MPHY2002: Introduction to Biophysics

Chapter 7: Chemical Synapses

7.1 Introduction

Synapses are specialised locations on a neuron that allow for

signal transmission to adjacent cells. At a synapse, the
membrane of the signal-transmitting neuron (the pre-synaptic
neuron) comes into close apposition with the membrane of the
signal-receiving neuron (the post-synaptic cell). In a chemical
synapse, changes in the pre-synaptic membrane potential are
converted to an increase in the concentration of molecules called
neurotransmitters the region between the presynaptic and
postsynaptic membranes (the synaptic cleft), where they initiate
changes in the postsynaptic membrane potential. There is a
different type of synapse called an electrical synapse, in which
there is direct electrical coupling between the pre-synaptic and
post-synaptic membrane potentials. This chapter will focus on
chemical synapses.

The basic steps involved with signal transmission at a chemical

synapse are as follows:

1. The synaptic membrane is depolarised. This

depolarisation represents the signal to be transmitted to
the adjacent cell. Often, it takes the form of an action
potential.
2. Voltage-gated Ca2+ ion channels that are present in the
membrane open in response to the depolarisation, which
leads to an influx of Ca2+ from the extracellular space into
the synapse.
3. The increased concentration of Ca2+ causes vesicles
containing neurotransmitters to fuse with the synaptic
membrane and release their contents into synaptic cleft.
4. The released neurotransmitters undergo diffusion and
some of them bind to receptors in the postsynaptic cell
membrane.

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5. The binding of neurotransmitters to the receptors

initiates a transient change in the postsynaptic membrane
potential. Positive changes are called excitatory; negative
changes are called inhibitory.

Figure 1. Simplified schematic diagram of a synapse and

the processes taking upon arrival of a presynaptic action
potential. Source: M. Hennig, Modelling Synaptic
Transmission.

There are many different types of neurotransmitters that play

different roles in the body. An important neurotransmitter that
is involved with signalling from neurons to skeletal muscle cells
is acetylcholine (Ach). At the neuromuscular junction, Ach is an
excitatory neurotransmitter, so that its release results in a
positive change in the postsynaptic membrane potential. In the
central nervous system, two prominent neurotransmitters are γ-
Aminobutyric acid (GABA) and dopamine. In the mature CNS,
GABA functions primarily as an inhibitory neurotransmitter.
Dopamine can function as an excitatory or an inhibitory
neurotransmitter, depending on the type of receptor that is
present in the post-synaptic membrane.

A neurotransmitter receptor can be a part of a ligand-gated ion

channel. In this case, binding of a neurotransmitter molecule
(which acts as the ligand) to the receptor can directly cause a
change in the flow of ions through the channel, which in turn can
contribute to a change in the potential of the membrane in which
the channel is embedded. A neurotransmitter receptor can also
be distinct from ion channels, in which case binding of a
neurotransmitter molecule can exert its influence on nearby ion

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channels with a molecular chain of events called molecular signal

transduction mechanisms.

Neurotransmitters that have been released into the synaptic cleft

must be cleared rapidly to allow for new signals to be
transmitted by the synapse. This can take place with several
mechanisms: neurotransmitters can be recycled by the synapse
with re-uptake pumps embedded in the synaptic membrane,
they can be actively degraded by enzymes, and they can also
passively diffuse away from the synaptic cleft.

7.2 Miniature End-Plate Potentials (Synapses are Leaky)

The fundamental importance of synapses in information

processing has spurred a great deal of interest in their electrical
properties. One question of great interest is the extent to which
synapses are deterministic: how accurately do they transmit
signals?

Exploring the electrical properties of synapses is a challenging

experimental problem. It is particularly challenging for synapses
in the brain, which are typically extremely small and crowded
together in very large numbers. Synapses adjacent to motor
neurons (a neuron that initiates muscle movement) and skeletal
muscle cells are usually easier to isolate and study and this is the
context in which the first observations were made.

Some of the foundational experiments and biophysical modelling

work on synaptic transmission was performed at University
College London in the 1950s by Prof. Bernard Katz and his
colleagues. One experimental paradigm that was particularly
fruitful involved a dissected muscle and a piece of attached nerve
(typically several cm in length), with the measurement electrode
placed within a muscle fibre directly across from the synapse (a
region called the end plate). To record voltages from this
electrode, which are called end plate potentials (EPPs), a voltage
trace was swept across a cathode ray tube and the image was
photographed the image by keeping the camera open.

An important clue about the nature of synaptic transmission

came from the observation that even when a synapse adjacent to
a motor neuron is not electrically stimulated, there are still
transient changes of the end plate potential. These

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depolarisations are called spontaneous EPPs1. An example of

spontaneous EPP recordings is shown in Figure 2.

Figure 2. Examples of spontaneous end-plate potentials

(EPPs). The arrow points to the first spontaneous EPP in
recording (a). Adapted from Boyd and Martin. J. Physiol.
132, 61-73 (1956).

Figure 2 tells us that spontaneous EPPs do not occur at regular

intervals. If we assume that spontaneous EPPs occur randomly,
we can create a probability model for their occurrences in which
the length of the time intervals is a continuous random variable,
T. To obtain the PDF for T, we can start with the question: what
is the probability that there is no spontaneous EPP for a
particular length of time interval t? To answer this question, we
partition this time interval t into n sub-intervals of length t , as
shown in the figure below.

In our model, we assume that the probability that a spontaneous

EPP occurs in a particular interval is independent of the
probabilities that spontaneous EPPs occur in any other interval.
For very short time intervals, this probability is proportional to
t , so that we can write:

1
Another term for spontaneous EPPs, which was used by Prof. Bernard Katz, is miniature EPPs.

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Pr 1 spontaneous EPP in time interval t   t , (0.1)

for a positive real constant  . It follows that the probability that

there is no spontaneous EPP in a time interval t is equal to
1  t , and consequently the probability that there is no
spontaneous EPP in the entire time interval t, Pr T  t  , is given
by:

Pr T  t   1  t 
n
. (0.2)

Substituting the relationship nt  t into Equation, we obtain:

n
 t 
Pr T  t   1   (0.3)
 n
If we take the limit as t   so that n   , we obtain:

Pr T  t   e t . (0.4)

The PDF of T, p  t  , which tells us the probability that a

spontaneous EPP occurs between times t and t  t , can be
expressed as:

p T  t  Pr T  t  Pr spontaneous EPP in  t , t  t   (0.5)

Combining equations (0.1) and (0.4), we obtain:

p  t  t   t  e  t , (0.6)

so that

p  t    e  t . (0.7)

The distribution of Equation (0.7) is called the exponential

distribution.

7.3 Probability Model for Excitatory Post-Synaptic Potential

Amplitudes

One of the striking aspects of spontaneous EPPs is how similar

they are. While spontaneous EPPs occur at random times (as
their name suggests!), they have very similar time courses. An
example of a time course for one spontaneous EPP that was

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observed by Boyd and Martin at UCL in the 1950s is shown in

Figure 3.

Figure 3. Excitatory End-Plate Potential (EPP). Adapted

from I. A. Boyd and A.R. Martin, J. Physiol. (1956) 132, 74-
91.

The amplitude of an EPP can be defined as the maximum value of

this time course. A histogram created from multiple spontaneous
EPP amplitudes by Boyd and Martin is shown in Figure 4.

Figure 4. Histogram of spontaneous EPP amplitudes

observed experimentally (bars), with the Gaussian
function of best fit superimposed. Adapted from I. A. Boyd
and A.R. Martin, J. Physiol. (1956) 132, 74-91.

The similarity of the amplitudes of spontaneous EPPs led to the

quantal release hypothesis, which states that neurotransmitters

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are released by synapses in discrete amounts (quanta). We now

know that this hypothesis is correct: vesicles contain similar
amounts of neurotransmitters and they either fuse to the
synaptic membrane where they release all neurotransmitter
molecules (one quantum), or they do not fuse and consequently
they do not release any neurotransmitter molecules.

The quantal release hypothesis was tested by measuring changes

to the post-synaptic membrane potential when the synapse is
electrically stimulated, which are called end-plate potentials
(EPPs). In these experiments, the synapse is electrically
stimulated multiple times2 and the corresponding EPP is
recorded (each stimulation/recording pair referred to as a trial).

To determine whether the EPP measurements support the

quantal release hypothesis, a probability model is invaluable. For
this model, we can start with the following assumptions:

(a) vesicles behave independently of each other;

(b) when a synapse is electrically stimulated, the number of
released vesicles is a random number;
(c) there is a large number of vesicles in the synapse, but
when a synapse is depolarised, there is low probability
that any particular vesicle is released;
(d) the EPP can be considered as a sum of contributions from
individual vesicles.

To obtain the probability distribution for the number of released

vesicles, M, we denote the probability that a particular vesicle is
released when the synapse is stimulated is p and the total
number of vesicles in the synapse as n. It follows from
assumption (a) that the probability that m particular vesicles are
released when the synapse is stimulated is equal to p m , and
consequently that the probability that the remaining n  m
vesicles are not released is equal to 1  p 
nm
. Using the fact that
there are “n choose m” ways of choosing m vesicles from a total
of n vesicles, we can obtain the probability that exactly m
vesicles are released:

n
Pr  M  m | n vesicles     p m 1  p  .
nm
(0.8)
m

2
To be precise, stimulation was applied to the nerve connected to the synapse, which resulted in electrical
stimulation of the synapse.

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Equation (0.8) tells us that the number of released vesicles has a

binomial distribution with parameters n and p.

Provided that our assumptions about synaptic behaviour hold,

the binomial distribution of Equation (0.8) is exact. This
distribution is very inconvenient, however, because the
factorials in the “n choose m” term become unwieldy for large
values of n. It turns out that there is a very useful approximation:
when n is large and p is small [assumptions (b) and (c)], a
Binomial distribution can be well approximated by a Poisson
distribution:

 me
Pr  M  m   , (0.9)
m!
where   np . This approximation will be justified in Appendix A
of this chapter.

To obtain the relationship between the number of released

vesicles M and the EPP amplitude V , we make use of the
assumption that released vesicles contribute additively to EPPs
[assumption (d) above]. As a first attempt to express this
relationship, we could write:

V  M  EPP amplitude from one vesicle (0.10)

Equation (0.10) is unrealistic because it reflects the assumption

that each vesicle makes an equal contribution to the EPP
amplitude. In practice, there is continuous variation in their
contributions that can arise, for instance, from variation in the
amount of neurotransmitter molecules in each vesicle.
Therefore, in our probability model for the EPP amplitudes, we
need to include two sources of variability: the number of
released vesicles and the contribution of each vesicle to the EPP
amplitude. The contribution of one vesicle is given by the
conditional PDF p  V | M  1 , multiplied by the probability that
one vesicle is released, Pr  M  1 . Likewise, the contribution of
two vesicles is given by the conditional PDF p  V | M  2  ,
multiplied by the probability Pr  M  2  . When we include
contributions of all n vesicles, we can write:
n
p  V    p  V | M  m  Pr  M  m  . (0.11)
m 1

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Provided that n is very large and the mean number of released

vesicles is small, we can take the limit n   without having a
significant impact on the sum, to obtain a general expression for
our probability model:

p  V    p  V | M  m  Pr  M  m  . (0.12)
m 1

Between Equations (0.11) and (0.12), we have replaced the

proportionality symbol for a finite number of terms with an
equality symbol for an infinite number of terms.

The experimental observations shown in Figure 4 suggest that

the contribution of one vesicle to the EPP amplitude, which is
given by the conditional PDF p  V | M  1 , is well described by
a Gaussian PDF:

p  V | M  1  G   ,  2 

1    V    2  . (0.13)
 exp  
2 2  2 2

 
It is worth noting that Equation (0.13) is an empirical equation
that is obtained by inspection of data instead of a fundamental
physical law. However, the choice of a Gaussian is not completely
arbitrary either: from the Central limit theorem, we can expect
that many probability distributions encountered in nature can be
approximated as Gaussian.

How can we extrapolate from Equation (0.13) to cases where

more than one vesicle is released? By our assumption (d), the
EPP can be considered as the sum of EPPs that arise from
individual vesicles. It turns out that the sum of two independent
random variables with Gaussian PDFs G   ,  12  and G   ,  2 2 
also has a Gaussian PDF, with the same mean and the sum of the
variances: G   ,  12   2 2  . It follows that the contribution of m
vesicles to the EPP amplitude is given by the following Gaussian
PDF:

p  V | M  m   G   , m 2 

1    V    2  . (0.14)
 exp  
2 m 2  2m 2 
 

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Substituting Equations (0.9) and (0.14) into Equation (0.12), the

general expression for our probability model, we obtain an
explicit expression for the PDF of EPP amplitudes:


1    V    2   m e  
p  V    exp   . (0.15)
2 m 2  2m 2  m!
m 1
 

7.4 Comparisons with Experimental Data

How well does the probability model that we derived in the

previous section [Equation (0.15)] fit experimental data? To
determine the answer to this question we need to estimate the
values for the three free parameters in Equation (0.15): µ, , and
.

To estimate the free parameter  from the Poisson distribution,

we note that according to Equation (0.9), the probability that no
vesicles are released when the synapse is depolarised is:

Pr  M  0  e   . (0.16)

In their experiments, I. A. Boyd and A. R. Martin observed that

the EPP amplitude was zero in 18 out of the 200 trials [need to
define]. We can therefore estimate:

18
Pr  M  0   0.09 (0.17)
200
Using Equation (0.16), we can obtain our estimate:
e   0.09    ln  0.09   2.41 .

Based on the Poisson distribution of Equation (0.9), we can

calculate the probabilities that different numbers of vesicles are
released. For our estimate   2.41 , these probabilities are
plotted in Figure 5.

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Figure 5. Probability distribution for the number of

released vesicles at a synapse, based on a Poisson
distribution with parameter   2.41 .

To estimate the parameters µ and  in Equation (0.15), we can

examine histogram of spontaneous EPPs in Figure 4 (which we
assumed to occur from spontaneous releases of individual
vesicles). We can perform this estimate in two ways. The first
way is to use visual inspection – noting the µ, the approximate
centre of the distribution, occurs at 0.4 mV and that , the
approximate inflection point of the distribution, occurs around
0.1 mV. The second way is to perform a fit of the distribution,
from which we can obtain more accurate estimates:   0.4 mV
and   0.088 mV .

By plugging our estimates for µ and  into Equation (0.14), we

can obtain the conditional PDFs p  V | M  m  for different
numbers of released vesicles: m  1 , m  2 , and m  3 (Figure 6).

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Figure 6. Conditional PDFs p  V | M  m  of the EPP

amplitudes for m  1 , m  2 , and m  3 . Each distribution
was calculated from Equation (0.14), with   0.4 mV
and   0.088 mV .

By plugging our estimates for µ and  into Equation (0.15), we

can obtain the PDF p  V  , which is plotted in Figure 7.

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Figure 7. PDF for the EPP amplitudes (solid line), as

calculated with Equation (0.15). The conditional PDFs
p  V | M  m  for m  1 , m  2 , and m  3 are shown in
dashed lines.

From Figure 7, we can see that there are several peaks in the
PDF: an initial peak at 0.4 mV corresponding to stimulation trials
in which there is 1 released vesicle, a second peak around 0.8 mV
(2 released vesicles), and two small peaks at 1.2 and 1.6 mV (3
and 4 released vesicles, respectively). There are no peaks in the
PDF corresponding to 5 or more released vesicles due to the
increasing broadness of conditional PDFs p  V | M  m  as the
number of released vesicles m increases.

Boyd and Martin found that the probability model of Equation

(0.15) accounted for their data very well (Figure 8). These
results provided strong evidence in favour of the quantum
transmission model. In the years following these early
experiments, the quantum transmission model has been
validated with several different methods, including direct
observation with electron microscopy (Appendix B). This work
ultimately resulted in a Nobel Prize for Prof. Bernard Katz in

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1970.3 Research in synapses is still a field of intensive research

today.

Figure 8. End-plate potentials (EPPs) observed

experimentally (bars) that were induced by electrical
stimulation and the corresponding probability model
[solid line; calculated with Equation (0.15)]. Source: Boyd
and Martin. J. Physiol. 132, 74-91 (1956).

3
Prof. Katz’ Nobel Lecture can be found here:
http://www.nobelprize.org/nobel_prizes/medicine/laureates/1970/katz-lecture.pdf

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Appendix A: The Poisson Distribution as a limiting case of

the Binomial Distribution

Using the third assumption, we can convert equation x to a more

useful form. Equation x can be expanded as follows:

 n  n  1  n  m  1   n  m !   m   n  m
Pr  m vesicles released     1   (0.18)
m ! n  m  ! n  n
Taking all the terms without n to the left, and cancelling out the
 n  m ! terms, and re-arranging, we have:

  m   n  n  1  n  m  1          m 
n

Pr  m vesicles released        1     1   
 m!   nm   n    n  
, (0.19)
    n n  1 n  m  1          
m n m

     1  n   1  n  
 m!   n n n    
where the second line involves a re-arrangement of the second
square-bracketed term. In the limit that n   , the second and
fourth square-bracketed term both approach 1. For the limit of
the third square-bracketed term, we have:
n
 
lim 1    e  . (0.20)
n 
 n
 m 
Pr  m vesicles released   e (0.21)
m!
The probability distribution in Equation (0.21) is called a
Poisson distribution. The Poisson distribution has some lovely
properties, one of which is that it can be expressed in terms of
only one parameter, .

We have just seen that provided that n is large and p is small, a

binomial distribution can be approximated by Poisson
distribution.

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Appendix B: Visualising Synapses

Figure 9. Illustration showing the action of neurotransmitters in

the synaptic cleft. Vesicles containing the neurotransmitter
(green) move towards the pre-synaptic membrance where they
fuse with the cell membrane, releasing their contents into the
synaptic cleft. The neurotransmmitter molecules act on the post-
synaptic cell by binding to specific receptors on the cell surface
(purple). They can also be taken back up by the presynaptic cell
via other receptors (orange) for re-use. Credit: Arran Lewis,
Wellcome Images.

Figure 10. Image of axons making contact with muscle fibres

(green) at neuromuscular junctions (confocal light microscopy).
The neurotransmitter acetylcholine (red) sends the signals from
the nerve to makes the muscle contract. Credit: Tom
Gillingwater, Wellcome Images

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Figure 11. Illustration of the junction between an axon and a

muscle. Credit: Medical Art Service, Munich; Wellcome Images.

Figure 12. A light micrograph of teased striated muscle, showing

axons and motor end plates. The specimen was stained with
haemotoxylin and gold. Credit Spike Walker, Wellcome Images

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