Membrane Potential 1

Cell Membrane as a capacitor

C = Q/V

1 Farad = 1 coulomb/1 volt

C = Q/V = koA/d

ko = dielectric constant, o = 8.854 x 10-12 F/m, k > 1

A = area of plates
d = distance plates are apart

Dielective constant is basically how effective of an insulator is the material between the
plates. Cell membrane is the insulator and the salt solutions inside and outside of the cell
are the plates.

Take home point: greater the distance (d), lower the capacitance.

Since the cell membrane is very thin, it can separate a significant charge per volt potential
across the membrane.
Membrane Potential 2

Nernst Equation

Two forces acting on ions (or any charged molecule) in solution crossing a permeable
membrane:

[ X ]A
1) Concentration Gradient: Force = RT ln
[ X ]B
R=gas constant, T=absolute temperature

x
x
x x x
x
x x
x

A B

2) Electrical Potential across the membrane: Force = zF(EA - EB)

F=Faraday’s number, z=charge of particle (e.g. +1, -1), E=electrical
potential

+ -
+ -
+ -
+ -

A B
Membrane Potential 3

Combining these two forces we arrive at the concept of:

Electrochemical potential (energy/mole) = RTln[XA/XB] + zF(EA - EB)

When RTln[XA/XB] + zF(EA - EB) = 0, electrochemical equilibrium has been achieved.

Electrochemical equilibrium occurs when electrochemical potential is zero, in other

words when concentration gradient = electrical gradient, with opposing directions.

RT [ X ] A
EA - EB = ln
zF [ X ]B

EA - EB = E (we will just call it E)

Reworking the equation converting it from natural log to ten base log and including
Faraday's constant, molecular gas constant and assuming physiological temperature of
310o K (37o C), the equation can be simplified to the following:

− 61mV [X]inside
E= log
z [X]outside
z=charge of ion

This is the Nernst Equation aka Reversal Potential Equation.

Note that this will give the electrical potential of the inside of the cell relative to outside
of the cell.

Your text has the equation expressed in the following form.

61mV [X]outside
E= log
z [X]inside

Please note that the 61 is positive because the "out vs in" concentrations ratio has be
reversed. Different texts have it either way. It doesn’t matter which formula you use as
long as you don't confuse the two forms.

The Nernst equation is the Equilibrium Equation for a charged solute.

Membrane Potential 4

We can plot the Nernst Equation for potassium. Note that the slope of the line is about 61
mV. So, for every 10-fold change in potassium concentration, there is about a 61 mV
change in potential. This would also be true for the other electrolytes in biological
systems, e.g., sodium etc. The “61” is a function of RT, z, F and conversion from ln to
log.

You will also notice that there is no factor in the equation for permeability of charged
particles relative to the membrane. Permeability is irrelevant because we are talking
about an equilibrium state. Permeability will only affect the time it takes to reach
equilibrium.
Membrane Potential 5

Example 1. What electrical potential would be required to maintain this concentration

gradient? (assumption: membrane is only permeable to K+, i.e., not water.)

0.1M 0.01M

K+ K+

inside outside

Example 2. Suppose that the electrical potential was +100 mV in the inside. Would
the ions flow from one side to another? Which side?

HCO3- HCO3-

1M 0.1M

inside outside
Membrane Potential 6

Nernst Equation can be used to predict the direction that ions will flow.

As stated earlier, the Nernst value is the theoretical potential at which the electrical and
concentration forces acting on the ion are equal and opposite, which means no net
movement for that ion across the membrane.

If the potential is above the Nernst value, the ion will flow in one direction. If the
potential is below the Nernst value, the ion will flow in the opposite direction.

Since the Nernst value is the “tipping point” shall we say, as to which direction the ions
will flow across the membrane, we use often use the term “reversal potential” as another
term for the Nernst value.

The last few examples we have been “putting” a potential across the membrane with a
battery when illustrating the Nernst value effect on flow or charge.

In real biological systems, why would there be an uneven distribution of charge across a
cell membrane creating potential across cell membranes?
Membrane Potential 7

In biological systems there are multiple factors that creates a potential across a
membrane. Two major factors are:

1) Higher concentration of negatively charged proteins inside the cell, which the cell
membrane is impermeable to, compared to outside the cell.

2) Ion pumps that cause a net change in potential across the membrane. (more on this
later)

Donnan effect: The presence of charged particles that cannot cross a semipermeable
membrane in a system where there are other charge particles that can cross the
membrane will reach an equilibrium state such that there is an unequal distribution of
charge resulting in an electrical potential across that membrane. The equilibrium is
called the Donnan Equilibrium, also known as the Gibbs-Donnan Equilibrium.

[K+]=0.1M [K+]=0.1M

[Proteins-]=0.1M [Cl-]=0.1M

A [inside cell] B [outside cell]

The above are initial conditions. Suppose the membrane is not permeable to proteins- but
permeable to K+ and Cl-, what will happen to the concentrations?

Two forces acting on charge particles:

1) Concentration gradient force

2) Electrical force

[K+]=0.1333 M [K+]=0.0666 M

[Proteins-]=0.1M [Cl-]=0.0666 M

[Cl-]=0.033 M

A [inside cell] B [outside cell]

Double Donnan Effect – impermeable charged particles on both sides of the membrane.
Membrane Potential 8

If Donnan equilibrium is achieved, then the Nernst values for each ion should be equal.

For K+ [K+]=0.1333 M [K+]=0.0666 M

− 61mV [KA] [Proteins-]=0.1M [Cl-]=0.0666 M

E= log
+1 [KB]
[Cl-]=0.033 M
0.1333
E = −61mV log
0.0666 A [inside cell] B [outside cell]

E = −61mV log 2
E = −61(0.3)
E = −18mV

For Cl-

− 61mV [ClA ]
E= log
−1 [ClB]

0.0333
E = 61mV log
0.0666

E = 61mV log 0.5

E = 61( −0.3)
E = −18mV

The Nernst equation only applies to systems where there are some charged particles that
cannot cross the membrane.

Add up the charge in chamber A and then chamber B. What do you get?

What about osmolarity?

Membrane Potential 9

Electrogenic pumps

Both the Donnan effect and electrogenic ion pumps play roles in maintaining a potential
across a cell membrane.

Resting Membrane Potential (RMP)

Assume we have a cell with a resting membrane potential and has the following ion
distributions at that resting membrane potential.

145 mM Na+
15 mM Na+
140 mM K+
5 mM K+
10 mM Cl-
110 mM Cl-
Proteins (-)
infinite volume

Do we have a Gibbs-Donnan equilibrium? Why?

140
K+ : E = −61log = −61log 28 = −61(1.45) = −88.3mV
5

10
Cl- : E = +61log = 61log 0.09 = 61(−1.04) = −63mV
110

15
Na+ : E = −61log = −61log 0.103 = −61(−0.98) = +60mV
145

Why do we have so much sodium in ECF and so much potassium in the ICF?
Membrane Potential 10

145mM 5mM 110mM

15mM 140mM 10mM

Na+ K+ Cl-
measured E = -72 mV
Proteins-

The measured resting membrane potential is -72 mV

If the cell were in Donnan equilibrium, the Nernst equation for each ion should yield the
measured membrane potential value of -72mV.

140
K+ : E = −61log = −61log 28 = −61(1.45) = −88.3mV
5

10
Cl- : E = +61log = 61log 0.09 = 61(−1.04) = −63mV
110

15
Na+ : E = −61log = −61log 0.103 = −61(−0.98) = +60mV
145

Knowing the ion concentrations in and out of the cell, can you predict the resting
membrane potential?

Answer is yes, but there is another variable we must consider.

Conductance
Membrane Potential 11

Since the membrane is not equally permeable to these 3 ions we can give a "weighted"
figure for each ion conductance. In other words, we can factor in a "g" value for each ion.

Chord Conductance Equation:

gKEK gN aEN a gClECl

Em = + +
g g g

g = 1/R

R=V/I
Amps
g = mho =
Volts
1 mho = 1 siemen; some texts use siemens instead of mhos.

conductance (g) in expressed in mho/cm2

g = (gK +gNa + gCl)

Cell membranes are not equally permeable to all ions. Na+ and K+ cross the cell membrane
via Na+/K+ "leak channels". These leaky channels are far more permeable to K+ than Na+.

When ions are distributed across a membrane, and are removed from electrochemical
equilibrium, each ion will tend to force the transmembrane potential towards its own
equilibrium potential, as calculated from the Nernst equation. The more permeable the
membrane to a particular ion, the greater strength that ion will have in forcing the membrane
potential toward its equilibrium potential. Since normal resting cell membranes are 10-100x
more permeable (which usually means conductance is greater) to K+ than Na+, K+
concentration gradient is the most influential factor in the resting membrane potential.

There is another common equation used to calculate resting membrane potentials with given
ion concentrations. It is expressed in terms of "permeability". Permeability is dependent on
the nature of both the ion and the membrane. Permeability in this equation is expressed in
terms of each ion’s ability to move across a membrane relative to the other ions. It's called
the Goldman Equation. It is also known as the Goldman-Hodgkins-Katz Equation. It's
expressed at physiological temperature:

Pk[K +]o + PNa[ Na+]o + PCl[Cl−]i

Em = 61log
Pk[K +]i + PNa[ Na+]i + PCl[Cl−]o

Typical permeability ratios for a mammalian neuron is K:1.0, Na:0.04, Cl:0.45 (FYI)

Note that if the permeability of Na and Cl were zero, then this equation becomes the
Nernst equation for K.
Membrane Potential 12

Conductance vs Permeability.
These two concepts are often confused. While they can be related to each other they are
not the same. Conductance is an electrical concept. It must involve the movement of
charge and is a function of Ohm’s law. Permeability is a chemical concept which
involves the movement of any molecule, water, gas etc. regardless of charge, across a
membrane.

Let’s talk more about conductance and Ohm’s law. g = I/V

We can plot this on a graph.

Conductance is the slope of the line. For a given cell membrane with a fixed permeability,
g is a constant. As the function suggests, for a given cell membrane, if you know either
the I or the V, then you can predict the other if you know the g (slope). You can
empirically figure out the g by collecting data: put a potential across a membrane and
then measure the amperage. Do this for multiple voltages to get multiple amperage
readings and then the line appears, which give you the slope, i.e., conductance (g). Cell
membrane in real biological systems do not have a fixed permeability. If you alter the
permeability then you alter the g, which means you alter the slop of the line.

Looking at the equation g = I/V, what if there is no current and voltage, i.e., I = 0, V=0, is
g then equal to 0? Answer: no. The concept of conductance or resistance only makes
sense in the context of a hypothetical non-zero value of amperage and voltage. If
conductance is 0 or resistance is infinite, the there is no mathematical relation between I
and V. Ohm’s law does not apply. Also, if I is 0 then V must be 0 and 0/0 is
“indeterminate”.

If I were to hold a wire up in my hand and the wire is not attached to anything, and say
the resistance of this wire is x ohms, what I’m really saying is, if this wire’s resistance is
x ohms, then if you hook the wire up to a battery and I know the voltage of the battery, I
can now predict the amperage that will flow through the wire.
Membrane Potential 13

Real life Cell Membrane Potentials

In the real biological systems, cell membranes are permeable to various ions to various
degrees and this is dependent on which type of cell we’re taking about. Resting
membrane potential varies greatly, dependent cell type. Typical resting membrane
potentials for skeletal muscle is -95 mV, smooth muscle is -60 mV, neurons, -60 to -70
mV, red blood cells, -8 mV.

Ion Pumps

In real situations cell membranes have ion pumps moving molecules across the
membrane, Na+, K+, Cl-, Bicarbonate, Ca+2 pumps etc. The presence of ion pumps is a
major factor that determines the distribution of ions and resting membrane potential.
Pumps are continuously moving ions and at the same time ions are continuous leaking
in/out of the cell. Movement of charge means you have a current (amperage) therefore
Ohm’s law applies.

Because of the pumps and leaking ions in/out of cells, a resting membrane potential is not
in static chemical equilibrium, but we would say the membrane is in dynamic chemical
equilibrium or “Steady State” relative to movement of charge.

The Na+ / K+ATPase (electrogenic) pump plays a critical role in the resting membrane
potential. It pumps out 3 Na+ for every 2 K+ in, with a net gain of 1 ion pumped out of the
cell for each ATP hydrolyzed.
Membrane Potential 14

What would happen if you poisoned these pumps?

Sodium and Potassium gradients would become less because there is nothing to counter
the continual leaking of potassium out and sodium in. Other pumps are dependent on
these gradients (potassium and sodium) to move other ions and non-ions in or out of the
cell. This would also cause issues with osmotic pressure.

Ouabain is a compound that selectively inhibits some isoforms of the Na+ / K+ATPase
pump. It's used in experimentation to investigate the function of that pump.

The 3/2 pump ratio of sodium out and potassium in simply maintains normal ion
concentration gradients. The activity of this Na/K ATPase pump, because of its net
movement of +1 charge out of the cell, typically adds an additional degree of negativity
to the resting membrane potential, i.e., makes RMP to be more negative.

Typical ion concentrations in mammalian neurons (note that this can vary greatly,
depending upon the cell type:

Ion Inside Outside

Sodium 5-15 mM 140 mM
Potassium 140 mM 5 mM
Calcium 1 x 10 -4 mM 1-2 mM
Chloride 4-30 mM 110 mM

Palytoxin: produced by some types of coral and other marine planktons.

 Membrane Potential 15

Problems

At equilibrium

Infinite vol

____ Em

100mM K+
2mM K+

At equilibrium
Infinite vol

____ Em

2 M K+
40mM K+
 Membrane Potential 16

At equilibrium

Infinite vol

____ Em

8 mM Ca+2 4 mM Ca+2

Infinite vol

____ Em
g=20 mmho 100 mM
10 mM K+ K+
150 mM Na+ g=2 mmho 5 mM Na+
40 mM Cl- g=1 mmho 2 mM Cl-
 Membrane Potential 17

Infinite vol

____ Em
g=20 mmho 10 mM K+
10 mM K+ g=2 mmho 5 mM Na+
150 mM Na+ g=10 mmho 2 mM Ca+2
40 mM Ca+2

What potential would have to be in A relative to B for this system to be in equilibrium?

Assume A is inside the cell and B is outside the cell for the problems below.

3 mM 10 mM
K+ K+

A B
Membrane Potential 18

Which direction will sodium flow?

5 mM 100 mM
Na+ Na+

+50 mV

A B

Which direction will chloride flow?

120 mM 15 mM
Cl- Cl-

-30 mV

A B