Neurophysiology – lecture 17

March 15, 2011

1 Review:
1. In the week before vacation we discussed:
(a) Voltage clamping – using an apparatus which holds the transmembrane potential constant while
measuring the current needed to do this.
(b) The total I (IT ) needed to hold the Vm constant flows across the cell membrane as the sum of IK
and INa and IL and Ic .
(c) We were able to subdivide IT into the individual currents IK and INa . In fact entire families
ofIK andINa curves at various transmembrane potentials were defined for the 10 or 20 ms after a
voltage step was applied.
(d) From these families ofIK andINa curves, we could determine gK and gNa curves using the equations:

IK INa
gK = and gNa =
Vm − EK Vm − ENa

(e) From the evaluation of gK and gNa we saw that both of these conductsances increase from a value
near 0 when the Vm is near -60mV to a maximal value when Vm equals approximately +10mV.
(See Figure 11B.)
2. Resynthesizing A.P.s
(a) In a voltage clamp only currents at particular voltages can be measured.
(b) During normal intracellular recording only voltages can be measured, not currents.
(c) Thus there is no way to bring together both current and voltage measurements to describe how
action potentials are produced.
(d) However, the output from voltage clamp recordings can be incorporated mathematically with the
voltages recorded during regular recordings to provide a mathematical model of how A.P.s are
generated.
(e) In this mathematical model: a slight deploarization →
a small increase in gNa and a later increase in gK →
a small increase in INa and a later increase in IK →
Inet being inward →
an increase in intracellular +charge, dq = Inet dt, where dt is a somewhat arbitrary short period
of time

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 dq
an increase in dq produces a further depolarization, dvm where dvm = (And we are back to
Cin
the initial step).
(f) To this cycle of processes we added another step which is that the slight depolarization actually
changes:
m – the Na channel activation parameter – where gNa = m3 h gNamax
n – the K channel activation parameter – where gK = n4 gKmax
h – the Na channel inactivation parameter – where gNa = m3 h gNamax
(g) Important properties of this mathematical model include:
a “positive feedback” loop since a small depolarization can initiate a larger depolarization through
Na channel activation.
It also includes 4 “negative feedback” loops since:
i. maintained depolarization increases gK → more IK
ii. maintained depolarization decreases h → a smaller gNa
iii. depolarization reduces the driving force on Na and hence reducesINa
iv. depolarization increases the driving force on K and hence increases IK

2 Demonstration of the processes involved in the mathematical

model
1. The computer display was then used to show that the sequence of calculations described above was
actually taking place when the computer program created the display of an action potential.
2. To make this possible the program was slowed down by inserting delays at several spots in the program.
3. The numeric value of each calculated gNa and gK was printed out in the upper right hand corner of
the display together with the IK and INa which were calculated and the final Vm . Throughout the
development of the A.P. these numbers were being sequentially calculated and then displayed.

3 Mathematical Model explanation of known A.P. properties

1. The concept of Threshold
(a) In introductory books Threshold is generally described as a membrane depolarization which ex-
ceeded a certain value and produced an action potential.
In class on Thursday (3/3/2011) Sean Moore1 suggested that Threshold would be reached when
there was a net inward ionic current; that is, when inward INa was greater that the outward IK .
2. So using the computer simulation model we showed that a very small change in stimulation could be
sufficient to produce an A.P. where there previously was none. A stimulation of 7.67 for 1 ms did not
produce an A.P., but a stimulation of 7.68 for 1 ms did elicit an A.P.
1 Yeah, Sean!

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 Looking closely at the current traces produced by the computer it was evident that inward INa increased
very slightly well before the sharp change seen in Vm seen at the onset of the action potential, whereas
IK did not change until later.
Thus the definition of Threshold as something that occurs when INet is inward appears to hold.
3. I then asked if gK = gNa what would INet be equal to? As per custom no one answered which in this
case was wise, since the answer could not be easily determined.
Since
gNa gK
INa = and IK = at Vm = −60 mV
Vm − ENa Vm − EK
(Vm − ENa ) is absolutely larger than (Vm − EK ) and so inward INa is absolutely larger than IK and
Inet would be inward. Thus, Inet is dependent on not only gK and gNa , but also on Vm , ENa and EK .
4. In conclusion Threshold is not a simple concept but rather is dependent on many factors.
5. When plotting out the INa it was noticed that the record of inward INa was not a smooth curve but
rather had a small dip in it when an A.P. was produced.
On the other hand the curve of g Na produced during an A.P. did not have such a dip. This suggests
that the other factor required to generate INa must be the source of the dip. That other factor is the
driving force term (Vm − ENa ). Indeed as the action potential nears its peak the Vm is approaching
ENa and hence INa becomes small. But as the A.P. begins to decay (Vm − ENa ) becomes larger again
and since gNa is still relatively large, INa becomes larger.
6. The Concept of Accommodation.
A rectangular pulse of current of amplitude 2.5 and lasting for 10ms was shown to be sufficient to elicit
an action potential.
However, a current ramp increasing over the course of 10ms from 0 to 3.4 did not elicit an A.P. Why
wasnt the larger stimulus more effective at eliciting A.P.s than the smaller stimulus?
7. Both stimuli produced depolarizations of roughly equal magnitude. These epolarizations activate gNa
by increasing the m parameter but they also decrease the h parameter. But the h parameter is relatively
slow to change, so the rapid onset of the rectangular pulse of current activates m rapidly but does not
cause much change in h.
On the other hand the current ramp depolarizes Vm relatively slowly so that m is slowly increased but
h is also slowly decreased. As a result of this decrease in h, gNa does not increase as rapidly as it does
when the rectangular pulse of current is applied and so INa does not increase sufficiently to make Inet
inward.
8. The concept of Refractoriness
A pulse of current of amplitude 100 and lasting 0.1ms elicited an A.P. but if this is followed 8ms later
by another current pulse of 100 and lasting 0.1ms no A.P. was produced only a small depolarization.
This is the phenomenon referred to as Refractoriness.
If the second current pulse is increased to 500 for 0.1ms, then a second A.P. is produced but the
amplitude of this second A.P. was about 20mV smaller than the amplitude of the first A.P.
Since an A.P. was produced by the stronger second stimulus 8ms after the first stimulus, 8ms would
be in the axon’s “relative refractory period.”
9. What factor(s) produce this refractoriness?
It was suggested that the IK was still elevated 8ms after the first stimulus and that this would make it
harder to depolarize the membrane above Threshold. This was suggested because at 8ms the Vm was

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 still slightly hyperpolarized as part of the afterhyperpolarization. Looking at the value of gK at 8ms
showed that it was increased over resting levels, so this increased IK is certainly a factor.
However, it was also observed that gNa did not increase after the second stimulus as much as it did
after the first stimulus which suggests that the second stimulus does not produce as large an inward
INa as the first stimulus. The reason for this decrease in gNa was investigated by plotting out the m,
n, and h factors after the first A.P.
It was found that not only was n higher at 8 ms than at rest but also h was smaller than at rest. Thus
there is a higher gK at the time of the second stimulus there is also a smaller gNa due to the smaller
value of the h factor. Thus both increased K channel activation and decreased Na channel opening
(also referred to as increased Na channel inactivation) decrease the ability of the membrane to produce
a second A.P.
10. The concept of Anodal Break
It was demonstrated that after the membrane is hyperpolarized for a while (i.e., by a stimulus of -10
last in for 5 ms) that the transmembrane gradually returns toward resting potential but then overshoots
resting potential and an A.P. is elicited. This phenomenon is called “Anodal Break.”
11. What factors produce Anodal Break?
By running the -10 stimulus for 5ms and eliciting an A.P. while also plotting gNa and gK during the
course of this stimulation and the elicited A.P., it was noted that during the A.P. elicited that gNa
rose to a value of about 50, whereas for the usual A.P. elicited by depolarization the gNa rose only to
a value of 40. Thus after the hyperpolarization gNa was larger than usual for a period of time.
12. What factor(s) might produce a larger gNa ? The factors controlling gNa are m and h so the program
which plots these variables was then used when the -10 stimulus lasting 5 ms was applied. It was
seen that during the period of hyperpolarization h gradually increased from a value near 0.65 to about
0.85. When the hyperpolarizing current was stopped h started to decrease slowly toward 0.65, but
remained considerably higher than 0.65 at the onset of the anodal break A.P. Thus, the increase in the
inactivation parameter h produced a larger gNa which in turn led to more inward INa resulting in an
inward I Net which was sufficient to elicit an A.P.

4 Is this a “good” model?

1. A “good” model is one which accurately describes the phenomena that the designers of the model set
out to describe.
2. The Hodgkin-Huxley model describes the properties of an A.P. very effectively.