SYNAPTIC TRANSMISSION: BLOCK 1

I.

Lecture 1: Neurophysiology/Synaptic Specialization

a. Basic Neurophysiology
i. Synaptic transmission is the communication between two nerve cells, while
neurophysiology is how the nervous system functions, or how individual nerve
cells function
ii. By controlling the distribution and movement of ions across cell membranes, our
cells can become excitable (i.e. generate electric current)
iii. Major ions in neural signaling (amounts below in mM):
1. K+
125 IC
5 EC
2. Na+ 10 IC
122 EC
3. Ca++ 0.0001 IC 2 EC
4. Cl5 IC
125 EC
5. A130 IC
0 EC
iv. Capacitance is the ability of something to store charge (think coulombs per volt)
The plasma membrane is a good capacitor; the larger the membrane area, the
larger the capacitance (neuron typically 1uF/cm2) capacitance is used to
estimate the size of a neuron
1. However, it is a leaky capacitor, so some ions can flow through (e.g. K+
can leak through even at resting conditions)
v. Resting membrane potential:
1. Equilibrium is determined by a combination of the concentration
gradient drive to be equal on both sides and the electrical forces acting on
an ion
2. At equilibrium, the # ions in = # ions out
a. The change in the electrical gradient across the membrane is larger
than the change in the concentration gradient a small number of
charges need to build up on the inside of the membrane to counter
a large concentration gradient (i.e. one K+ ion moving out of the
cell will have a greater effect on the electrical gradient than the
concentration gradient)
b. There is NO significant change in the concentration of ions on
each side of the membrane!
3. The equilibrium point for each ion is dependent on the distribution of
charges across a membrane; the EP equals the electrical potential (e.g.
Eqk) that can exactly balance the concentration gradient push of that ion
4. Eq = Equilibrium potential = ionic equilibrium potential = electrical
potential (at Eq no net movement of that particular ion!)
a. K+
-80 mV
b. Na+ +60 mV
c. Ca++ +120 mV
5. The two main factors that control the resting membrane potential are the
membranes resting permeability it has to ions, and the equilibrium
potentials of these ions (which results from IC/EC ionic distributions)

II.

6. The resting membrane voltage is -65 mV, closer to EqK, because the
membrane is much more permeable to potassium than sodium (-10 mV if
they were equally permeable)
a. Means that neither ion is at equilibrium, and thus, both have a net
flux out or into the cell, respectively
b. This flow of ions would eventually lead to the dissipation of the
unequal ionic distribution across the membrane
c. The sodium-potassium pump prevents this loss of ability to create
a resting membrane potential
i. Net outward flux of positive charge electrogenic pump
ii. Alters the balance of Na+ influx and K+ efflux; more
electrical driving force for Na+ to enter, and less electrical
driving force to move K+ out (closer to EqK, further from
EqNa)
iii. Net passive flux through leak channels = active pumping of
ions
Lecture 2: Driving Force and Ion Channels
a. Driving Force
i. The driving force is the push on an ion
ii. DF = Vm - Eqion
1. Negative = inward push of ions
2. Positive = outward push of ions
iii. Conductance, in biological terms, is equal to the flux of ions across the
membrane through protein pores or ion channels
1. Ability of charge to move from one point to another
2. Unit: Siemens (S)
3. Size of conductance depends on the number of charged particles moving
and how fast they move
iv. Resistance refers to how well a cell prevents the flux of ions (i.e. its leakiness)
1. The inverse of conductance (R = 1/g)
2. Unit: Ohms
3. The relative inability of electrical charge to migrate
4. The smaller the cell, the higher the resistance
v. Ohms Law: V = IR
vi.
b. Ion Channels
i. Ion channels are biological units that explain electrical resistance and
conductance. Protein pores that are selective for individual ions are called ion
channels.
ii. Ion channels are often gated by external influence:
1. Voltage potential difference across membrane (VG Na channels)

III.

2. Ligand binding to external surface (ionotropic glutamate receptors)

3. Ligand binding to internal surface (Ca-activated K+ channels)
4. Membrane Stretch (mechanically gated channels)
c. Voltage-Gated Ion Channels
i. Structure inferred from biochemical and molecular biological studies of the amino
acids that make up the proteins
ii. Basic structure of all voltage-gated ion channels: 4 domains (groups of
transmembrane regions) that each contain 6 alpha-helical membrane spanning
segments
1. Pores formed from the 4 domains
2. P-loop the extracellular loop that dips back into the channel between the
5th and 6th transmembrane segments
3. S4 helix voltage sensor (positively charged amino acid every third
position)
iii. For Na+ and Ca++ channels, the 4 domains are part of one protein; they are
separate proteins for K+ channels
iv. Ion selectivity is produced by narrowing of the pore at selective binding sites for
an ion; an ion shuffles through the pore by briefly interacting with these binding
sites
d. Channel Function
i. Voltage-Dependence of Activation Curve
1. Indicates if there is a pathway open
a. Channels dont open unless all 4 domains S4 segments move
2. Given in fractional activation or normalized conductance
3. You must consider driving force plot with voltage-dependence of
activation plots to actually see if an ion will move across the membrane
ii. Driving Force Curve
1. Relates membrane potential to driving force on an ion
iii. Current-Voltage Relationship Curves
1. Relationship between the amount of current that flows at any given
voltage
Lecture 3: The Action Potential
a. Action Potentials
i. Action potentials are a mechanism for neurons to carry out long distance signaling
ii. During an action potential, there is a transient increase in the permeability of Na+
to the membrane.
iii. Three things set up the neuron so that it can use an action potential as a signal:
1. Distribution of ions across a membrane
2. Resting permeability to K+
3. Resting potential
iv. Depolarization caused by influx of positive ions
1. This increases the probability that voltage-sensitive Na+ channels will
open
2. As Na+ channels open, more positive ions flow in, and thus, cause even
more Na+ channels to open, eventually reaching threshold
v. Threshold
1. Subthreshold stimulus = depolarizations that are not big enough to
significantly activate sodium channels

2. Depolarization Activation of Na channels increased

influx/conductance of sodium Activation of more Na+ channels
conductance of more sodium, etc.
a. This explosive depolarization is the action potential
3. Typical membrane voltage at peak of AP is about +30 mV
4. At the peak of an action potential:
a. Membrane is more permeable to Na+ than K+
b. Some voltage-gated K+ channels begin to open, so Vm does NOT
equal Eq-Na+
5. Falling Phase:
a. Repolarization mainly controlled by the increase in the
permeability to K+ through voltage-gated K channels
i. Delayed increase in K+ conductance pulls Vm to Eq-K
1. Often causing an after hyperpolarization (AHP)
b. Even without the opening of VG-K+ channels, the cell could
repolarize (slowly) through the inactivation of VG-Na+ channels
i. Channels close in the continued presence of the activating
stimulus (voltage)
ii. Two potential mechanisms of channel inactivation:
1. Delayed voltage-activated pinching of the gate
closed again (c-type)
2. Ball and chain plugging of an open channel pore
a. Fast; type used for sodium channels
b. Contributes to repolarization and refractory
period
c. Action potentials do NOT result in significant changes in ion
concentrations across the membrane
i. Remember, ionic charges required to change membrane
polarization are insignificant with reference to the
concentration gradients in the large cell volume
ii. AP is produced because the Na+ and K+ fluxes charge the
membrane, NOT because the fluxes change the ionic
distribution across the membrane
vi. KCl Injection
1. Vm of cardiac muscles cells is near -65 mV, and because of the
distribution of K+ across the cell membrane, that creates an Eq-K that is
near -80 mV
a. Get rid of unequal K+ distribution (hyperkalemic solution) EqK changes to near 0 mV No negative resting potential = Na
channels are inactivated
b. Inside of the cell needs to return to a negative potential between
impulses to allow Na+ channels to recover from inactivation so
they can open and depolarize a cell again
vii. Action Potential Propagation Over Great Distances
1. Plasma membrane = leaky insulator
a. A small depolarization caused by injected current would slowly
decay in size as it travels over distance

IV.

b. Action potential occurs in one area of a neuron (i.e.

suprathreshold) depolarizes the neighboring membrane area
above threshold regenerating wave of depolarization (large
enough to create the explosive positive feedback DP = action
potential) AP sweeps down nerve in both directions
i. To do this in every segment, it is SLOW
ii. Characteristics that allow this to spread quicker:
1. Concentrated Na+ channels in patches
2. Myelination allows the AP to spread without
much decay between Na+ channel patches (i.e.
Saltatory Conduction)
3. Diameter bigger diameter = less axial resistance
b. Microelectrode Recording
i. Set-Up: Two electrodes filled with concentrated salt solutions placed on either
side of the membrane (draw set-up); one electrode inside, one outside allows you
to record the resting membrane potential; another electrode will then be inserted
into the cell for purposes of current injection
ii. Directly records the potential difference between them by using an amplifier and
volt meter (oscilloscope); does NOT show you the charges that flow across the
membrane
iii. For large, leaky cells (small resistance), smaller voltage change produced by
current injection than for small, tight cells (higher resistance)
1. Ohms Law: V = IR
c. Voltage Clamp
i. Used to measure the flux of charges across the membrane (i.e. current), e.g. the
flux of Na+ during an action potential
ii. Obstacles of measuring flux:
1. Current flow change in membrane potential change in driving force
for each ion change in ionic flux
2. Currents can flow across the membrane in both directions (e.g. Na+ in, K+
out) interpretation of overall current can be difficult
iii. Solutions to allow for flux measurement:
1. Vm can be held steady or clamped while measuring the magnitude and
time-course of current flux (command potential)
2. Selective measurement of one ions current flux
a. Block certain ion channels
b. Alter ionic environment
iv. Example: command potential changed from -70 mV to -15 mV
1. Amplifier delivers positive current to the inside of the neuron to drive it to
-15 mV Na+ channels open in response; Na+ influx Negative
current applied to exactly balance Na+ current influx (and thus, prevents
depolarization)
2. The voltage clamp provides an exact measure of the negative current
required to hold Vm at command potential; this is an exact mirror image
of the Na+ current in response to the command potential
Lecture 4: Patch Clamp Techniques and the History of Synaptic Transmission
a. Patch Clamp

i. A variant of the voltage clamp that is easier to perform on smaller cells and allows
the direct observation of ionic current flux through a SINGLE ion channel
ii. Glass pipette with a tiny opening is used to make contact with a patch of the
neuronal membrane; suction applied to seal pipette and membrane
iii. Currents flowing through single channels are called microscopic currents; these
current measurements look like steps; macroscopic currents represent the sum
of the microscopic currents, and the likelihood a channel will be open at a
given time during depolarization of the membrane

b.

c.

d.

e.

iv. Cell-Attached Recording

1. Single channels are isolated on cell surface, so you can study the effects of
various manipulations on the function of individual channel proteins
a. Info about the behavior of each channel protein adds to create the
total current flux measured using voltage clamp
v. Whole-Cell Recording
1. Strong suction disrupts membrane, and interior pipette becomes
continuous with the cell cytoplasm
2. This arrangement allows measurements of electrical potentials and
currents from the entire cell
vi. Inside-Out Recording
1. Mild suction allows pipette to pull away a small vesicle of membrane;
vesicle exposed to air, creating an inside-out patch of membrane
2. Allows change of the medium that the intracellular surface is exposed to
vii. Outside-Out Recording
1. Pulling off pipette during whole-cell recording; outside of membrane
fragment re-seals
2. Good for studying the effects of extracellular signals such as
neurotransmitters
Charles Sherrington
i. Introduced concept of the synapse
ii. Was studying spinal cord reflexes stimulating sensory nerve and recording from
motor nerves and hypothesized that at the point of contact between a sensory
and motor neuron, there was a unidirectional info transfer junction
Galvani and DuBois-Reymond
i. Experimental set-up
1. Connected iron rod to frog spinal cord
2. Measured contractions of frog leg muscles; also measured flow of current
in both the nerve cell and muscle cell when excited by the nerve
3. Believed (incorrectly) that this animal electricity flows directly from
nerve to muscle, in the brain, between neurons
Camillo Golgi and Reticular Theory
i. Golgi developed stain for individual neurons that showed complete processes
associated with individual neurons
ii. He (incorrectly) interpreted these as direct connections between cells
Ramon y Cajal
i. Used Golgi Stain to argue that axons and dendrites are NOT continuous with
one another
ii. Believed that axons ended in a certain apparatus (which would later be named
the synapse by Sherrington)

f. Sir Henry Dale

i. Added ACh derivatives to the heart, resulting in slowing of heart rate, mimicking
the action of the vagus nerve on the heart
ii. Based on results, argued that synaptic transmission was chemical; however, others
claimed it was just a synthetic compound mimicking the brain
g. Otto Loewi
i. Model: Two frog hearts were placed in separate baths, connected by a pump. One
heart was innervated by the vagus nerve, the other was not.
1. Would not have worked with many other animals, such as dogs; one,
because of the high concentration of AChE, and two, ACh breaks down
faster at warmer temperatures (problem for warm blooded animals)
ii. When pump was off (aka solution 1 couldnt flow to solution 2) and the vagus
nerve was stimulated, the firsts heart rate slowed, while the seconds remained
constant. When the pump was on, connecting the solutions, and therefore, their
chemicals, both heart 1 and 2 had slower heart rates following vagus nerve
stimulation
iii. Experiment allowed pharmacologists to conclude that information was
transferred at synapsis via a chemical stimulant
h. Electrical Synapses
i. Usually between large presynaptic cell and small postsynaptic cell
1. Need a postsynaptic cell with larger resistance in order to produced a
voltage that is large enough to depolarize for a given current
ii. First seen in a motor synapse in the crayfish; axo-axonic
iii. Evidence for electrical synapses
1. Latency between pre and post AP is almost non-existent
2. Subthreshold activity in the presynaptic cell creates a graded potential in
both the pre and post cell; presynaptic voltage change is proportional (not
identical) to postsynaptic voltage change
iv. Purpose of Electrical Synapses
1. Fast transmission for speed
2. Synchrony; linking of activity between neurons
a. e.g. Stereotypic behaviors like ink gland discharge in sea slugs;
once first neuron fires, all neurons fire together
b. Hebbian synaptic changes the cells that fire together, wire
together important for development
3. Exchange of chemical messengers though the channels
a. Large channel pores are molecular basis for gap junctions; nonselective for conductance
i. Smaller synaptic cleft
ii. Large conductance
iii. All inorganic ions, and some organic compounds such as
small peptides, cyclic nucleotides (cAMP), IP3
iv. Experimental markers suggest the above
v. Formed by a pair of hemi channels (connexons) in preand post- membrane (with 4 TMR/subunit)
vi. Channels are typically open; can close with lowered pH or
elevated intracellular Ca++; rectifying synapses are
unidirectional because they are voltage sensitive;
phosphorylation can also alter conductance

V.

v. Electrical Synapses in Humans

1. Developing cortex
a. Groups of postsynaptic neurons that will eventually all receive
similar synapses in adulthood
2. Developing spinal cord
a. Allows synapse formation while electrical synapses are present,
and then triggers competition that leads to synapse elimination
when electrical synapses disappear
3. Retina (adults too)
a. Likely to help us detect the direction of a moving light stimulus by
taking advantage of the synchronous activity of electrically
coupled ganglion and amacrine cells
i. Advantages vs. Costs of Chemical Synapses
i. Advantages
1. Plasticity modulation and changes in synaptic transmission underlie
all behavior
2. Excitation/inhibition is easy
3. Can excite a LARGE postsynaptic cell
ii. Disadvantages
1. Slower
2. More energy; less efficient
3. Not as hard-wired and can be disrupted
Lectures 5 and 6: Synapses and Quantal Theory
a. Fatt and Katz
i. In 1950, record the postsynaptic electrical event in a muscle cell
1. AP in pre cell is about 1 msec before the AP in post cell synaptic delay
2. The synaptic potential in the post synaptic cell is termed the end plate
potential, and is only seen when recorded near the endplate
a. To observe an EPP without the complication of an AP, you can
selectively block sodium channels
ii. In 1952, they observed spontaneous randomly occurring deflections (about 0.5 to
1 mV in size), which they termed miniature end plate potentials
1. They had many of the same qualities of EPPs, including the same:
a. Time course
b. Effects on ACh receptor blockers
c. AChE blockers (increase and elongate both)
d. Nerve degeneration after cut
2. Also, only observed when recording near the endplate, suggesting a local
release of transmitter near the nerve terminal, but in the absence of an AP
in the presynaptic neuron
3. Tested their theory by applying various amounts of ACh to endplate, and
recording responses.
a. Results showed graded responses that were much smaller than
mEPPs, suggesting that mEPPs were the result of a synchronous
release of a large number of ACh molecules that remain in the
synaptic cleft for a transient period of time
b. Later, Katz and Miledi used patch clamp techniques to measure
flux through a single ACh receptor channel, and estimated that

about 2000 receptors opened for each mEPP (approx. 5000 ACh
molecules per mEPP)
b. Quantal Nature of Synaptic Transmission
i. Do lots of mEPPs make up one EPP?
1. Katz varied the extracellular ion concentrations, and realized that action
potential-evoked transmitter release required extracellular Ca++
2. When EC Ca++ was dropped to very low levels, the EPP size got smaller
some even nearly as small as mEPPs
3. The above approach was used to accurately measure the size of EPPs and
mEPPs
a. When an EPP occurred, the amplitude was a multiple of the mEPP
amplitude (i.e. EPPs fluctuate in amplitude by the size of mEPP)
b. As the calcium concentration outside was increased, the mEPP size
did not change (at one particular voltage), but the EPP size did
increase
i. Note that the EPP represents a non-linear summation of
quanta as the membrane potential increases, the
smaller the amount is that each quanta contributes to
change in an EPP!! This is due to the change in
membrane potential resulting in a change of driving force!
c. Calcium increases the likelihood that transmitter will be released
ii. The Quantal Theory of Synaptic Transmission
1. Transmitter is only released as a fixed number of molecules (quanta)
a. Occurs via a membrane bound synaptic vesicle that contains
thousands of molecules of transmitter
i. NMJ: 150 quanta per AP within 1 msec
ii. CNS: 1-10 quanta per AP
b. ACh binds to receptor channels, which have equal conductance for
Na+ (in) and K+ (out), and thus, have an Eq = -10 mV
i. The closer Vm is to -10 mV, the less driving force there on
ions to pass through these channels; this results in smaller
mEPPs at higher membrane potentials
2. Occasionally, quanta are liberated spontaneously (e.g. mEPPs)
3. A presynaptic action potential increases the probability that these
quanta will be released (by causing Ca++ to enter the nerve terminal)
iii. Quantal Analysis
1. Assuming that a single vesicle is equal to a mEPP, and that an AP induces
synchronous fusion of many vesicles to result in an EPP, we can try to
understand how strong synapses are, and how strength changes with
synaptic events
2. Quantal analysis determines if a change in strength of communication
between neurons is due to a change on the presynaptic or postsynaptic side
of the synapse
3. If you can measure the size of mEPPs and EPPs, you can divide the mean
EPP/mean mEPP and get the quantal content, m, i.e. the number of
effective quanta/vesicles released in response to a nerve impulse
4. The mean postsynaptic response amplitude can be determined by m =
npq, where:
a. n = number of release sites

b. p = probability of release from each site

c. q = synaptic response to release of NT from a single vesicle
5. Changes inform you of the mechanism of action of a drug/alteration:
a. Change in n = change in the number of presynaptic release sites
b. Change in p = change in the prob. that synapse releases transmitter
c. Change in q = change in postsynaptic sensitivity to transmitter
6. Assumptions:
a. If drugs are applied, time will be needed to see a change in n,
number of release sites
b. The probability of release, p, is the same at each release site
c. The quantal size, q, is a measure of our ability to measure the
postsynaptic response to a single package
7. In order to use quantal analysis to determine the mechanism of action, you
need to be able to determine the above variables
a. Such can be done using a voltage clamp to measure mEPPs and
EPPs (that way Vm remains constant and doesnt alter the driving
force, and thus, the mEPP); use the current measurements to
determine the size of mEPPs and EPPs
b. Change in EPP without change in mEPP OR change in frequency
of mEPPs = change in P
c. Change in EPP and in mEPP = change in q
8. Problems with quantal analysis in the CNS
a. Many synaptic sites on single CNS neuron
b. All release sites may not have the same probability, p
c. Difficult to detect mEPSPs and EPSPs above background noise
d. Number of postsynaptic receptors can be very small (and thus,
saturate quickly, making it difficult to measure the magnitude of
release)