Synaptic Transmission

Richard W. Keller, Jr., Ph.D.

Center for Neuropharmacology and Neuroscience
ME-751 KellerR@mail.amc.edu 2-5253

LECTURE OBJECTIVES
1.

To understand the overall concept of synaptic transmission.

2.

To understand the inputs into dendrites and the cell body.

3.

To analyze the differences between metabotropic and ionotropic

receptors.

4.

To understand the sequence of events in the presynaptic terminal

leading to neurotransmitter release.

5.

To list the methods of transmitter inactivation and discuss their

importance.

6.

To describe in the steps (synthesis, storage, release and inactivation

of acetylcholine) of neurotransmission at the neuromuscular junction.

7.

To analyze a disorder associated with the NMJ - Myasthenia Gravis.

MAJOR FEATURES OF A NEURON

There are many different types of neurons but they almost
all have similar major features.

Dendrites
(inputs)

Axon - AP Dr. Shin

(conduction of
message)
Synaptic
transmission

Cell Body
(metabolic
center)

Axon Terminals
(output)
Quick Review covered by Dr. Lindsley

What does this really look like????

Nerve Nerve
Connections
Artist John Allisons
Rendering of the Cortex

Neuromuscular
Junction

No Exam Material!

See Figure 11-36 Alberts, et al.

http://education.vetmed.vt.edu/Curriculum
/VM8054/Labs/Lab10/lab10.htm

MAJOR FEATURES OF A NEURON

Todays
Focus

Todays
Focus

Chemical Signal

Dendrites
(inputs)

5/68

Electrical Signal

Chemical Signal

Why these changes in message??

Cell Body
(metabolic
center)

Axon
(conduction of
message)

Axon Terminals
(output)

Major features common to almost all nerve cells.

1. Cell Body
a. Basic machinery of life. The cell body of the
neuron contains all the same organelles that are
found in all animals cells.
[nucleus, rough endoplasmic reticulum (ER),
smooth ER, Golgi apparatus, mitochondria - lots!]
b. The cell body functions as the metabolic center,
the site of protein synthesis & packaging, etc.
c. Diameter 5 - 60 um - small compared to most cells
d. Some inputs from other neurons - most via dendrites

Quick Review covered by Dr. Lindsley

Fig 1.3 Neuroscience; 2nd Ed.

Purves, et al 2001; Sinauer

THE CELL BODY

Nucleus

No Exam Material!

The cell body =

~0.1% of entire
cell volume!

Quick Review covered by Dr. Lindsley

2. Dendrites
a. Communication: neurons receive inputs (messages)
from adjacent cells to pass on to other cells. Many
of these inputs (afferent signals) are on
projections from the cell body called dendrites.
1. "antennas" of the neuron
2. branched in a "tree-like" fashion
b. Several dendrites - number and extent of the
branching varies dramatically
1. average neuron receives about 10,000
inputs in its dendritic tree
2. Purkinje cells - very large dendritic tree
frequently receiving 150,000 inputs
INPUTS = NEUROTRANSMITTERS interacting
with RECEPTORS (Ligand-gated Ion Channels!)

Inputs

A typical
Neuron has
about

10,000
inputs into
the dendrites
Thousands of
excitatory and
thousands of
inhibitory inputs!

Excite
Inhibit

Receptors

A cell potential exists across all parts of a neuron

including the cell body and dendrites.
The resting membrane potential is ~ -80 mV
From last lecture - primarily due to a K leak!

In fact, all living cells have membrane potentials!

so.... Whats the big deal about potentials in neurons?

Significant Changes in the cell potential result from NTs

interacting with & opening LIGAND-GATED Ion-Channels!

Really 2 types of
Neurotransmitter-Activated Receptors
1. Ligand-gated
Ion channels

Direct!

2.

Metabotropic
receptors

Indirect!

FAST!

2nd messengers

SLOW!

neurotransmitter = ligand
Fig 5.16 Neuroscience; 5th Ed.
Purves, et al 2012; Sinauer

Lots of Inhibitory and Excitatory Inputs

Some inputs
function to
"excite" the
cell, while
others tend
to "inhibit"
the cell.

At a cell
potential level
what does
excite mean?
"inhibit"?

Reality is a combination of Excitation & Inhibition

Excitation

Inhibition

Excitation & Inhibition

Initial
Resting pot.
-80 mv

-80 mV

Depolarizing
Less (-) more (+)
-50mv 0 mv
(+) current in
15/68

Hyperpolarizing
More (-)
-100mv
(+) current out
or (-) current in

-80 mV

Reality is a
combination of the
two effects.
Fig 12-16 Principles of Neural Science
4th Ed. Kandel, Schwartz & Jessell

What are the most common Inputs on

these Dendrites??
Of the 100 billion nerve cells in the CNS:
50-60% OF ALL NEURONS IN THE CNS USE
GLUTAMATE AS THEIR NEUROTRANSMITTER!
30-40% OF ALL NEURONS IN THE CNS USE
GABA AS THEIR NEUROTRANSMITTER!
So significant number of inputs on the dendrites
are likely via Glutamate and GABA.

GABA = Gamma-aminobutyric acid

In CNS Most Excitatory & Inhibitory Inputs are Glu & GABA

Glutamate = major excitatory NT

Glu

More
Positive;
depolarizing

Na+

Glutamate
Excitation

GABA = major inhibitory NT

Glu

GABA

Initial
Resting
pot.

-80 mv

Cl-

GABA

More
Negative;
hyperpolarizing

GABA
Inhibition

outside
Does xxx
produce
excitatory or
inhibitory
effects via
this receptor???

xxx

xxx

K+
inside
No Exam Material!

Self Study
You think about
this!

Activation of Receptors Results in Graded

Potentials in the Dendrites & Cell Body
Excitatory
Input

Apply Glutamate (Glu) into area with Glu receptors

Dendrite

ligand-gated ion channels..

signal decays over distance..

a graded potential !!
Adapted from Fig 2.3 Neuroscience;
5th Ed. Purves, et al 2012; Sinauer

B. There are thousands of excitatory and

inhibitory inputs on dendrites & the cell body
1. cellular decision has to be made to fire or
not to fire an action potential
2. potential is summed over area (spatial
summation) & time (temporal summation)
3. the trigger zone is located in the initial
segment of the axon; this is where the
cellular decision is made. [Adjacent to the
axon hillock region of the cell body.]
Initial segment - very
high density of voltagegated Na+ channels
axon

Axon Hillock
20/68

If the cell potential in the

initial segment is
sufficiently positive to
lead to the opening of
voltage-gated Na+
channels the cell fires
an action potential.

Action Potential moves down the Axon!

axon = neuron's output - the "wire" leading to where the message
will be delivered
Na+
Channel
= AP

All or None?

K+
Channel
= recovery
No Exam Material!

Covered by Dr. Shin and in Membrane Transport

ENERGY
REQUIRED!

Na+/K+ Pump

No Exam Material!

Covered in Membrane Transport

From Fig 4.11 Neuroscience;

5th Ed. Purves, et al 2012; Sinauer

CELL BODY
and
DENDRITES

AXON
AXON HILLOCK
threshold - fire?
spatial summation
temporal summation

TERMINAL FIELD
AXON Terminals
or boutons

VOLTAGE-GATED ION CHANNELS

ALL OR NONE ACTION POTENTIAL!
INFO CODED IN FREQUENCY
LIGAND-GATED ION CHANNELS
(Neurotransmitter-activated)
METABOTROPIC RECEPTORS that
modify ion channels
Excite - depolarize
Inhibit - hyperpolarize
GRADED POTENTIALS!

Examine what
happens here!

Synaptic Transmission
The big picture

Presynaptic terminal:
contains synaptic vesicles loaded
with neurotransmitter molecules
Lots of mitochondria (synaptic
activity consumes lots of energy)
in the terminal.

Synaptic cleft: 20-30 nm

Crisscrossed by thin filaments

Postsynaptic component:
contains intrinsic membrane
proteins (receptors) to which the
released transmitter can bind. This
can trigger a variety of
postsynaptic events. DENDRITE OR MUSCLE FIBER!!

Chapter 5 - Box 5A
Neuroscience; 5th Ed.
Purves, et al 2012; Sinauer

Synthesis of Neurotransmitters
THE NAME OF A NEURON COMES FROM THE
NEUROTRANSMITTER THAT IS RELEASED

can be up to 2 meters in human!

synthesis of
Neurotransmitters

synthesis of
Enzymes &
Vesicles
25/68

Figure 5.5 Neuroscience;

5th Ed. Purves, et al 2012;
Sinauer

The Synthesis, Packaging and

Secretion of Neurotransmitters
Biosynthetic
Enzymes
Storage Vesicles

Fig 5.3 Neuroscience; 5th Ed.

Purves, et al 2012; Sinauer

Synaptic Terminals / Synaptic Boutons -cont

An action potential (AP) moves down the axon and
invades the terminal area. This AP opens voltagedependent Na+ channels and then voltagedependent Ca+2 channels leading to an influx of
Ca+2.
This influx of Ca+2 is the trigger for exocytosis
which leads to the release of neurotransmitter.
In this process, vesicles fuse with the plasma
membrane of the terminal and dump their contents
into the cleft of the synapse.
Terminals conserve membrane by recycling it; after
release, vesicles are reformed and refilled.

EXOCYTOSIS - the simple version!

Docked
Vesicle
Plasma
membrane

Fig 5.14 Neuroscience; 5th Ed.

Purves, et al 2012; Sinauer

A REAL
VESICLE!
Takamori, et al.
Cell 127: 831-846

More of the
components
involved in
exocytosis
and then a
miracle
happens!
No Exam Material!

Fig 5.10 Neuroscience; 2nd Ed.

Purves, et al 2001; Sinauer

Role of Ca+2 in Synaptic Transmission

Control

Remove Ca+2 from Buffer!

Why this
delay?

2
2
?

30/68

Fig 5.11 Neuroscience; 5th Ed.

Purves, et al 2012; Sinauer

There are 2 major

types of release sites.

PROJECTION AREA
OR

TERMINAL FIELD

Those with AXON terminals

Acetylcholine
Glutamate
GABA

10,000
terminals

Those with AXON boutons

Dopamine
Norepinephrine
Serotonin

500,000
boutons

Associated with these two different fields are

2 types of synaptic contacts that release NTs.
1. Axon
Terminals

Classical
Neurotransmission
axon

2. Axon
Boutons

axon
Glutamate
GABA
(ACh - NMJ)

Extracellular
levels IMP!
Volume Transmission
Synaptic cleft
levels - IMP!

Catecholamines
& Serotonin

Removal of Neurotransmitters
Resetting the system for the next message!
Which drug blocks the removal of ACh at the NMJ and is a threat to life?
Nerve Gas loss of the ability to breath cant move diaphragm!!

Release must be terminated and the synapse

must be cleared of transmitter so that the next
message can produce its effect.

The mechanisms of NT removal used are:

1. Uptake or Reuptake
a. Reuptake in the cells that released the NT
b. Uptake into another cell

2. Enzymatic degradation
3. Simple diffusion away

ReUptake
into Boutons

MAO
metabolism

DAT

40%

AP

1 of the 500,000 boutons

DAT

80%

DA Uptake 1

DA Uptake 2

Transporters (Trans) are

protein carriers that
move molecules through
the plasma membrane

60%

DA

DA

DAT

DA

COMT
metabolism
20%

DA

ECF

DA recp

DA

DA
1

vesicular uptake
VMAT

DA recp

DA recp

Post Synaptic
Target Cell
35/68

Dopamine
Norepinephrine
Epinephrine
Serotonin

Other Cells
DA-term.ppt

DAT = Dopamine Transporter (Reuptake)

Uptake into
Astrocytes

(Astrocyte)
Major removal
Site! Glu & GABA
Transporters

Nerve Terminal

Glutamate
GABA
Fig 7.2 Fundamental Neuroscience;
2nd Ed. Squire, et al 2003; Academic Press

Removal of Neurotransmitters
1. Uptake
2. Enzymatic degradation
1. In the ECF
2. In other cells
3. In the releasing terminal / bouton
3. Simple diffusion away

Enzymatic
Degradation

NeuroMuscular
Junction

ACh
Choline
+ acetate
ACh
Histamine
Thanvi & Lo - Postgrad Med J 2004;80:690-700

Acetylcholinesterase

Removal of Neurotransmitters
1. Uptake
2. Enzymatic degradation
3. Simple diffusion away

Diffusion

Fig 7.2 Fundamental

Neuroscience;
2nd Ed. Squire, et al 2003;
Academic Press

All NTs to
some extent!

Recovery from Depolarization of Release

(Recovery of the Terminal)

Fig 4.9 Neuroscience; 5th Ed.

Purves, et al 2012; Sinauer

Events from Neurotransmitter Release

to Postsynaptic Excitation or Inhibition
Neurotransmitter
Release

Receptor Binding in Post synaptic cell

Metabotrophic
Receptor - Second
Messengers
Does binding to a
receptor play a role
in NT removal?

Ligand-gated Ion channel

opening or closing

Postsynaptic cells membrane potential

change - excited or inhibited

Postsynaptic
Presynaptic
autoreceptors

Locations of
Receptors

(Feedback enough NT! )

Presynaptic
heteroreceptors
Nt = neurotransmitter
Not much
evidence
for these
hetero receptors
at NMJ.

Nt

Nt

Nt2

Nt

Nt3

Nt

Nt2
Nt

Nt3

Nt

Nt
Nt

There are
presynaptic
autoreceptors
at NMJ.

Ion channel
Metabotropic receptor
45/68

The Combined Effect of

Neurotransmitters and their Receptors
Many different kinds of chemical messages
Fast vs slow Some neurotransmitters (NTs) work very
quickly and are rapidly removed (ACh at NMJ) vs
others that are more neuromodulators (DA in brain)
Receptors also add variability. The same NT can be
Excitatory at one receptor and Inhibitory at another.
There are receptors with different affinities for the
same NT; so low levels of NT can stimulate one set of
receptors and high levels can stimulate additional
receptors.

Same NT Different Responses at Different

Acetylcholine Receptors

Excitatory

Inhibitory

Nicotinic
cholinergic
ion channel
receptor

Muscarinic
cholinergic
metabotropic
receptor

see animation of inhibition @ http://www.blackwellpublishing.com/matthews/neurotrans.html

Ion Channel Receptors

Many Different
Receptors Respond
to Different levels of
Neurotransmitter

Metabotropic Receptors
Ion Channel vs
Metabotropic
Excitatory vs
Inhibitory
Some respond to low NT
levels others respond
to high NT levels
Fig 6.3 & 6.4 Neuroscience; 5th Ed.
Purves, et al 2012; Sinauer

Do Not Memorize

The Neuromuscular Junction (NMJ)

Specialized chemical synapse between a motor neuron
and striated muscle fibers. The NT used is acetylcholine
(ACh)
Synthesis: from acetyl CoA and choline by choline-acetyl
transferase (CAT) in the terminal
Storage & release: Uses mechanisms described previously
Receptors: NMJ uses nicotinic acetylcholine receptors.
These are ligand-gated Na+ (some K+) ion channels that will
generate a depolarization when bound to ACh. This
depolarization is called the endplate potential (EPP).
Inactivation: ACh is inactivated enzymatically. The synaptic
cleft in NMJ contains acetylcholinesterase (ACE) which
breaks ACh into acetyl CoA and choline. Choline is brought
into the terminal via a transporter for reuse in synthesis.
50/68

The Neuromuscular Junction (NMJ):

NMJ

http://demo.classontheweb.com

http://education.vetmed.vt.edu/Curriculum
/VM8054/Labs/Lab10/lab10.htm

Surface Area of a Presynaptic Terminal

The Neuromuscular Junction (NMJ)

A lot of what we
know was first
worked out at the
NMJ!!

CNS Synapse

Saladin Anatomy and Physiology:

The Unity of Form and Function 1996
McGraw-Hill

2,000-6,000 um2

Not to
Scale!

2-10 um2

NMJ
Freeze
Fracture
Slam the tissue
against a copper
plate frozen in
liquid nitrogen
and then break it
apart.

Do Not Memorize

Freeze Fracture

EM

Unstimulated

Stimulated

John Heuser = http://www.heuserlab.wustl.edu/v2.0/index.shtml

Heuser, et al. 1979

J.Cell.Biol 81 275

Recordings in the junction reveal graded local potential

changes (EPP) before an Action Potential is produced.

Stimulated ACh release ~150 MEPPs (vesicles) = EPP

An EPP involves ligand-gated ion channels if the membrane
potential change is sufficient (exceeds the voltage-gated Na+
channel threshold) then an AP that involves voltage-gated ion
channels occurs in the muscle fiber.

The discovery of Miniature Endplate Potentials

minis or MEPPs
Membrane potential
of muscle fiber

spontaneous 0.5mV
changes in potential
(single vesicle leak)

EPP

Nerve stimulation
Bernard Katz
1950s

evoked change with

stimulation (APstimulated release of
~150 vesicles of ACh)

Skeletal Muscle Action Potential

in the muscle, away from the neuromuscular junction
the AP is again all-or-nothing (no EPP!)
EPP is playing
a role similar to
which region in
the neuron?

When the depolarization produced by the EPP is sufficient to activate

voltage-gated Na+ channels, an action potential is generated in the
muscle fiber.
This action potential results in opening of voltage-gated Ca+2 channels
located both in the cell membrane and in endoplasmic reticulum membranes
of the muscle fiber. The consequence is a massive entry of Ca+2 into the
cytoplasm, both from outside the cell and from intracellular storage sites,
which results in contraction of the muscle fiber.

Curare can limit the size of the EPP and thus block the
initiation of the Action Potential.
Curare (a cholinergic nicotinic antagonist) binds to
nicotinic cholinergic receptors but produces no
effects. If it is added to the bath, it blocks the binding
and effects of ACh at these receptors reducing the
EPP below the voltage-gated Na+ channel threshold
thus preventing the AP.

French physiologist Claude Bernard mid 1800s

60/68

Myasthenia Gravis
MG is an autoimmune disease that disrupts transmission at the
NMJ.
The body forms antibodies to nicotinic ACh receptors
- destroys some receptors and blocks others
There are less functional receptors and thus weak muscles.

Receptors
blocked by
antibodies!

Y Y

Myasthenia Gravis cont

MG typically affects the cranial muscles first
weak neck muscles cause the head to fall forward or
backward
drooping eyelids (ptosis)
inability to move the eyes side to side
double vision (diplopia)
difficulty speaking (dysarthria)
difficulty swallowing (dysphagia)

Weakening of the
Muscles!!!

Myasthenia Gravis cont

One of the most treatable neuromuscular disorders!
Administer acetylcholinesterase blockers
such as neostigmine or pyridostigmine
(+ immunosuppressive drugs)

This reduces the ACh destruction by AChE & increases the

accumulation of ACh at the neuromuscular junction. Hence
there is more ACh to interact the remaining receptors.

Before
Neostigmine

After
Neostigmine

Walker MB (1934).
Treatment of myasthenia
gravis with physostigmine.
Lancet 1:1200-1201.

Sample Questions:
1. Neurotransmitters (ligands) activate ion channels in the dendrites resulting in
membrane potential changes that are described as:
a.
b.
c.
d.
e.

all or none
negative
graded
positive
integrated

2. A rabbit shot with a curare-tipped arrow would quickly lose its ability to run
away, because at the neuromuscular junction
a.
b.
c.
d.
e.

the breakdown of acetylcholine would be inhibited.

the acetylcholine receptors would be blocked.
acetylcholine could no longer be released from the presynaptic terminals.
the voltage-dependent sodium channels would be blocked.
synthesis of acetylcholine from choline would be inhibited.

The End!

cb