillock region of the cell body cytoplasm by the absence of Nissl.

Neuronal
cell bodies

Axon hillock
The axon hillock is located within the cell body adjacent to where the axon
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emerges. Axons have NO Nissl substance.

Histology of Axons
+-

++

+
+

The conducting and secretory output regions.

Thinner than the dendrites; do not taper
Length and branching vary greatly.
Unique features
Axons have no Nissl (low protein synthesis).
Initial segment of the axon is adjacent to the
axon hillock in the soma; it has a high density of
Na2+ channels for initiating the action potential.
May be myelinated (shown) or unmyelinated.
Myelin is a multilayered wrapping of membrane
produced by glial cells. (Dr. Mongins lecture)
Internodal regions segments wrapped in myelin
Nodes of Ranvier unmyelinated regions
between adjacent internodal regions; nodes
have high density of Na2+ channels.
Axons have presynaptic specializations (at
terminals or nodal regions) where chemical
neurotransmitters are released; these terminals
or boutons are the output region (Dr. Keller).
All the microtubules have their plus-ends
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pointing away from the cell body.

Histology of Synapses

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What do you already know about the structure of synapses?

dendrite

cell body
(soma)

Node of
Ranvier

nucleus

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Neurons form synapses with other neurons.

Types of synapses can be classified by their locations.

Axospinous

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Histology of chemical signal transmission between neurons.

The axo-dendritic synapse occurs between the axon on one neuron (presynaptic)
and the dendrite of another neuron (postsynaptic).
Notice the many synaptic vesicles and mitochondria in the presynaptic terminal.
Notice the synaptic cleft (thickened pre- and post-synaptic membrane).
Notice the thickened and complex structure of the postsynaptic density.
Many synapses are surrounded by glial cells (tripartite synapse) Dr. Mongin
More about the cellular physiology of synaptic transmission from Dr. Keller.

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Neurons also form synapses with muscle.

The neuromuscular junction (NMJ) is a synapse between the axon of a
multipolar motor neuron (presynaptic) and a muscle cell (postsynaptic).
(More about NMJs from Dr. Mazurkiewicz).

axon of motor neuron

presynaptic
terminal

muscle

LM

muscle cell

EM
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Mechanisms of Axon Transport

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General Features of Axon Transport

Fast transport of membranous organelles
both directions:
anterograde (away from the cell body)
and retrograde (toward the cell body)
Slow transport of cytosolic proteins and cytoskeletal proteins
- anterograde direction only
Three Kinetic Components of Axon Transport
1. Fast anterograde
2. Fast retrograde
3. Slow anterograde (component a, component b)

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All axon transport components share the following biochemical and

molecular characteristics.
1. axon transport requires microtubules and motor proteins.
2. axon transport depends on oxidative metabolism (requires ATP)
3. axon transport does NOT require protein synthesis
4. axon transport is independent of the cell body

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Properties of microtubules (MT) in non-neuronal and neuronal cells

tubulin
- end (slow growing)

tubulin
+ end (fast growing)

Review: MT in non-neuronal cells

Microtubules in neurons

Microtubules are hollow tubes with walls

formed by 1214 protofilaments. Each
protofilament consists of a series of - and tubulin dimers organized in a polar fashion,
giving the microtubule a plus (fast growing)
end and a minus (slow growing) end.
Microtubules are approximately 24 nm in
diameter and may be more than 100 m in
length. Various polypeptides called
microtubule-associated proteins (MAPs) are
typically associated with the surface of the
microtubule. They regulate assembly,
stiffness, spacing and stability of the
microtubules. Microtubles in most cell types in

- Lots of tubulin ~ 10% of brain protein!

interphase are dynamic and unstable.

- are stabilized by microtubule-associated

proteins (MAPs).
2 important groups of neuronal MAPs:
Taus: axonal, developmentally-regulated
High MW MAPs: MAP-2 is dendritic, MAP-1c
is a cytoplasmic motor protein
- phosphorylated b-tubulin is neuron-specific
v In axons, microtubules have their plus ends
distal (pointing away) from the cell body
v In dendrites, microtubules have mixed
polarity; proximal dendritic microtubules have
either plus end or minus end distal; only the
most distal dendritic microtubules have plus
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ends distal (like axons).
Squire et al., 2nd Edition, Fundamental Neuroscience

Motor proteins involved in axon transport: Kinesins and Dynein

Kinesins
- 2 globular heads (motor domain) contain ATPase and microtubule-binding domains
- Binding to microtubules (MTs) activates the ATPase.
- tail domain (or stalk) interacts with cargo (e.g. membrane-bound organelles, MBO.)
- The mammalian brain contains a neuron-specific form of kinesin.
Dynein
- 2 globular heads (motor domain) contain ATPase and microtubule-binding domains
- Binding to microtubules (MTs) activates the ATPase.
- huge protein complex of ~1.5 megadaltons
- associated with dynactin (multiple proteins), regulates dynein cargo binding and activity.

Motor domain:
-ATPase
-MT-binding

Tail domain:
- MBO

Neuron-specific
Kinesin

Dynein

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Squire et al., 2nd Edition, Fundamental Neuroscience

Axon transport provides a mechanism for transporting material

between compartments in the axon.
1. Fast anterograde:
- moves material away from the cell body,
toward the + end of microtubules
- moves membranous organelles
e.g. synaptic vesicles, mitochondria
- motors are kinesins
- rate of ~200-400 mm/day

- ends
2. Fast retrograde:
- moves material toward the cell body,
toward the end of microtubules
- moves larger membrane-bound vesicles
e.g. prelysosomal, NGF and NGF-R
- motors are cytoplasmic dyneins
- rate of ~100-200 mm/day

+ ends

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3.

Slow axonal transport

(anterograde only; ave. 0.2 m/sec)

- moves material away from cell body

component a moves MT, NF and associated proteins
transported in assembled form
component b
moves variety of cytoplasmic proteins,
including actin, mRNA, and enzymes (e.g.
for neurotransmitter synthesis)
- motor may be a cytoplasmic dynein*
- two components:
The MTs being moved are NOT bound to the cargo
domain. Instead, the dynein is bound to actin near the
plasma membrane and unable to move. The motor
domain walking toward the (-)end displaces the MTs
toward the terminal (anterograde direction).

- ends

+ ends

*Q:

How can dynein that only carries cargo along

MTs in the direction of their minus ends, be a motor
for both retrograde AND anterograde transport?

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Axon transport is important in the pathogenesis of neurologic infectious and

neurodegenerative diseases. A few examples
Rabies virus: Viral particles gain access to the nervous system by traveling in motor neurons
by retrograde transport from the muscle to the cell body in the spinal cord.
Recent studies suggest that an interaction
between the rabies virus and the dynein light chain links the
virus RNP to the host cell retrograde transport system,
thereby facilitating axonal virus transport into the CNS.
(Jacob et al., J. Virology 2000; Raux et al., J. Virology 2000.)

Charcot-Marie-Tooth type 2 (CMT2): Charcot-Marie-Tooth disease is a group of progressive

disorders that affect the peripheral nerves, those connecting the brain and spinal cord to
muscles as well as those that detect sensations such as touch, pain, heat, and sound. It is the
most common inherited disorder that involves the peripheral nerves, and affects an estimated 1
in 3,300 people worldwide. Although the pathogenic mechanisms are still under investigation,
evidence suggests that abnormal mitochondrial transport may contribute to the selective
susceptibility of the longest axons, in which proper localization of mitochondria is critical for
axonal and synaptic function (Baloh et al., J. Neurosci. 2007).
Familial Alzheimers Disease (FAD): In a transgenic mouse model of FAD, defects in
anterograde fast axonal transport may contribute to reduction in the supply of neurotrophin
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receptors and other components of the presynaptic specialization, resulting in compromised
neuron function. (Lazarov et al., J Neurosci. 2007)

Tract tracing methods take advantage of axon transport to trace

connections in live, intact cells.
Anterograde tracing: Injection of tracer* into the region containing neuronal cell bodies
is followed by a period of time to allow anterograde transport of the label to distal axons.
Labeled tissue is then fixed and sectioned for histochemistry to visualize the location of
the labeling. * BDA (biotinylated dextran amine) or PHA-L (phytohemaglutinin-L)

Retrograde
axonal
transport

Retrograde tracing: Injection of tracer# into the region containing synaptic terminals is
followed by a period of time to allow retrograde transport of the label to the cell body.
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# HRP (horseradish peroxidase) or FluoroGold

Transneuronal labeling. Anterograde tracing may lead to the release of the label at
terminals and its uptake into synaptic target neurons. This permits labeling of
pathways involving multiple neurons.

Retrograde
axonal
transport

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Neurons in Spinal Nerves

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What is the difference between neurons, neural cells, & nervous tissue?
Neural CELLS are various types or classes of:
neurons (this lecture) specialized for intercellular communication
glia (Dr. Mongin) wide variety of nervous system functions

Nervous TISSUE refers to structures that contain neural cells along with other cells
and constituents. For example,
nerves in PNS, including cranial nerves
Dorsal root ganglia (aka sensory ganglia) in PNS
Autonomic ganglia in PNS
Grey and/or white matter of brain and spinal cord in CNS (aka neuropil)

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Know the difference between a neuron and a nerve.

A neuron is a single cell. A nerve is a type of nervous tissue in the PNS.
Nerves contain a bundle of axons (red) with variable amounts of myelin
(yellow), the cell bodies of other cell types (ex. Schwann cells, connective
tissue cells, the cells of blood vessels, and extracellular connective tissue
material. There are no neuronal cell bodies in nerves.

http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab9/Lab9.h
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tm

Begin using correct terms for nervous tissue in the PNS and CNS.

In the PNS (nerves and peripheral ganglia):

A bundle of axons is a nerve, root, trunk or ramus.
A cluster of neuronal cell bodies in the PNS is a ganglion.
In the CNS (brain, spinal cord and neural part of retina):
A bundle of axons is white matter and may be called a tract, column,
brachium, fasiculus, funiculus, or lemniscus.
A cluster of neuronal cell bodies is grey matter called a nucleus.
A layer or sheet of neuronal cell bodies is grey matter called a lamina or
cortex.
There are no nerves in the CNS. There are no collagen fibers between axons
in white matter.
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Formative Assessment Questions

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Recall the anatomy of Dr. Martinos typical Spinal Nerves

Locate:
Spinal cord in cross section
Gray matter (dorsal & ventral horns)
Dorsal and Ventral Roots (Nerves)
Dorsal Root Ganglia
Nerves
- typical spinal nerves
- rami
- nerves
What is the direction of information35flow?
From Noback et al., The Human Nervous System 6th Ed 2012, Humana.

Flow of information between PNS and CNS: A simple neural circuit

-- To other regions of the CNS
via white matter tract.

3 neurons participate in the circuit shown: sensory afferent neuron (red),

interneuron (blue) and motor efferent neuron (purple).
What is the location of the cell body of each type of neuron? Where are the synapses?
What part(s) of each neuron is in the CNS and what part(s) is in the PNS?
Locate the ventral roots, dorsal roots and dorsal root ganglia.
Locate the spinal nerves. What is the cellular composition of these nerves?36

List of textbook references:

Cui, Atlas of Histology, 1st Edition, Lippincott, Williams & Wilkins (Chapter 7; pgs 115-133)
Kandel, Schwartz & Jessell, Principles of Neural Science, 4th Edition. Elsevier
Kierszenbaum, Histology and Cell Biology: An Introduction to Pathology (2nd Edition). Elsevier
Paulina textbook
Purves, et al., Neuroscience, 4th Edition. Sinauer
Squire, et al., Fundamental Neuroscience, 2nd Edition.

Recommended for review:

Henrikson, Megargel & Murphy, Histology Laboratory: An Interactive Review, CD Version 2.0

Just for Fun

An animation of axonal transport called, The inner life of the cell
http://www.studiodaily.com/main/searchlist/6850.html
And with narration
http://multimedia.mcb.harvard.edu/media.html
Harvards Department of Molecular & Cellular Biology
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