Cell Physiology: IONIC EQUILIBRIUM

CTS/Physiology Lecture 16 Tuesday, October 8, 2002 9:10-10:00 Understanding what it means for an ion to be in equilibrium will help you to understand the basis for resting membrane potentials, action potentials, and synaptic transmission Reading: Berne & Levy, Physiology, ed. 4, pages 21-26 or Berne & Levy, Principles.., ed. 3, pages 19-24 Self-instructional Package: Course Website Questions: kutchai@virginia.edu

Ionic Equilibrium: Learning Objectives

Define the electrochemical potential () of an ion. Write the equation for the difference () between the electrochemical potential of an ion on one side of a membrane and that on the other side of the membrane. What does it mean? Use the Nernst Equation to determine whether an ion is in equilibrium. If the ion is not in equilibrium, determine in which direction the ion will tend to flow. Compute the electrical potential difference (voltage) across a membrane that is permeable to only one ionic species. Identify the Gibbs-Donnan Equilibrium and explain its implications for distribution of ions and water between cytosol and extracellular fluid

Which way will an ion flow spontaneously?

In situation #1 Na+ will tend to diffuse from A to B due to both the concentration difference and the electrical potential difference. In situation #2 Na+ will tend to diffuse from A to B due to the concentration difference and from B to A due to the electrical potential difference. In #2, which way will Na+ diffuse? We cannot tell!

How do we know which way an ion will diffuse?

Here Na+ tends to diffuse from A to B due to the concentration difference, but from B to A due to the electrical potential difference. Which of these two tendencies is larger? We need to be able to quantitatively compare the concentration force with the electrical force. The electrochemical potential allows us to do this.

What is the electrochemical potential?

The electrochemical potential () of an ion is defined as = o + RTlnC + zFE, where o is the electrochemical potential in some reference state R is the ideal gas constant T is the absolute temperature C is the concentration of the ion z is the valence of the ion F is Faradays Number (96,500 coulomb/mole) E is the electrical potential (e.g. in volts)

What is the meaning of electrochemical potential?

= o + RTlnC + zFE has units of energy/mole o is the electrochemical potential in some reference state, say at 1 M concentration at 20oC RTlnC is the energy 1 mole of ions possesses due to their concentration zFE is the energy 1 mole of ions possesses due to the electrical potential

What is the meaning of ?

What is the difference in the electrochemical potential of an ion across a membrane?
A = o + RTlnCA + zFEA B = o + RTlnCB + zFEB = Na(A) - Na(B), so that
= RT ln C( A ) + zF(E A E B ) C(B )

= RT ln

C( A ) + zF(E A E B ) C(B )
remember this equation

The first term: RTln(CA/CB) is the difference in energy between a mole of ions on side A and side B due to the concentration difference The second term: zF(EA - EB) is the difference in energy between a mole of ions on side A and side B due to the electrical potential difference A positive value of indicates greater electrochemical potential on side A than on side B. A negative value indicates greater energy on side B than on side A

C( A ) + zF(E A EB ) C(B ) An ion will tend to diffuse from where its is higher to where its is lower. = RT ln

What is the meaning of ?

and the energetics of transport

When an ion flows from higher to lower , it releases energy. This energy can be harnessed to do work. How much work? At most, for each mole of ions that flows. When an ion is transported from lower to higher , it requires energy. Work must be done. How much work? At least, for each mole of ions that flows.

RTln(CA/CB) is the tendency to diffuse from A to B because of the concentration difference: the concentration force. zF(EA - EB) is the tendency for the ion to diffuse from A to B due to the electrical potential difference: the electrical force A positive value of either force indicates a tendency for the ion to diffuse from A to B. A negative value indicates a tendency to diffuse from B to A.

What does it mean for an ion to be in equilibrium?

at equilibrium:
= RT ln C( A ) + zF(E A EB ) 0 C(B )

The Nernst Equation

Solving
= RT ln C( A ) + zF(E A E B ) = 0 C(B )

This can happen if there is no concentration difference and no electrical potential difference This happens more generally when the concentration force is equal and opposite to the electrical force When an ion is in equilibrium between side A and side B: Its electrochemical potential on side A is equal to that on side B There is no net force on the ion There is no net spontaneous flow of the ion

For EA-EB E E = RT ln C( A ) = RT ln C(B ) A B zF C(B ) zF C( A ) gives This is called the Nernst Equation It allows us to calculate the electrical potential difference, EA - EB, that just balances a particular concentration ratio The Nernst Equation holds ONLY for an ion that is in equilibrium Any ion that is in equilibrium will satisfy the Nernst Equation

Using the Nernst Equation Using the Nernst Equation

E A EB = RT C( A ) RT C(B ) ln ln = zF C(B ) zF C( A )

K+ is in equilibrium across the membrane What is EA EB?

EA EB = 60 mV 60 mV C(A ) 0 .1 log log = z C (B ) +1 0 . 01

ln(x) = 2.303 log(x) 2.303RT/F 60 mV

EA C(A ) 60 mV C (B ) 60 mV log log EB = = z C (B ) z C(A )

EA EB =

60 mV log( 10 ) = ( 60 mV )( 1 ) = 60 mV +1

60 mV will just balance a 10-fold concentration ratio!

Using the Nernst Equation Is Cl- in equilibrium across the membrane?

A membrane separates chambers A and B. Which of the following statements is correct? A. Cl- will flow spontaneously from B to A. B. K+ will flow spontaneously from A to B. C. The maximum amount of energy released by one mole of Clions flowing from A to B is equal to RTln(10) + (96,500)(0.06). D. The minimum amount of energy required to transport one mole of K+ from B to A is RTln(10) + (96,500)(0.06). E. Both K+ and Cl- are in equilibrium.
EA EB = 60 mV 60 mV 1 C(A ) = log log 1 0 .1 C (B ) z

E A EB =

60 mV log( 10 ) = ( 60 mV )( 1 ) = + 60 mV 1

If EA - EB were +60 mV, the electrical force would just balance the concentration force. So, EA-EB is larger than it needs to be, so that the electrical force is greater than the concentration force. Thus Cl- will flow from B to A

Using the Nernst Equation

The Nernst Equation tells us what the membrane potential would have to be in order for an ion to be in equilibrium The Nernst Equation gives us a theoretical number Frequently we can measure the membrane potential The measurement gives us a real number Comparing the Nernst potential with the actual membrane potential tells us important things

Using the Nernst Equation

If E measured = E calculated (from the Nernst Eqn, that is the equilibrium potential for the ion), the ion is in equilibrium If E measured is same sign as E calculated, but smaller, then concentration gradient determines direction of flow If E measured is same sign as E calculated, but larger, then electrical potential difference determines direction of ion flow If E measured is opposite in sign to E calculated, then both electrical force and concentration force are in same direction. The ion cannot be in equilibrium.

Gibbs-Donnan Equilibria
Cytosol and the extracellular fluid contain some ionic species to which the plasma membrane is permeable and others to which the plasma membrane is impermeable Cytosol has a significant concentration of charged, impermeant macromolecules, such as proteins and nucleic acids. These are predominantly anionic (negatively charged) Many intracellular metabolites are also impermeant anions The presence of impermeant species influences the equilibrium distribution of permeant species

a Gibbs-Donnan model situation

The membrane is permeable to K+ and Cl-, but impermeable to X-. Initially K+ is in equilibrium, but Cl- has a tendency to flow from B to A As Cl- flows from B to A, a membrane potential (side A negative) builds up. This provides a driving force for K+ to flow as well from B to A

When will Cl- and K+ stop flowing?

When they reach equilibrium! What will characterize equilibrium for both K+ and Cl-? Both will satisfy the Nernst Equation.
K = RT ln Cl = RT ln [K ] A + F( E A E B ) = 0 [K ]B [Cl ]A F( E A E B ) = 0 [Cl ]B

What will characterize this equilibrium?

K = RT ln Cl = RT ln [K ] A + F(E A E B ) = 0 [K ]B [Cl ]A F(E A E B ) = 0 [Cl ]B

Add the two equations, divide by RT, take antilogs of both sides, cross-multiply, and we obtain

[K]A[Cl]A = [K]B[Cl]B
This is known as the Donnan Relation or the Gibbs-Donnan Relation Note that satisfying the Donnan Relation simply means that both ions are in equilibrium

How much Cl- and K+ will flow?

A slightly larger number of Cl- flows from B to A than K+ However, in chemical terms the difference between the number of Cl- and K+ ions that flow is insignificant The Law of Electroneutrality: In any macroscopic volume of solution the number of + and charges must be equal

What will characterize this equilibrium?

Assume A and B have equal volumes If the decrease in [Cl-] and [K+] in B are equal to Z, Then [K+] and [Cl-] increase by Z in A [X-] cannot change [K]A[Cl]A = [K]B[Cl]B (0.1 + Z)(Z) = (0.1 Z)2 So Z = 0.0333

And the equilibrium concentrations are

Note that there is higher [K+] in A than in B There is higher [Cl-] in B than in A If K+ and Cl- are in equilibrium, there must be a membrane potential KA/KB = ClB/ClA = 2 Both predict EA - EB = -18 mV Note that electroneutrality prevails in both A and B

What about osmotic balance?

Note that the sum of [K+] and [Cl-] in A is greater than their sum in B There is [X-] present in A, but none in B Consequently, there is a higher concentration of osmotically active solute in A than in B Unless it is restrained in some way, water will flow from B to A

How do cells cope with the osmotic consequences of Gibbs-Donnan equilibria

The membrane is permeable to all ionic species except Y-. We do not know the value of the membrane potential. Which of the following statements is correct? A. K+ and Cl are definitely in Gibbs-Donnan Equilibrium. B. It is not possible for K+ and Cl to be in Gibbs-Donnan Equilibrium. C. It is not possible for Na+ and Cl to be in Gibbs-Donnan equilibrium. D. There is no value of membrane potential that would allow both K+ and Cl to be in equilibrium. + and Cl may be in Gibbs-Donnan equilibrium. E. Na

Plant cells have a rigid cell wall and develop turgor pressure Animal cells pump certain other ions out of the cytosol to compensate The principal ion that is pumped is Na+ Poisoning the Na,K-ATPase causes cells to swell The Na,K-ATPase is important in regulation of cell volume

Features of Gibbs-Donnan equilibria

The side with the fixed anions (corresponds to the cytosol) has: Greater concentration of mobile cations Smaller concentration of mobile anions Negative membrane potential Greater osmotic pressure