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Neurobiology 15-BIOL-540 Winter, 2012

The document discusses ion channels and their role in generating membrane potential. It notes that small changes in the number of ions crossing the membrane can significantly impact membrane potential. Voltage clamp experiments allow researchers to measure currents required to hold voltage constant and determine the properties of sodium and potassium channels. The patch clamp technique enables studying individual ion channels. There are many variants of potassium channels that differ in their behavior and cell physiology experiments involve manipulating ion concentrations and permeabilities to study their effects on membrane potential.

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60 views63 pages

Neurobiology 15-BIOL-540 Winter, 2012

The document discusses ion channels and their role in generating membrane potential. It notes that small changes in the number of ions crossing the membrane can significantly impact membrane potential. Voltage clamp experiments allow researchers to measure currents required to hold voltage constant and determine the properties of sodium and potassium channels. The patch clamp technique enables studying individual ion channels. There are many variants of potassium channels that differ in their behavior and cell physiology experiments involve manipulating ion concentrations and permeabilities to study their effects on membrane potential.

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bcastle1
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
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Neurobiology 15-BIOL-540 Winter, 2012 Lecture 5

It does not take many ions moving across the membrane to change Vm
100m
[K+]= 400mmol
Volume: 5.24x10-10L # K+ ions = 5.24x10-10L x 400 mmol/L x 6.02 x 1023 ions/mol = 1.26X1014 K+

1.47x108 K+ leak out


126,000,000,000,000 -147,000,000 125,999,853,000,000 Or 1/860,000 K+ ions

1.26x1014 K+

1.26x1014 K+

0mV

EK = -75mV

How do we know what happens during the six phases of action potentials?

Voltage clamp electrophysiology: holds voltage constant, measures current required to do this.

Command

Current (not voltage!)

Current sign indicates which way positive charges are moving (net).

This is the time-dependent pattern of current that occurs upon depolarization.

This is actually a composite of two separate currents: Na+ and K+.

We can tell this by blocking one channel, then the other.

We can tell this by blocking one channel, then the other.

Or by removing ions in the extracellular space.

Hodgkin & Huxley could measure macro-currents.

But they only guessed at the properties of channels.

TWO BROAD CATEGORIES OF CHANNELS

The invention in the 1970s of patch clamp electrophysiology.

The patch clamp technique

1. Current is inward. 2. Current reverses at ENa 3. Current decreases as function of [Na+] or TTX in bath. 4. Summed microcurrents match measured macrocurrents for Na+.

4. Summed microcurrents match measured macrocurrents (timing, dynamics).

Channel opening is probabilistic.

Similar lines of evidence connect microcurrents to K+ macrocurrents.

How do you study the electrical behavior of such diverse channels, and know which channel you are recording?

LOTS of different versions of each of these. These versions may differ in their behavior.

For instance, K+ channels:

For instance, K+ channels:

For instance, K+ channels (~100 K+ channel genes!!!).

Cell interior [A+]=10mM. [A+]=100mM. [B+]=100mM. [B+]=10mM. [C--]=100mM. [C--]=10mM. [D--]=10mM. [D--]=100mM. (Other ions, too)

The cell is at a resting potential of -58mV. The membrane potential Vm will ______ after the following changes in membrane permeability to ions A, B, C, D:

PA is increased. PB is decreased.

Cell interior [A+]=10mM. [A+]=100mM. [B+]=100mM. [B+]=10mM. [C--]=100mM. [C--]=10mM. [D--]=10mM. [D--]=100mM. (Other ions, too)

The cell is at a resting potential of -58mV. The membrane potential Vm will ______ after the following changes in membrane permeability to ions A, B, C, D:

pA is increased. pB is decreased.

Well discuss these in the context of cell-cell communication next week.

Channel structure: how can they be gated? Selective?

Bacterial K+ channel.

A look at the proteins responsible for generating and maintaining membrane potential.

Sodium potassium pump

Na+-K+ ATPase pump

Ca2+ pump

10

16

~100

several

Study dynamics of channels by expressing them ectopically in frog eggs.

Synaptic transmission

Criteria that define neurotransmitters.

Two families of neurotransmitters.

Neurotransmitters are released in discrete quanta.

Two families of postsynaptic receptors

Ligand-gated ion channels aka Ionotropic receptors

G-protein coupled receptors aka Metabotropic receptors

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