Neur 3101 / 6101

Cellular Neuroscience
Pre-Lab Tutorial: Simulation 2

John Bekkers
John Curtin School of Medical Research
Australian National University
John.Bekkers@anu.edu.au

Aims of Prac 2

Part 1: Action potential threshold What is the minimum stimulus for an AP?
Part 2: Refractoriness How soon can a second AP be elicited?
Part 3: AP frequency vs stimulus current How fast can APs fire in trains?
Part 4: Voltage clamp Properties of Na and K currents underlying APs?

1!
 Aims of Prac 2

Part 1: Action potential threshold What is the minimum stimulus for an AP?

Vary the amplitude & duration of

the stimulus current step

Measure the current that just

evokes an AP

How does threshold amplitude

depend on duration?
Why?

NOTE: AP threshold is often called the rheobase

Aims of Prac 2

Part 2: Refractoriness How soon can a second AP be elicited?

Vary the amplitude & delay of

the 2nd stimulus current

Determine the conditions for

eliciting a second AP

Relative vs absolute
refractory interval
Why this behaviour?

NOTE: An AP is defined to occur when its peak is above 0 mV

2!
 Aims of Prac 2

Part 3: AP frequency vs stimulus current How fast can APs fire in trains?

Vary the amplitude of the stimulus

current (a 1 s-long step)

Count the number of APs that

are elicited frequency of APs

Plot an F-I curve

What is the shape of this curve

and why?

NOTE: The program automatically counts the number / frequency of APs

Aims of Prac 2

Part 4: Voltage clamp Properties of Na and K currents underlying APs?

Impose (clamp) a Vm step

Measure the peak INa & IK as well

as gNa and gK
gNa!
Normalised

Calculate peak conductance of INa & IK

using I = g (Vm Eion) to find g
Measure steady-state inactivation of INa -120 -80 -40 0 +40
Membrane potential (mV)!

3!
 Running the simulations

1. Double-click on NEUR3101_II and proceed as last time

2. Read the hints at the beginning of the lab notes: baseline

Measure all amplitudes as a difference between two cursors,
one of them placed on the baseline
Move cursor with left / right arrow keys
Magnify areas using the mouse to drag a box around the area
of interest, click in the box, then select Expand
Revert to original axes by Edit Undo Scale Change
peak
Alternative: Double click on axis and enter range manually

3. Strategy for finding thresholds etc:

Find the right ballpark value by making coarse guesses, then
home in by making small changes

Passive membrane properties

Action potential threshold: Refractoriness:

Action potential
(active response) Action potential

Passive or subthreshold Passive response

response

4!
 Passive membrane properties

Istim The electrical resistance between the

Voltage-independent inside and outside of the cell via all the
leak channels
(determine Erest)
Vm leak channels is R (voltage-independent)

Istim R is commonly called:

RN = resistance of the neuron, or
Rin = input resistance of the neuron

If we inject a current Istim into the cell, the

voltage across the membrane, Vm, will
change by an amount given by Ohms Law:
V = Istim Rin

Vm: V

Istim:

Passive membrane properties

Istim However, the cell membrane has a

Voltage-independent capacitance (also voltage-independent)
leak channels
(determine Erest)
Vm called Cm

Istim Cm stores charge (remember the rubber

balloon analogy) and slows down
changes in Vm
Hence, the change in Vm is slowed down at
the start and end of the Istim step:
Cm The change in Vm follows an exponential
decay, with a time constant called the
membrane time constant, m (tau-m)

Vm:

Istim:

5!
 Passive membrane properties

Istim This passive response scales up and

Voltage-independent down as Istim is made larger and smaller,
leak channels
(determine Erest)
Vm but it does not change its shape

Istim The passive response is due to the

intrinsic properties of the membrane,
and is always present when a
stimulus current is applied

Cm

Vm:

Istim:

Passive membrane properties

Istim Activation of voltage-dependent ion

Voltage-independent channels (e.g. NaV or KV channels) now
leak channels
(determine Erest)
Vm adds active membrane responses

Istim The active responses sit on top of the

passive responses
Active response

Passive response
Cm

Vm:

Istim:

6!
 Passive membrane properties

Istim The input resistance, Rin, of the cell can be

Voltage-independent calculated from the plateau value of Vm
leak channels
(determine Erest)
Vm using Ohms Law

Istim However, if Istim lasts for much less time

than the membrane time constant, the
Active response plateau will not be reached

Passive response

Cm V
Rin =
Istim

Vm: V

Istim:

Hints for writeup (see also Lab Handbook 2017)

Paraphrase the aims of each part of the experiment

Present primary data (tables, plots) but be selective in what primary plots you show

Plots:
Dont simply connect the dots

If you believe the data should fit a straight line,

fit the line by eye or by computer and give
the fit parameters (slope, intercept)

If you dont think the data fit a straight line, you

can superimpose a curve (by eye) or a spline
or polynomial fit (by computer), but there is
no need to give the fit parameters

See the Lab Handbook for other advice about

labelling axes, statistics, etc.

Answer the questions and do the things in the boxes in the lab notes

Be succinct!
Reports are due 5 p.m. Mon 20 Mar

7!