Neuroscience Winter 2016

Lab # 2: Action potential and transmission of nerve

impulses
Objectives of the lab:
- Understand the differences between resting potential and action potential at the
molecular level.
- Understand how impulses are transmitted.
Introduction:
In lab # 1, you learned that the nervous system was made of the central nervous system and the
peripheral nervous system. You also learned about the types of cells found in those systems.
In lab # 2, you will learn how the two systems are connected and how impulses are generated, traveled
along the axons and are transferred from one neuron cell to the next.
Neurons are the cells responsible for generating and conducting electrical impulses from one part of
body to another as outlined and shown in the pictures below:
1. Sensory unipolar neurons in the PNS
are specialized in responding to
stimuli. They transmit info about
stimulus to the CNS in form of
electrical impulses.
2. Multipolar interneurons within CNS
transmit impulses between
components of CNS. Interneurons
receive input from sensory neurons,
integrate information, and influence
the functioning of other neurons.

3. Motor multipolar neurons of the

PNS transmit impulses away from CNS.
Carry the nervous system's output,
still in the form of electrical impulses,
to all other tissues and organs of the
body.

Neurons are therefore the functional unit of nerves. They are made of three sections:
1. Dendrites receive information from another cell and transmit it to the cell body
2. Cell body contain nucleus, mitochondria and other organelles
3. Axon conducts messages away from the cell body

Understanding the Transmission of Nerve Impulses

Neurons send messages electrochemically, meaning that chemicals (ions) cause an electrical signal that
is then transmitted from neuron to neuron as an electric current.
A few Principles of Electricity:
Opposite charges attract and energy must be used to separate them
Conversely when opposite charges come together, energy is liberated
Therefore separated electrical charges have potential energy
Measure of potential energy = voltage
Voltage is always measured between two points and is called the potential difference
Flow of electrical charge from one point to another is called current
The amount of charge moving between two points depends on voltage and resistance
(hindering of flow of charge)
Ohm's law states that "current = voltage/resistance"
Current is proportional to voltage (the greater the voltage, the greater the current) and is
inversely proportional to resistance (the greater the resistance, the smaller the current)

In the body, electrical currents are generated due to the flow of ions across cellular membranes and
resistance is provided by the membranes themselves.
The important ions in the nervous system are sodium and potassium (both have 1 positive charge, +),
calcium (has 2 positive charges, ++) and chloride (has 1 negative charge, -). There are also some
negatively charged protein molecules that play a role in the electrical balance between the inside and
outside of the cell. Ions move along chemical gradients (due to diffusion) and along electrical gradients
(move toward an opposite charge); therefore ions flow along electrochemical gradients.
It is also important to remember that nerve cells are surrounded by a membrane that is semipermeable and allows some ions to pass through while blocking the passage of other ions. Nerve cells
are unique in their capacity to manipulate the flow of charged ions across the cell membrane. They will
open or close channels for specific ions in response to a stimulus. The combined action of all these
channels generates a wave of voltage change that can travel throughout the cell and be transmitted to
neighboring cells.

A variety of ion channels occur in the plasma membranes and are classified as being either
passive or active
Passive (leakage) channels are always open
Active (gated) channels are made up of one or more proteins capable of undergoing changes to
open or close; some examples:
Chemically gated channels - respond to neurotransmitters
Voltage-gated channels - respond to membrane potential changes
Other gated channels - mechanical, pressure, light

Resting Membrane Potential Diagram

Plasma membrane in a neuron has an unequal distribution of ions and electrical charges between the
two sides of the membrane:
1. Outside is positively charged
2. Inside is negatively charged
This difference, known as resting potential (axon is not conducting an impulse), has a voltage of ~ -6070 mV. That is, 70 millivolts more negative inside the cells compared to the extracellular fluid
surrounding it. Note that different neurons have slightly different resting potentials. We say that the
cells are polarized. The resting potential is due to the fact that the inside of the cell contains negatively
charged proteins that cannot cross the membrane and also depends on the different equilibrium
potential of each of the main ion players (Na+, K+ and Cl-). An equilibrium potential is the potential at
which the two competing forces on the ions (a diffusional force, carrying the ion down its concentration
gradient, and an electrical force, moving the ion toward areas of opposite charge) are equal and
opposite. This electrochemical gradient across the cellular plasma membrane is a means of storing
energy that the cell can use in exergonic reactions.
Lets now consider the distribution of the following ions at equilibrium:

1- Na+ ions: At rest, sodium

ions are more concentrated
outside the neuron than inside.
This goes against both
diffusional and electrical
forces. So sodium ions will leak
back into the cell via non-gated
Na+ channels. However, the
unequal concentration of
sodium between the inside and
outside is maintained by
actively transporting the ions
against their concentration
gradient via the ATPdependent sodium/potassium
pump, which carries three
sodium ions out while bringing
two potassium ions in.
2- K+ ions: At rest, potassium
ions are more concentrated
inside the neuron than outside.
This is in accordance with the
electrical force as the inside of
the cell is negatively charged
but is against the diffusional
force. Non gated potassium
channels will therefore allow
movement of potassium ions
outside the cells but the
build-up of potassium inside
the cells against its
concentration gradient is
restored by the ATP-dependent
sodium/potassium pump
mentioned above. The resting
membrane is more permeable to K+ than to Na+ or other ions because it contains more K+ channels than
Na+ channels. The flow of K+ through these channels makes the major contribution to the resting
membrane potential of -70 mV, which is close to the K+ equilibrium potential.
3- Cl- ions: There is a higher concentration outside the cell than inside. The diffusional gradient tends to
push chloride ions into the cell. But since the cell at rest is negatively charged, it will tend to expel
negatively charged chloride ions, and so the electrical force pushes this ion toward the outside. These
forces are close to being in balance (they exactly balance at about 70 mV).

Action potential
When the neuron is inactive and polarized (inside negative; outside positive), it is said to be at its
resting potential. It remains this way until a stimulus comes along. The steps of an action potential are
described below:

1. Depolarization:
Graded potential. A graded potential is a change in the resting potential of the plasma
membrane in the response to a stimulus (# 1 in the picture below). A graded potential occurs
when the stimulus causes Na+ or K+ volted-gated channels to open. If Na+ channels open,
positive sodium ions enter, and the membrane depolarizes (inside of the cell becomes more
positive). If the stimulus opens K+ channels, then positive potassium ions exit across the
membrane and the membrane hyperpolarizes (inside of the cell becomes more negative). A
graded potential is a local event that does not travel far from its origin. Graded potentials occur
in cell bodies and dendrites. Light, heat, mechanical pressure, and chemicals, such as
neurotransmitters, are examples of stimuli that may generate a graded potential (depending
upon the neuron).

Action potential. Unlike a graded potential, an action potential is capable of traveling long
distances. If a depolarizing graded potential is sufficiently large, Na+ channels in the trigger zone
open. In response, Na+ ions on the outside of the membrane enter the cell and the membrane
becomes depolarized (as in a graded potential). If the stimulus is strong enoughthat is, if it is
above a certain threshold leveladditional Na+ gates open, increasing the flow of Na+ even
more. Na+ ions are driven into the cell due to both diffusional and electrical forces. This causes
complete depolarization of the neuron (from 70 to about +30 mV; the inside of the cell
becomes positively charged and the outside negatively charged) and an action potential is
created (# 2 in the figure below). In this state, the neuron continues to open Na+ channels all
along the membrane. When this occurs, it's an all-or-nothing phenomenon meaning once the
threshold is crossed, there's no turning back: Complete depolarization occurs and the stimulus
is transmitted along the axon. Increasing the intensity of the stimulus does not increase the

level of depolarization occurring during an action potential but it increases the number of
successive action potentials (# 3 in the figure below).

The opening of Na+ channels is both explosive and self-regenerating. As the voltage increases (i.e., the
cell depolarizes), more sodium channels open. As more sodium channels open, the cell becomes more
depolarized, and so on. This process is stopped by two factors: The cell reaches a point where the
diffusional and electrical forces on sodium balance themselves out, and so the opening of more
channels produces no further change. Also, at about this time, the sodium channels close and become
inactive.
2. Re-polarization:
The influx of Na+ in the cells causes another class of voltage-gated channels on the inside of the
membrane and specific to potassium ions to open allowing the K+ to move to the outside of the cell.
The move is driven by both the diffusional and electrical forces; with K+ moving toward its
concentration gradient and toward the outside which is now negatively charged. With K+ moving to the
outside, the membrane's repolarization restores electrical balance, although it's opposite of the initial
polarized membrane in that the Na+ are now on the inside and K+ are on the outside.
3. Hyperpolarization:
When the K+ gates finally close, the neuron has slightly more K+ on the outside than it has Na+ on the
inside. This causes the membrane potential to drop slightly lower than the resting potential, and the
membrane is said to be hyperpolarized because it has a greater negative potential. Hyperpolarization
can also be caused by influx of Cl through Cl channels. Note that influx of cations, e.g. Na+ through Na+
channels or Ca2+ through Ca2+ channels, inhibits hyperpolarization.
4. Refractory period:
The refractory period is the period when the Na+ and K+ ions are returned to their original sides: Na+ on
the outside and K+ on the inside by the action of the Na+/K+ - ATP-dependent pumps. While the neuron
is busy returning everything to normal, it doesn't respond to any incoming stimuli; the sodium channels
remain inactive. After the return of the ions to their rightful side of the neuron's cell membrane, the
neuron is back to its resting potential polarized state at which point the neuron is once again ready to
transmit another action potential.

Transmission of the impulse along the axon A closer view:

An action potential propagates along an axon,
from the axon hillock towards the synaptic
terminals, by being recreated in each adjoining
segment of the plasma membrane: one segment
causing the segment next to it to reach threshold.
The incoming Na+ ions spread through the cytosol in both
directions under the plasma membrane. The next
downstream segment of membrane, right next to segment
that just had an action potential, therefore become
depolarized to threshold, and initiates its own action
potential. The inrush of sodium ions, that this new action
potential caused, triggers the next segment of the axon to
initiation an action potential of its own, and so on down the
line.
The action potential travels in only one direction because
once a segment has initiated an action potential, it needs a
period of time, the refractory period, to recover. Until then,
it cannot initiate another action potential. Therefore,
despite the Na+ ions flowing both upstream and
downstream in the cytolsol, only the segment downstream
can be triggered to initiate another action potential. The
wave of action potential then travels from the axon hillock
to the presynaptic terminal of the axon.

Saltatory propagation
The speed at which an action potential travels along an axon is increased by increasing the size of the
axon. However, for myelinated axons, the myelin sheath provides insulation for the neuron and
increases the speed of the action potential transmission. The region of the axon covered in the sheath
does not have any channels present in the membrane, they are instead localised to the nodes of
Ranvier between the sheaths. In this way the potential can jump to each node with practically no loss
of signal. This type of transmission is called saltatory conduction.
Because the cytoplasm of the axon is electrically conductive, and because the myelin inhibits charge
leakage through the membrane, depolarization at one node of Ranvier is sufficient to elevate the
voltage at a neighboring node. Thus, the voltage at the first node of Ranvier extends spatially to the
next node of Ranvier. At each successive node, the membrane potential of the axon is thereby brought
to the threshold potential and initiates an action potential. Therefore, in myelinated axons, action
potentials do not propagate continuously as waves, but instead "hop" from nodes to successive nodes
along the axon. By doing so, the impulse travels faster than it would without the myelin sheets.

Chemical transmission of a nerve impulse at the synapse The arrival of the nerve impulse at the
presynaptic terminal stimulates the release of neurotransmitter into the synaptic gap. The binding of
the neurotransmitter to receptors on the postsynaptic membrane stimulates the regeneration of the
action potential in the postsynaptic neuron.

A gap called a synapse or synaptic cleft separates the axon of one neuron and the dendrites of the next
neuron. Neurons don't touch. The signal must traverse the synapse to continue on its path through the
nervous system. Impulses are carried across synapses as the following chemical changes occur:
a) Calcium gates open.
At the end of the axon from which the impulse is coming, the membrane depolarizes, voltage-gated
calcium channels open, and calcium ions (Ca2+) are allowed to enter the cell.
b) Releasing of neurotransmitters.
Calcium promotes the fusion of synaptic vesicles with the presynaptic membrane. Vesicles release
neurotransmitters into the synaptic cleft via exocytosis.
c) The neurotransmitter binds with receptors on the post-synaptic membrane (neuron or muscle).
The chemical that serves as the neurotransmitter moves across the synapse and binds to proteins
receptors on the post-synaptic membrane that's about to receive the impulse. Note that there are
different protein receptors for different neurotransmitters.
d) Excitation or inhibition of the membrane occurs.
Whether excitation or inhibition occurs depends on what chemical served as the neurotransmitter and
the result that it has. For example, in the case of Excitatory postsynaptic potentials (EPSP), the
neurotransmitter causes the Na+ channels to open, the neuron membrane becomes depolarized, and
the impulse is carried through that neuron. In cases of Inhibitory postsynaptic potentials (IPSP), the K+
or Cl- channels open (allowing movement of K+ outside the cell and Cl- inside the cell), the neuron
membrane becomes hyperpolarized, bringing the membrane further away from the threshold and
inhibition occurs.
Both IPSPs and EPSPs are tiny (less than 1 mV) and by themselves cannot trigger or stop an action
potential. When the many tiny IPSPs and EPSPs are added together, the postsynaptic cell uses the sum
of all these to decide whether or not to fire an action potential of its own. This decision point is called
the axon hillock or initial segment. In general, the dendrites and cell body of a nerve cell do not have
the machinery necessary to trigger an action potential. The junction between the cell body and the
axon is the first place this machinery exists, so it is the decision point. This arrangement also ensures
that the action potential moves away from the cell body, and toward the far end of the axon, where
the neuron makes contact with another cell.

After the neurotransmitter produces its effect, whether it's excitation or inhibition, the receptor
releases it and the neurotransmitter goes back into the synapse. In the synapse, the cell "recycles" the
degraded neurotransmitter. The chemicals go back into the membrane so that during the next impulse,
when the synaptic vesicles bind to the membrane, the complete neurotransmitters can again be
released.
Classes of Neurotransmitters and examples:
Acetylcholine
Biogenic amines (contains one or more amine groups)
Dopamine, Norepinephrine, and epinephrine
Serotonin
Histamine
Amino acids
GABA (gamma-aminobutyric acid)
Glutamate
Aspartate
Cysteine
Glycine
Peptides
Endorphins and encephalins
Somatostatin