MetaNeuron Manual

Eric A. Newman
Department of Neuroscience
University of Minnesota
MetaNeuron © 2002 Regents of the University of Minnesota. Eric A. Newman,
Department of Neuroscience. All Rights Reserved. For educational purposes only. Do
not copy or reproduce without permission. Contact Eric A. Newman at ean@umn.edu.

MetaNeuron was created and written by Eric A. Newman and Mark H. Newman. It was
inspired by Jerome Y. Lettvin’s transistor model of the axon, the MetaMembron. The
computer code was written by Mark H. Newman.

Version 1.03

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TABLE OF CONTENTS
Copyright notice 2
Overview of operating instructions 4
Introduction 6
Operation of MetaNeuron 7
Lesson 1, resting membrane potential 10
Student exercises 10
Lesson 2, membrane time constant 12
Student exercises 13
Lesson 3, axon action potential 15
Student exercises 16
Lesson 4, axon voltage clamp 20
Student exercises 21
Lesson 5, synaptic potential and current 24
Student exercises 24

ILLUSTRATIONS
Figure 1. Membrane time constant 13
Figure 2. Temporal summation 14
Figure 3. Action potential generation 16
Figure 4. Refractory period 19
Figure 5. Voltage clamp currents 21
Figure 6. Recovery from Na+ channel inactivation 23
Figure 7. Time course of recovery from inactivation 23
Figure 8. Reversal of an excitatory postsynaptic potential 25

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OVERVIEW OF OPERATING INSTRUCTIONS
These instructions can be viewed in the MetaNeuron program by clicking the About
button at the right of the screen.

MACINTOSH VERSION: Maximize the program window after opening.

CHOOSE A LESSON
Click on one of the five tabs at the top of the screen to select a lesson. Each lesson runs
independently; changing parameters in one lesson will not affect the operation of other
lessons.

CHANGE PARAMETER VALUES

Parameter values can be changed in several ways:
1) Click on the up or down arrow buttons to the right of a parameter box to increment
or decrement the value by 1. Pressing the "Shift" key while clicking a button increments
the value by 10. Pressing the "Ctrl" key while clicking a button increments values by 0.1.
2) Click on a parameter box. Keeping the mouse button pressed, drag the mouse.
Moving the mouse up or to the right increases the value by 1. Moving the mouse down
or to the left decreases the value. As in method “1”, the “Shift” and Ctrl” keys change the
increments to 10 and 0.1, respectively.
3) Click on the number in a parameter box, type in a new value, hit "Enter".

MACINTOSH VERSION: The “Shift” and Ctrl” key options and dragging the mouse
(method 2) do not work on Macintosh computers.

CURSOR
The X and Y values of points on the graph can be determined by moving the mouse over
the graph and clicking the left mouse button. The values, in units appropriate for the
yellow trace, are given in the upper right corner of the program window. The currently
selected point on the graph is indicated by crosshairs when the “Show Cursor” box is

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checked. Displayed X and Y values are confined to points along the yellow trace when
the “Trace” box is checked.

FAMILY OF CURVES
A family of curves can be generated by checking the “Keep Previous Graphs” box.
Graphs are retained as long as the box remains checked. A simple way to generate a
family of curves is to check the “Keep Previous Graphs” box, hold down the Shift key (to
increment in steps of 10), left click on a parameter box (e.g. stimulus amplitude) and
move the mouse to the right while keeping the left mouse button pressed.

DEFAULT VALUES
All parameters in a lesson can be reset to default values by clicking the “Reset to
Defaults” button.

MetaNeuron represents 1 cm2 of neuronal membrane. The membrane has a capacitance

of 1 µF.

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INTRODUCTION
MetaNeuron is a computer program that models the basic electrical properties of
neurons and axons. The program is intended for the beginning neuroscience student and
requires no prior experience with computer simulations.
Different aspects of neuronal behavior are highlighted in the five lessons
presented in MetaNeuron. The first two lessons, Membrane Potential and Membrane
Time Constant, illustrate the passive properties of neuronal membranes. The third and
fourth lessons, Axon Action Potential and Axon Voltage Clamp, demonstrate how
voltage- and time-dependent ionic conductances contribute to the generation of the action
potential in the axon. The fifth lesson, Synaptic Potential and Current, illustrates how
synaptic potentials are generated through the activation of ionotropic neurotransmitter
receptors.
The lessons in MetaNeuron do not attempt to model the full complexity of
neuronal behavior. The simulations simplify neuronal properties, highlighting the basic
principles of neuronal function.
Other, comprehensive simulations of the neuron are available to students who
wish to investigate neuronal behavior in more detail. One simulation in particular,
NEURON, is recommended. It is available free to students at the website:
http://neuron.duke.edu/. A version of NEURON which includes a laboratory handbook,
Neurons in Action: Computer Simulations with NeuroLab, by John W. Moore and Ann E.
Stuart, can be purchased from Sinauer Press.

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OPERATION OF METANEURON
MetaNeuron simulations run automatically when the program is opened. A lesson
is selected by clicking the appropriate tab at the top of the screen. The five lessons in
MetaNeuron run independent of each other. If the parameters in one lesson are changed,
the changes will not affect the operation of the other lessons.

Changing parameter values

Experiments are run in MetaNeuron by changing the values of the parameters
displayed in the top portion of the screen. Parameter values can be changed in several
ways:
1) Click on the up or down arrow buttons to the right of a parameter box to increment
or decrement the value by 1. Pressing the "Shift" key while clicking a button increments
the value by 10. Pressing the "Ctrl" key while clicking a button increments values by 0.1.
When running MetaNeuron on an older computer, the traces may update slowly.
Continuous updates can be disabled by unchecking the “Continual Update” box.
2) Click on a parameter box. Keeping the mouse button pressed, drag the mouse.
Moving the mouse up or to the right increases the value by 1. Moving the mouse down
or to the left decreases the value. As in method “1”, the “Shift” and Ctrl” keys change the
increments to 10 and 0.1, respectively.
3) Click on the number in a parameter box, type in a new value, hit "Enter".

MACINTOSH VERSION: The “Shift” and Ctrl” key options and dragging the mouse
(method 2) do not work on Macintosh computers.

Parameters in boxes that are grayed out cannot be changed. The values of these
parameters are determined by other parameters on the screen or are not currently
activated. In the Axon Action Potential and Axon Voltage Clamp lessons, parameters for
Stimulus 2 are activated when the Stimulus 2 On box is checked.
All parameters in a lesson can be reset to their default values by clicking the Reset
to Default button.

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Graphs of parameter values
Some of the parameters in a lesson are plotted in the graph in the lower portion of
the screen. The plots are color coded and corresponding plot labels are shown in the
upper left corner of the graph. Traces in the graph are updated every time a parameter
value is changed. When a parameter value is changed by moving the mouse (PC version
only), the traces update continually. When running MetaNeuron on an older computer,
this continual update may cause annoying delays. If this is the case, continual updates
can be disabled by unchecking the “Continual Update” box. Traces will then be updated
when the left mouse button is released after changing a parameter value.
The sweep duration, the total time displayed on the X-axis of the plots, is
controlled by the Sweep Duration parameter.

Family of curves
A family of curves can be generated in a graph by retaining previous traces. This
is done by checking the “Keep Previous Graphs” box. New traces are added to the graph
every time a parameter value is updated. All traces are retained as long as the box
remains checked. An easy way to generate a family of curves is to check the “Keep
Previous Graphs” box, hold down the Shift key (to increment in steps of 10), left click on
a parameter box (e.g. stimulus amplitude) and move the mouse to the right while keeping
the left mouse button pressed. Examples are shown in figures 5, 7 and 8.

Measuring traces with the cursor

The X and Y values of any point on a graph can be determined by moving the
mouse over the graph and clicking the left mouse button. The X and Y values, in units
appropriate for the yellow trace, are given in the upper right corner of the program
window. The currently selected point on the graph is indicated by crosshairs when the
“Show Cursor” box is checked. The crosshairs display is not shown when the “Keep
Previous Graphs” feature is selected.
When measuring the value of a point on a trace, it is convenient to confine the
cursor to the trace. This is done by checking the “Trace” box. With the box checked, the
Y values of the cursor are confined to points along the yellow trace, which represents

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membrane potential in some lessons and membrane current in others. To measure the
peak voltage of an action potential in the Axon Action Potential lesson, for instance, first
check the “Show Cursor” and “Trace” boxes. Then move the mouse to the X position
corresponding to the peak of the action potential while keeping the left mouse button
pressed.
MetaNeuron represents 1 cm2 of neuronal membrane. The membrane has a
capacitance of 1 µF.

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LESSON 1, RESTING MEMBRANE POTENTIAL
Lesson 1 illustrates how K+ and Na+ channels contribute to the generation of the
resting membrane potential in neurons. The neuron in this lesson is modeled by passive
conductances to K+ and Na+. These conductances are voltage-independent and the
neuron does not generate action potentials.
The concentrations of K+ and Na+, both outside and inside the cell, can be varied.
The program calculates the electrochemical equilibrium potential for each ion, based on
the ion concentration gradient across the membrane, using the Nernst equation,

 [ion]o 
ion equilibrium potential (mV) = 58 ⋅ log 
 [ion ]i 

where [ion]o and [ion]i are the concentrations of the ion outside and inside the cell,
respectively. The effect of temperature on the equilibrium potential is not included in
this version of the equation.
The resting membrane potential of the neuron is determined by the concentrations
of K+ and Na+ outside and inside the cell and by the permeability of the membrane to K+
and Na+. The relative membrane permeabilities to K+ and Na+ can be varied. The
membrane potential is calculated using the Goldman-Hodgkin-Katz equation,

 PK [ K + ]o + PNa [ Na + ]o 
membrane potential (mV ) = 58 ⋅ log + +


 PK [ K ]i + PNa [ Na ]i 

where PK and PNa are the relative membrane permeabilities to K+ and Na+, respectively.
The term representing membrane permeability to Cl- is omitted in the MetaNeuron
simulation. Active membrane conductances are not modeled in this lesson.

Student exercises
1) Vary the concentrations of K+ and Na+, both inside and outside the cell. What
effect does this have on the electrochemical equilibrium potential of the ion? What is the

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Na+ equilibrium potential when Na+ out equals 100 mM and Na+ in equals 10 mM?
What is the Na+ equilibrium potential when Na+ out equals 100 mM and Na+ in equals
100 mM? What is the K+ equilibrium potential when K+ out equals 10 mM and K+ in
equals 100 mM? Why is this?

2) Starting with the default parameter values, vary the relative membrane permeability
to K+ and Na+. What effect does this have on the resting membrane potential? Set Na+
out to 100 mM, Na+ in to 10 mM, K+ out to 10 mM and K+ in to 100 mM. What is the
membrane potential when the Na+ permeability equals 0 and the K+ permeability equals
10? What is the membrane potential when the Na+ permeability equals 10 and the K+
permeability equals 0? What is the membrane potential when the Na+ permeability
equals 1 and the K+ permeability equals 10? Why is this?

3) Starting with the default parameter values, plot the value of the membrane potential
as a function of [K+]o over a [K+]o range of 0.2 to 100 mM. Regraph the data with the
membrane potential plotted as a function of log([K+]o). Can you explain why this second
plot has the shape that it does?

4) Reduce PNa to 0 and replot the membrane potential as a function of log([K+]o). What
accounts for the difference in the shape of this relation, compared to the one in
exercise 3?

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LESSON 2, MEMBRANE TIME CONSTANT
Lesson 2 illustrates the effect of the membrane time constant, τ, on the time
course of electrical responses in neurons. The neuron in this lesson is modeled by a
passive membrane resistance, R, a membrane capacitance, C, and a current source. The
membrane capacitance equals 1 µF/cm2, or simply 1 µF, since MetaNeuron represents 1
cm2 of membrane. The value of the membrane resistance as well as the time course and
amplitude of the current source can be varied. The neuron does not generate action
potentials.
The membrane resistance and capacitance together determine the time constant of
the membrane, as described by the equation,

τ = R ⋅C

The default values of the stimulus current source specify a positive, 1 msec long
pulse. This is a good approximation of the current generated when a fast excitatory
synapse is activated. Using the default current source parameters, MetaNeuron can be
used to assess the effect of the membrane time constant on the time course of single and
multiple synaptic responses.
The "Threshold" potential shown in the graph indicates the approximate voltage
at which action potentials will be initiated in a neuron. Note, however, that the neuron
model used in this lesson does not include active membrane conductances and will not
generate action potentials when the membrane potential exceeds the threshold value.

Measuring time constants

When a constant current is injected into a neuron, the membrane potential
depolarizes with an exponential time course (assuming that there are no active membrane
conductances). Similarly, when the stimulus is turned off, the membrane potential
returns to the resting membrane potential with an exponential time course.
The rate at which the membrane potential increases and decreases is described by
the time constant, τ, of the exponential. τ is defined as the time it takes for the membrane
to increase or to decrease to 1-1/e (approximately 63%) of its final value. Remember that

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sufficient time must be allowed for the membrane potential to reach its plateau level in
order to make this measurement.

Student exercises
1) Using the default parameter values, determine the time constant of the membrane by
measuring the time it takes for the amplitude of the membrane potential to fall 63% of the
way back to the baseline value, after the current source is turned off. Does this value of τ
equal the membrane time constant calculated from the equation,
τ = R ⋅C
where τ is the membrane time constant in units of sec, R is the membrane resistance in
units of Ohms·cm2 and C is the membrane capacitance in units of F/cm2?
Increase the amplitude of the stimulus to 150 µA and repeat your measurement of
the membrane time constant. Has it changed?

2) Using a stimulus amplitude of 150 µA, vary the membrane resistance. What effect
does this have on the rising and falling phases of the response? Set the membrane
resistance to 2 k Ohms·cm2. What is the membrane time constant, measured from the
falling phase of the response?

Figure 1. Membrane time constant.

A 150 µA, 1 msec current pulse
rapidly depolarizes a neuron. At
the end of the pulse, the membrane
potential decays back towards the
10 k Ohms resting membrane potential with an
exponential time course. The time
2 k Ohms
constant of decay is more rapid for
a membrane resistance of 2 k Ohms
than for a resistance of 10 k Ohms.
stimulus

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3) Reset to the default parameter values and increase the number of stimuli from 1 to 3.
Note how the responses add together. This superposition of responses is termed temporal
summation. Now reduce the membrane resistance from 10 to 2 k Ohms·cm2. What
effect does this have on temporal summation? Note that changes in temporal summation
determine whether the response reaches the threshold level for firing action potentials.

Figure 2. Temporal summation.

The three current pulses (bottom
trace) simulate three synaptic
responses. With a long membrane
time constant (10 k Ohm
membrane resistance), the synaptic
potentials summate and reach the
threshold potential for firing an
"threshold" action potential. With a short time
membrane potential constant (2 k Ohms), the synaptic
potentials do not summate and do
not reach threshold.
stimulus

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LESSON 3, AXON ACTION POTENTIAL
Lesson 3 illustrates how voltage- and time-dependent Na+ and K+ conductances
generate the action potential. The MetaNeuron model in this lesson is based on equations
developed by Alan Hodgkin and Andrew Huxley to describe the voltage-dependent Na+
and K+ conductances of the squid giant axon. The Na+ conductance in the MetaNeuron
model represents fast, tetrodotoxin-sensitive Na+ channels. The K+ conductance
represents delayed rectifier K+ channels. The values of some of the parameters in the
equations have been adjusted to simulate action potential generation in a vertebrate axon.

Several parameters controlling Na+ and K+ currents can be adjusted in the model:
• The Na+ and K+ equilibrium potentials. Modifying these values will change the
driving force for ion flux through the Na+ and K+ conductances.
• gNa+ max and gK+ max, the maximum membrane conductances. These
parameters represent the number of Na+ and K+ channels in the membrane.
• TTX (tetrodotoxin) and TEA (tetraethylammonium). Selecting TTX blocks the
Na+ conductance. Selecting TEA blocks the K+ conductance.
• Temperature. Raising the temperature reduces the time constants controlling Na+
conductance activation and inactivation and K+ conductance activation, resulting
in a faster action potential. The temperature factor has a Q10 of 3.0.

Activation of the neuron is controlled by several stimuli:

• Stimulus 1. The delay, width and amplitude of this current pulse can be varied.
• Stimulus 2. A second current pulse is added when the Stimulus 2 On box is
checked. The second stimulus is used when measuring the refractory period of
the axon.
• Holding Current. This continuous current source can be used alone or together
with the current pulses.

The graph displays the membrane potential as well as the Na+ and K+ equilibrium
potentials and the stimulus. In addition, Na+ and K+ conductances or currents are

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displayed when the appropriate buttons are selected in the Conductances and Currents
box.

Student exercises
1) Increase the amplitude of Stimulus 1. What effect does this have on the cell
response? What is the threshold stimulus amplitude (in µA) and the threshold voltage (in
mV) for initiating an action potential? (The threshold voltage is best measured by
adjusting the stimulus amplitude, in 0.1 µA increments, until it is just below threshold for
initiating an action potential. Now measure the voltage at the peak of the response.
Measure the voltage using the cursor, with the Show Cursor and Trace boxes checked.)

Figure 3. Action potential

Na+ potential
generation. Injection of current
into an axon triggers the generation
of an action potential. The peak of
the action potential approaches the
0 mV
Na+ equilibrium potential. The
after hyperpolarization reaches the
membrane potential
K+ equilibrium potential.

K+ potential
stimulus

2) Vary the stimulus amplitude from 50 to 200 µA in 10 µA increments. Measure the

amplitude of the action potential and the time to the peak of the action potential for each
stimulus amplitude. Plot both the amplitude and time to peak of the action potential as a
function of stimulus amplitude. Why does the time to peak vary? Why does the action
potential amplitude vary so little?

3) Action potentials are often initiated at the axon hillock, where the axon leaves the
cell body. The axon hillock has a high density of Na+ channels, which lowers the
threshold for initiating an action potential. MetaNeuron can be used to measure the effect
of Na+ channel density on the threshold for action potential initiation. Vary gNa max

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from 200 to 380 mS/cm2 in 20 mS/cm2 steps. (Na+ channel density is proportional to
gNa max.) For each gNa max value, determine the threshold for initiating an action
potential. Measure both the threshold stimulus amplitude (in µA) and the threshold
voltage. Plot the threshold current and voltage as a function of gNa max. Why does the
action potential threshold vary as Na+ channel density is changed?

4) The symptoms of multiple sclerosis are caused by conduction failure due to the
demyelination of axons. In some patients, the symptoms of multiple sclerosis are
alleviated by cooling a patient while symptoms are exacerbated by an exercise-induced
rise in body temperature. MetaNeuron can be used to measure the effect of temperature
on action potential threshold. Adjust the stimulus amplitude so that it is just below
threshold for initiating an action potential. Now vary the temperature. How much does
the temperature have to be lowered to evoke an action potential? Starting again from the
default parameter values, adjust the stimulus amplitude so that it is just above threshold
for initiating an action potential. How much does the temperature have to be raised to
block initiation of the action potential? Why does action potential threshold depend on
temperature?

5) Set Stimulus 1 Amplitude to 150 µA. The stimulus will evoke an action potential.
Vary the Na+ potential. What effect does this have on the action potential? Why?

6) Using a Stimulus 1 Amplitude of 150 µA and a Na+ potential of 50 mV, vary gK

max. What effect does this have on the width of the action potential? Why?

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 7) Starting with the default parameter values, set Stimulus 1 Amplitude to 0 and Sweep
Duration to 20 ms. Now raise the K+ potential (in the Membrane Parameters box) to
more positive values. As the K+ potential is raised, you will observe the following
sequence of events,
• The membrane potential depolarizes.
• Spontaneous action potentials are generated.
• The frequency of the action potentials increases.
• The amplitude of the action potentials diminishes.
• The action potentials die out as the membrane potential continues to depolarize.
Can you explain this sequence of behavior?

8) Starting with the default parameter values, set Sweep Duration (the total time
displayed on the X-axis) to 10 msec. Now decrease the amplitude of Stimulus 1 until it
reaches –200 µA. You will observe that an action potential is evoked by the
hyperpolarizing stimulus! This phenomenon is called anodal break excitation. Can you
explain why a hyperpolarizing stimulus evokes an action potential?

9) MetaNeuron can be used to measure the time course of the refractory period which
follows an action potential. Two stimulus pulses are used to measure the refractory
period. The amplitude of the first stimulus is set to well above threshold and reliably
evokes an action potential. The amplitude of this stimulus remains fixed. The second
stimulus, which is initially set to a delay of 12 msec after the first stimulus, is used to
determine the change in threshold of the axon. The amplitude of the second stimulus is
adjusted until it is just strong enough to evoke a second action potential. The stimulus
amplitude is a measure of the threshold of the axon. The higher the threshold of the
axon, the stronger is the stimulus needed to trigger the second action potential.
Starting with the default parameter values, set Sweep Duration to 16 msec and
Stimulus 1 Amplitude to 150 µA. Turn Stimulus 2 ON and set Stimulus 2 delay to 12
msec.

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 With Stimulus 2 Delay at 12 msec, adjust Stimulus 2 Amplitude until it is just
large enough to evoke an action potential. This is the threshold amplitude. (The threshold
amplitude can be easily determined on PC computers by clicking on the Stimulus 2
Amplitude box and dragging the mouse while keeping the left mouse button pressed.
Move the mouse until an action potential is evoked.)
Reduce Stimulus 2 delay to 11 msec and once again adjust Stimulus 2 Amplitude
to the threshold level. Repeat this measurement for Stimulus 2 Delays of 10 to 1 msec.
As you approach a Stimulus 2 Delay of 1 msec, reduce the delay in smaller steps so that
the threshold amplitude vs. delay relation can be determined more accurately. Note that
at small delays, the second action potential becomes smaller and is not an “all or none”
event. Use your judgment in determining a criterion for when the action potential is
evoked.
Plot the Stimulus 2 threshold amplitude as a function of Stimulus 2 delay. The
plot shows the time course of the refractory period following an action potential. What is
the duration of the absolute refractory period and the relative refractory period, based on
this experiment?

Figure 4. The duration of the

refractory period following an
action potential can be measured
using a two pulse protocol. The
first stimulus pulse is well above
threshold and reliably evokes an
action potential. The second
stimulus pulse is adjusted so that
it just triggers an action potential.
The amplitude of the second
stimulus is a measure of the
membrane threshold.

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LESSON 4, AXON VOLTAGE CLAMP

Lesson 4 uses the voltage clamp technique to reveal the properties of the Na+
channels and delayed rectifier K+ channels that generate the action potential. Na+ and K+
channels are both voltage- and time-dependent and it is difficult to analyze their
properties while action potentials are being generated. Channel properties can be
characterized, however, by measuring the currents flowing through the channels at a
fixed membrane potential. The voltage clamp technique, developed by Hodgkin and
Huxley in the 1940’s, permits these measurements to be made.
The voltage clamp technique uses a negative feedback electronic circuit to hold
the axon membrane potential to a value (the command voltage) specified by the
experimenter. The circuit then measures the time-dependent currents flowing through
Na+ and K+ channels at that voltage. Typically, the membrane potential is first held at a
negative voltage near the resting membrane potential, the Holding Potential, and is then
stepped to a more positive voltage, determined by the Stimulus 1 and Stimulus 2
Amplitudes in MetaNeuron.
It is important to remember that action potentials cannot be generated when an
axon is voltage clamped. The currents that are measured under voltage clamp contribute
to the generation of the action potential, but these currents are not, in themselves, action
potentials.
Lesson 4 uses the same Hodgkin-Huxley model of the axon that is used to
generate action potentials in Lesson 3. The parameters in Lesson 4 are similar to those in
Lesson 3 as well. The only changes in Lesson 4 are that the axon is now voltage-clamped
and the stimuli used are command voltages rather than currents. The Holding Potential
parameter determines the voltage of the axon at the beginning of the experiment. The
Stimulus 1 and Stimulus 2 Amplitudes determine the voltage to which the membrane is
stepped during the two pulses. Note that these stimulus amplitudes represent absolute
voltages, not relative voltages added onto the holding potential.

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 Student exercises
1) Increase Stimulus 1 Amplitude and note the development of a transient inward
(negative) current and a sustained outward (positive) current. What is the origin of these
two components of the current?

membrane current
Figure 5. Transient inward Na+
currents and sustained outward K+
currents are recorded when the
axon is voltage clamped. This
family of eight voltage clamp
recordings was generated by
stepping the membrane potential
from -70 mV to +70 mV in 20 mV
increments.
command voltage

2) Starting with the default parameters, set the Stimulus 1 Amplitude to 0 mV. What
happens when the TTX or TEA boxes are selected? Why is this?

3) The K+ current can be viewed in isolation by blocking the Na+ current with TTX.
With the TTX box checked, measure the amplitude of the K+ current as a function of the
voltage of the test pulse (Stimulus 1 Amplitude). The K+ current should be measured
from its pre-pulse baseline level (which you can assume equals 0) to its plateau level, at
the end of the voltage pulse. Measure the current over a stimulus amplitude range of –70
to +70 mV in increments of 10 mV. Plot the amplitude of the K+ current as a function of
the test pulse voltage.
The K+ current can be converted to a conductance using the chord conductance
equation,
current
conductance =
Vm − Vequilibrium

where Vm is the test pulse voltage and Vequilibrium is the equilibrium potential for the ion,

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which does not change between trials. Use this equation to calculate the K+ conductance
at each test pulse voltage and plot the conductance as a function of voltage. What does
the conductance-voltage relation tell you about the voltage dependence of K+ channels?

4) Repeat this procedure to determine the current-voltage and conductance-voltage

relations for Na+. Select the TEA box and de-select the TTX box in order to view the
Na+ current in isolation. Measure the amplitude of the Na+ current as a function of
voltage over a stimulus amplitude range of –70 to +80 mV. Measure the Na+ current
from the pre-pulse level to the transient peak of the current. Note that the peak current
will be negative at some voltages and positive at other voltages. Plot the current-voltage
relation. Convert the currents to conductances using the chord conductance equation and
plot the conductance-voltage relation. What does the conductance-voltage relation tell
you about the voltage dependence of Na+ channels? In the current-voltage relation, why
is Na+ current negative at some voltages and positive at other voltages?

5) Na+ channels first activate and then inactivate when the membrane potential is
stepped from the resting level to a depolarized level. The channels do not begin to
recover from inactivation until the membrane is repolarized to a negative level.
MetaNeuron can be used to measure the time course of recovery of the Na+ conductance
from inactivation by voltage clamping the axon and using two voltage command pulses.
Starting with the default parameter values, check the TEA box to view the Na+
current in isolation, turn on Stimulus 2, set Sweep Duration to 16 msec, Stimulus 1 and
Stimulus 2 Amplitude to +5 mV, Stimulus 1 and Stimulus 2 Width to 2 msec and Stimulus
2 Delay to 10 msec.
With Stimulus 2 Delay set at 10 msec, measure the amplitude of the peak Na+
current evoked by the second pulse (the current will be negative). Decrease the Stimulus
2 Delay to 9 msec and repeat the measurement. Continue measuring the Na+ current
amplitude for delays down to 0.1 msec, decreasing the delay in 1 msec increments. As
you approach a delay of 0.1 msec use smaller decrements in order to accurately
determine the time course of recovery from inactivation.

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 Plot the amplitude of the second pulse Na+ current as a function of the delay.
How does the time course of recovery from Na+ channel inactivation compare to the time
course of the refractory period following an action potential? Why are these two
processes related?

membrane
current
command
voltage

Figure 6. Recovery from Na+ channel
inactivation can be measured with the
voltage clamp technique using a two pulse
protocol. Stepping the membrane potential
from -75 to +5 mV evokes an inward Na+
current. Following a delay of 1 msec, a
second depolarization evokes a smaller
current, demonstrating that many Na+
channels have not yet recovered from
inactivation. TEA has been selected to
block K+ currents.
Figure 7. The time course of recovery of
Na+ channels from inactivation is revealed
in this family of voltage clamp recordings.
The two pulse experiment illustrated in Fig.
6 was repeated for delays ranging from 0.1
to 7.5 msec. The peaks of the inward Na+
current plots outline the time course of
recovery from inactivation.

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LESSON 5, SYNAPTIC POTENTIAL AND CURRENT
Lesson 5 illustrates the synaptic voltages and currents generated at fast ionotropic
synapses. The neuron in Lesson 5 is modeled by a leak conductance and by the opening
of an ionotropic receptor conductance, simulating a synaptic response. The relative
permeabilities of the receptor to Na+, K+ and Cl- can be adjusted. Based on these relative
permeabilities, the model calculates the reversal potential of the receptor. It is important
to bear in mind that ionotropic receptors are normally permeably to either cations (Na+,
K+) or anions (Cl-) but not to both. The neuron in lesson 5 does not generate action
potentials.
The reversal potential of the ionotropic receptor can be determined
experimentally by varying the membrane potential of the neuron and noting the potential
at which the synaptic response reverses polarity. When synaptic voltages are measured
(Mode: Voltage) the membrane potential is varied by injecting current into the cell. This
is done by adjusting the Holding Current. (This mode of recording is also referred to as
“current clamp”.) When synaptic currents are measured (Mode: Voltage Clamp), the
membrane potential is varied by adjusting the Holding Potential.
The equilibrium potentials for Na+, K+ and Cl- in this model are +50, -77 and -75
mV, respectively. The resting membrane potential is determined by a leak conductance
having a reversal potential of -65 mV. The synaptic conductance is modeled with rising
and falling time constants of 0.1 and 1.0 msec.

Student exercises
1) Start with the default parameter values, with relative receptor permeabilities of Na+,
1.2; K+, 1; Cl-, 0. These default permeabilities represent an excitatory receptor such as
the AMPA glutamate receptor. Depolarize the neuron incrementally by increasing the
Holding Current from 0 to 240 µA in 30 µA increments. What happens to the synaptic
potential, which is shown in the yellow (Membrane Voltage) trace, as the membrane is
depolarized? Plot the amplitude of the synaptic potential as a function of membrane
voltage. Measure the amplitude from the baseline to the peak of the synaptic response.
Note that the amplitude will be negative at some membrane voltages. What is the shape

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of the relation?
The reversal potential of the synapse is defined as the potential at which the
synaptic response reverses polarity. At what voltage does the synaptic potential reverse?
Does this agree with the reversal potential calculated by the model and displayed in the
Receptor Reversal Potential box?

Figure 8. Reversal of an excitatory

postsynaptic potential generated by
Na+ potential
the opening of a receptor permeable
to both Na+ and K+. This family of
voltage responses was made by
depolarizing the neuron by injection
0 mV
of a holding current. The synaptic
response reverses polarity when the
neuron is depolarized past 0 mV.
membrane potential
K+ potential

2) Starting with the default parameters, switch to Voltage Clamp mode. Now
depolarize the membrane incrementally by raising the Holding Potential from -65 to +65
mV in 10 mV increments. Plot the amplitude of the synaptic current as a function of
holding potential. Measure the current amplitude from the baseline to the peak of the
synaptic response. At what voltage does the synaptic current reverse polarity? Is this the
same voltage as was calculated in exercise 1? What is the shape of the synaptic current
vs. voltage relation? Why does the relation have this shape?
The synaptic current can be converted to conductance using the chord
conductance equation,
current
conductance =
Vm − Vreversal
where Vm is the membrane potential specified by the Holding Potential and Vreversal is the
reversal potential of the synaptic response. Use this equation to calculate the synaptic
conductance at each voltage and plot the conductance as a function of voltage. What is
the shape of the conductance vs. voltage relation? Is the synaptic conductance voltage-
dependent?

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 3) Change the relative permeabilities of the receptor to Na+, 0.3; K+, 1; Cl-, 0. In
Voltage Clamp mode, measure the reversal potential of the current. At what voltage does
the current reverse polarity? Why has the reversal potential of the receptor changed?

4) Change the relative permeabilities of the receptor to Na+, 0; K+, 0; Cl-, 1. This
represents an inhibitory receptor, such as the GABAA receptor. In Voltage Clamp mode,
measure the reversal potential of the current. At what voltage does the current reverse
polarity? Why is this synapse inhibitory?

5) Starting with the default parameters, note the time course of the synaptic potential
and measure the time to the peak of the response. With the “Keep Previous Graphs” box
selected, switch to Voltage Clamp mode. Note that the time course of the synaptic
response has changed. Why is this?

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