ECE 795:

Quantitative
Electrophysiology
Notes for Lecture #2
Wednesday, September 20, 2006
3. CELL EXCITABILITY
Here we will look at the response of the cell
membrane to current injections and changes
in the transmembrane potential.
The key features are:
1. the linear behaviour around the resting
membrane potential, and
2. the nonlinear behaviour that leads to the
generation of “action potentials”.
2
Equivalent circuit near rest:

(from Koch)
3
Notation for potentials:
¾ Transmembrane potential:

¾ Membrane potential relative to rest:

¾ Intra- and extra-cellular potentials relative to

their respective baseline values:

4
Spherical cell response to current step:
If Rm (Ω·cm2) and Cm (μF/cm2) are the specific
resistance and the specific capacitance,
respectively, of the membrane, then for a spherical
cell with surface area A (cm2) the membrane
resistance is:

and the membrane capacitance is:

5
Spherical cell response to current step
(cont.):

6
Spherical cell response to current step
(cont.):
The response of the relative transmembrane
potential vm of a spherical cell subjected to an
intracellularly-injected current step of amplitude I0
is:

where τ = RC and V0 = I0R = vm(t→∞).

7
Spherical cell response to current step
(cont.):
I0
i(t)

0
τ = 10 ms
V0
vm(t)

0
0 20 40 60 80 100
8
t (ms)
Strength-duration relationship:
Suppose that a relative transmembrane potential
of VT (< V0) is the “threshold” potential for eliciting
an action potential.
From Eqn. (7.4), the membrane potential will reach
VT at time T (following the onset of the current
pulse) according to the equation:

Solving for T gives:

9
Strength-duration relationship (cont.):

I0
i(t)

0
τ = 10 ms
V0

VT
vm(t)

0
0 T 20 40 60 80 100
t (ms) 10
Strength-duration relationship (cont.):
A strength-duration curve is obtained by plotting:

for a fixed value of VT.

11
Strength-duration relationship (cont.):
¾ Rheobase is the minimum excitation required
to just reach threshold as T → ∞, i.e., V0 = VT.
¾ Chronaxie is the pulse duration Tc required to
reach threshold when the stimulus is twice
rheobase, which can be calculated according
to:

Chronaxie is significant as a nominal time

period required to reach the threshold voltage.

12
Action potentials are:
¾ all-or-nothing events,
¾ regenerative,
¾ generated when a threshold is reached,
¾ propagating potentials, and
¾ also known as nerve spikes or impulses.

13
Transmembrane action potential
morphology:

14
Observing
action
potentials
(cont.):

(from Koch)

15
Nonlinear membrane behaviour:
Subthreshold and
action potential
responses to a
brief stimulating
current.

16
Nonlinear membrane behaviour (cont.):

17
Nonlinear membrane behaviour (cont.):
For the squid axon:-
¾ Cm ≈ 1 μF/cm2 throughout the entire
action potential
¾ Rm ≈ 1000 Ω·cm2 at rest
¾ Rm ≈ 25 Ω·cm2 at the peak of the action
potential

18
Origin of action potential, resting and
peak voltages:
In the classic studies by Hodgkin and
Huxley, the results were related to the
Goldman-Hodgkin-Katz (GHK) equation for
the transmembrane potential:

where Pp is the permeability of the pth ion

channel.
19
Origin of action potential, resting and peak
voltages (cont.):
As we have observed previously, the
resting transmembrane potential is
slightly higher than the potassium
equilibrium potential.
Looking at the action potential waveform, we
see that the peak transmembrane
potential approaches but never exceeds the
sodium equilibrium potential.

20
Origin of action potential, resting and peak
voltages (cont.):
This result is consistent with an elevated sodium
permeability in the rising phase and peak of the
action potential.
Good agreement between theory and experimental
data from the squid axon is obtained with:

21
Origin of action potential, resting and peak
voltages (cont.):
To a first approximation:

22
Origin of action potential, resting and peak
voltages (cont.):
In an experiment using radioactive tracers, it
was found for the cuttlefish Sepia giant axon
that:
¾ at rest, there is steady influx of sodium and
efflux of potassium, consistent with
EK < Vrest < ENa
¾ during an action potential there is an influx of
3.7 pmoles/cm2 of sodium
¾ during an action potential there is an efflux of
4.3 pmoles/cm2 of potassium
23
Origin of action potential, resting and peak
voltages (cont.):
These results can be compared with the
charge movement required to depolarize the
transmembrane potential by around 125 mV:

24
4. INTRODUCTION TO THE HODGKIN-
HUXLEY MODEL

Based on a set of detailed physiological

experiments, Hodgkin & Huxley developed a
nonlinear dynamical model of the excitable
cell membrane. Their formulation for gating
dynamics is the basis of almost all
subsequent biophysical models of excitable
cell membranes.

25
Voltage and space clamp:
¾ Hodgkin and Huxley used a voltage and space
clamp apparatus to measure and quantify ionic
currents in the squid giant axon.
¾ By applying a voltage clamp and making
discrete steps in transmembrane voltage, the
capacitive current is absent, and consequently
only the ionic currents are recorded.
¾ By inserting a conducting wire along the inside
of the axon, a space clamp is applied, i.e., the
intracellular potential is the same along the
entire length of the axon. Consequently, the
ionic current from a large number of ion
channels is recorded. 26
Voltage and space clamp (cont.):

27
Example net ionic current to a voltage step:

28
Measured ionic current for different voltage
steps:

29
Current-voltage curves:

30
Separation of sodium and potassium
currents:
Hodgkin and Huxley utilized two approaches
to separating the contributions of sodium
and potassium currents to the net ionic
current:
1. Assuming IK = 0 for 0 · t · T/3, where
T is the time of peak inward current.
2. Varying the extracellular sodium
concentration while keeping the
extracellular potassium concentration
fixed. 31
Effects of varying extracellular sodium
concentration:

32
Ionic conductances from ionic currents:
Rearranging Eqns. (3.26) and (3.27) gives:

In the voltage clamp experiments, the

denominators of Eqns. (5.11) and (5.12) are
constant ⇒

33
Ionic conductances from ionic currents
(cont.):

34
Hodgkin-Huxley equations:
Potassium channel model:
¾ Potassium conductance:

¾ Potassium activation particle dynamics:

or:

35
Hodgkin-Huxley equations (cont.):
¾ Potassium activation transition rates:

36
Hodgkin-Huxley equations:
Sodium channel model:
¾ Sodium conductance:

¾ Sodium activation particle dynamics:

¾ Sodium inactivation particle dynamics:

37
Hodgkin-Huxley equations (cont.):
¾ Sodium activation transition rates:

¾ Sodium inactivation transition rates:

38
Hodgkin-Huxley equations (cont.):

39
Hodgkin-Huxley equations (cont.):

(from Koch) 40
Simulation of membrane action potential:
The complete Hodgkin-Huxley model is a parallel-
conductance model incorporating:
¾ nonlinear (active) potassium and sodium
currents:

¾ a linear (passive) “leakage” current:

¾ and a capacitive current: 41

Simulation of membrane action potential
(cont.):
In the case of the an injected depolarizing current
Id, the membrane equation is:

With the equations describing the gating particle

dynamics, we have one first-order nonlinear
differential equation coupled with three first-order
linear differential equations.
Analytical solutions are not very tractable, so in
most cases it is necessary to find numerical
solutions. 42
Simulation of membrane action potential
(cont.):
¾ Hodgkin and Huxley had assistants doing
numerical solutions by hand – not much fun!
¾ Matlab has a set of numerical ODE solvers.
¾ Software packages for simulating neurons
include:
– HHsim: Graphical Hodgkin-Huxley Simulator
– NEURON: For computer simulations of neurons
and neural networks
– The GEneral NEural SImulation System
(GENESIS)
43
Simulation of membrane action potential
(cont.):

44
Action potential characteristics:
The characteristics of action potentials can
be interpreted in terms of:
¾ how the ionic and capacitive currents
vary as a function of time, membrane
potential and injected current,
The behaviour of the ionic currents is
understood in terms of:
¾ voltage-dependent channel gating, i.e.,
the dynamics of activation and
inactivation particles.
45
Action potential characteristics (cont.):

(from Koch)

46
Threshold behaviour:
Consider the case where the membrane is at rest
and is then depolarized very rapidly to a relative
membrane potential of vm. If sodium activation is
assumed to occur instantaneously and sodium
inactivation and potassium activation are assumed
to remain unchanged, then the net ionic current is:

47
Threshold behaviour (cont.):
The resulting current-voltage relationship, shown
below, explains the threshold behaviour of such a
depolarization.

(from Koch)

48
Accommodation:
Very slow changes in the membrane
potential allow sodium inactivation and
potassium activation to overcome sodium
activation.
Consequently, the cell may not spike, even
though the nominal threshold potential (i.e.,
in the case of a rapid depolarization) has
been reached.
Any definition of a “threshold potential” is
therefore restricted to a particular stimulus.
49
Accommodation (cont.):

50
Accommodation (cont.):

51
Strength-duration behaviour:
For a finite-duration current pulse, the strength of
the stimulating current required to just elicit one
action potential is characterized by a strength-
duration curve.

52
Anode break excitation:
Sodium deinactivation
and potassium
deactivation can give
rise to an action
potential at the offset
of a hyperpolarizing
pulse. This is referred
to as “anode break”
excitation.

53
Repetitive firing:
Injection of a sustained suprathreshold current
gives rise to repetitive firing, illustrating the
regenerative nature of spiking.

(from Koch)

54
Repetitive firing (cont.):
Increasing the injected current produces a slight
increase in the spike rate.

(from Koch)

55
Refractory effects:
¾ During most of the falling phase of an
action potential it is impossible to
generate another action potential,
irrespective of the magnitude of injected
current. This is referred to as the
absolute refractory period.
¾ For some time following an action
potential the injected current required to
reach threshold is greater than is
necessary when the membrane is at rest.
This is referred to as the relative
refractory period.
56
Refractory effects (cont.):

57
Refractory effects (cont.):

58
Refractory effects (cont.):

59
(from Koch)