Neurophysiology – lecture 16

March 3, 2011

1 Review:
1. Last Tuesday we took a complex pattern of currents produced by a squid axon being recorded in a
voltage clamp when the transmembrane potential was stepped to a voltage between -40 and = +40mV
and broke this pattern into 4 separate components.
2. It should be pointed out that one can only measure currents if a voltage clamp is being used.
3. Using the current record shown in Figure 10A as an example. We adopted the general model for 4
independent currents: IT = Ic + IL + INa + IK
4. A small voltage step of +10mV produced only an initial outward spike of current followed by a small
and prolonged outward current. We could identify these as currents generated initially across the
membrane capacitor, Ic , and later as current across the membrane resistance, IR . Both this capacitance
and resistance are assumed to be unmodified by changes in the transmembrane potential, Vm , so a
voltage step in the opposite direction equal to -10mV amplitude produces an Ic and IL of the same
magnitude but opposite polarity as the voltage step to +10mv. Hence these 2 current records when
added together produce a current record in which no current is visible.
5. Using this observation as a basis, every depolarizing voltage step used is followed by a hyperpolarizing
voltage step of the same magnitude. When the current records of these 2 voltage steps are added
together, the Ic and IL components of the total I are removed leaving only INa + IK . This procedure
simplifies the current record considerably to equal just INa + IK .
6. We then noted that a voltage step to ENa would produce a current record which only contained IK
since INa = 0 at ENa . In the case of the current records shown in Figure 10A, the voltage step to
+52mV (ENa ) must therefore contain no INa and therefore must be only IK .
7. To obtain current records of IK at other transmembrane potentials we can change [Na]o and thereby
change ENa . A voltage step to this new ENa will then produce a current trace at the transmembrane
potential which corresponds to ENa . By changing [Na]o over a range of concentrations an entire family
of IK curves can be generated representing the IK produced by voltage steps to different transmembrane
potentials.
8. If a current record is obtained at these same transmembrane potentials when the squid axon is bathed
in seawater this current record will contain both INa and IK . Consequently, if the current record
obtained in seawater by a voltage step to voltage X (therefore a current record containing both INa
and IK ) is subtracted from a current record produced by the same voltage step but obtained from
the axon while it is bathed in a “seawater” medium whose [Na]o has been reduced to produce an ENa
equal to voltage X (therefore a current record containing only IK ), the resultant current record will
contain only INa .

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 9. Following this strategy and using a variety of different [Na]o concentrations it is possible to generate a
family of INa records each produced at a different transmembrane potential.
10. This family of INa records can be converted to conductance curves by use of the equation
INa
gNa =
Vm − ENa
to generate a family of gNa curves as portrayed in Figure 11A.
11. Likewise, the family of IK records can be converted to conductance curves by use of the equation
IK
gK =
Vm − EK
to generate a family of gNa curves as also portrayed in Figure 11A.
12. Note that in Figure 11A the open circles represent data points obtained by the procedure outlined
above, while the dark line was generated by the equations that Hodgkin and Huxley developed to
describe these data points.

2 Properties of gNa and gK

1. If the maximum values obtained in the gK records at each voltage studied are plotted against the Vm at
which the current record was obtained, then a graph like that of Figure 11B is produced. Note that this
record has a very steep rising phase beginning at about -30mV and then plateaus at transmembrane
potentials above +30mV.
2. Similarly, if the peak values of the gNa curves are plotted against the Vm at which the current record
was obtained, then we see a very similar steep rising phase beginning near -30mV and a plateau at
transmembrane potentials above +30mV.
3. These graphs document the voltage dependent character of the Na+ and K+ ion channels.

3 Resynthesizing A.P.s
1. Knowing the voltage dependence and time course of INa and IK under voltage clamp conditions,
Hodgkin and Huxley then attempted to synthesize this information into a mathematical model of how
action potentials are produced when the transmembrane potential is not clamped but rather left free
to change.
2. They began by assuming an axon at its resting potential, Vrest , is subjected to a slight depolarization,
Vm1 resulting in a transmembrane potential of Vrest + V + m1 .
3. To make the conductance curves of Figure 11A applicable, we will assume that this depolarization
raises the transmembrane potential to -39mV. This change in transmembrane potential after 0.2ms
will increase gNa slightly but have no apparent effect on gK . 4) The slight increase in gNa will increase
INa since
gNa
INa =
Vrest + Vm1 − ENa
This INa will be inward since the driving force term in this equation is negative.
At the same time (0.2ms) IK will not increase much if at all because gK does not appear to have
increased.

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 4. Therefore, the Inet = INa + IK will be small but inward.
5. This inward Inet · 0.2 ms will produce an increase in intracellular +dq (Inet · dt = dq).
6. This dq will be distributed uniformly around the inside of the surface membrane because of the repulsive
forces present between like charged ions.
7. In this position these surface charges will influence charges on the other side of the membrane and this
will create a small outward capacitative current.
dq
8. This increase in + surface charge will also change the transmembrane potential, since = Cin and
dVm
dq
therefore dVm = . Call this small change in transmembrane potential will equal Vm2 . So now the
Cin
transmembrane potential equals Vrest + Vm1 + Vm2 .
9. This last step brings us back to step 2 above will a slightly higher transmembrane potential which will
produce a slightly larger gK at 0.4ms with still almost no change in gK , etc.
10. Thus, one can progress through steps 2 to 10 again, and again, and again. As one progresses through
successive loops gK begins to rise and then gNa begins to fall due to inactivation, etc. But eventually,
after progressing through about 500 complete cycles of calculations the entire waveform of a squid axon
action potential is generated by the transmembrane potentials values that are calculated.

4 Properties of this cycle of calculations

1. First note that this is a cycle. The initial input to the cycle, a slight depolarization, results at the end
of the cycle in an additional depolarization. Thus what initiates the cycle winds up producing more of
what initiated the cycle. A cycle of this type is known as a “positive feedback loop.”
2. It is this positive feedback aspect of the cycle that results in the A.P. being “All-or-None.” The initial
depolarization will result in an output that goes to completion or saturates unless some other process
holds it in check.
3. There are very few positive feedback cycles in biology under normal physiological conditions. If we
think of other positive feedback devices – like the atomic bomb – I think we can see why.
4. But of course the amplitude of the action potential is regulated so there must be some controls present.
Any process that is regulated achieves that status by virtue of some negative feedback processes.
Negative feedback occurs when the output of a process – like the transmembrane potential in this case
- is feedback to the input to reduce subsequent output.
5. In the case of A.P.s there are 4 sources of negative feedback:
(a) As Vm becomes more depolarized, (Vm − ENa ) becomes smaller and so INa becomes smaller.
(b) As Vm becomes more depolarized, (Vm − EK ) becomes larger and so IK becomes larger and more
effectively reduces the magnitude of the inward Inet .
(c) With the depolarization maintained for a longer period of time (1 ms), gNa begins to inactivate
which of course reduces gNa and hence INa .
(d) With the depolarization maintained for somewhat longer (0.8ms), gK begins to increase and hence
IK increases until later a net outward current is developed.
6. These negative feedback effects insure that the peak of the A.P. will never quite reach ENa and that
the Vm will repolarize after the A.P. peak.

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5 What determines “threshold?”
1. In the computer simulation A.P. we showed in the previous class we showed that for an axon at rest,
stimulus of 6.40 did not produce an A.P., but one of 6.41 did elicit an A.P. This type of observation
suggests that there is a strict threshold voltage above which A.P.s are generated and below which they
are not generated.
2. However, we also showed that a 5ms long hyperpolarization resulted in an A.P. when the stimulus was
removed. So the “threshold” cannot be strict.
3. So what determines the “threshold?”
4. The answer is that when the net inward ionic current, Inet, just becomes greater than 0, then the
membrane is at threshold for producing an A.P.
5. However, it should be noted that current only flows in closed circuits which means in this case that
the net inward ionic current is immediately returned to outside the cell as capacitative current. Hence,
when the outward capacitative current, Ic , becomes greater than 0 the cell becomes depolarized and
the sequence producing A.P.s begins.
6. The “catch phrase” to remember is: “Outward capacitative current depolarizes.”

6 Generation of the Hodgkin Huxley Mathematical Model

1. To facilitate the mathematical formulation of the processes underlying the A.P. Hodgkin and Huxley
assumed that gNa and gK had some maximum value. We now know that this maximum value would
be reached when all the Na+ or K+ channels in the axon membrane were opened. These values of the
conductances were designated gNamax and gKmax . To deal with the fact that these conductances are not
always at their maximal value they formulated:

gNa = k gNamax and gK = k gKmax where k ≤ 1

2. They fractions they chose were designed to fit the conductance data seen in Figure 11A.
These fractions are:
gNa = m3 h gNamax and gK = n4 gKmax
where m is the parameter controlling Na+ channel activation, h is the parameter controlling Na+
channel inactivation and n is the parameter controlling K+ channel activation.
3. In explaining the basis of certain A.P. properties we use these terms activation and inactivation in
referring to these parameters.