Neurophysiology – lecture 23

April 7, 2011

1 Review:
1. General electrical model to explain Electrical Synaptic Transmission:
(a) For A.P. propagation along an axon we could describe the axon as made up of a series of segments
each of which contained the channels needed for A.P. generation and each segment was separate
from adjacent segments by a moderately sized longitudinal resistance, ri . In this model sufficient
current passes from a cell experiencing an A.P. through ri to depolarize the membrane capacitor
of the adjacent segment by a sufficient amount that an A.P. was triggered in the adjacent segment.
(b) On the other hand an A.P. generated in one neuron does not depolarize an adjacent neuron because
there is the substantial resistance of 2 membranes between them and there is a relatively low
resistance current pathway through the narrow channel filled with extracellular fluid separating
the 2 neurons. In this case most of the current generated by an A.P. in one neuron flows through
the relatively low resistance current pathway provided by the extracellular fluid between the
neurons and almost none of it flows through the membrane of the second neuron. So no response
is produced in the adjacent neuron when an action potential is produced in the other neuron.
(c) In the case of Electrical Synapses there are specialized structures called “gap junctions” which
provide a somewhat higher resistance pathway for current flow than cytoplasm, but a much lower
resistance pathway than the membrane of an adjacent neuron would provide. Thus, sufficient
current does flow through the gap junction to produce some depolarization in the postsynaptic
neuron.
(d) The depolarization or hyperpolarization in the postsynaptic neuron (∆V mpost ) is always less than
that produced in the presynaptic neuron (∆V mpre ).
∆V mpost
The “coupling ratio” between the neurons =
∆V mpre
2. Functional uses of Electrical Synapses
(a) Along the Earthworm Giant Fiber system electrical synapses between giant axons in successive
segments of the worm produces rapid A.P. propagation along the multiple axon system.
(b) At the synapse between the giant fiber of the crayfish ventral nerve cord and the motor axons
which innervate its muscle fibers an electrical synapse results in fast synaptic transmission (0.1ms)
and thereby fast escape responses.
3. The many electrical synapses between the muscle fibers in the heart result in excitation produced in
one spot to rapidly involve all the muscle fibers in a given area of the heart (e.g., the ventricles),
thereby leading to a synchronous contraction of the ventricular muscle (a heart beat).

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2 Chemical Synapses
1. Structure of Chemical Synapses.
(a) It was presumed that students in neuroscience already know the three main features which charac-
terize chemical synspses:1) Synaptic vesicles, 2) a Synaptic Cleft, and 3) a Postsynaptic Density.
(See Figure 16A.)
2. The sequence of events occurring during the course of chemical synaptic transmission:
An A.P. propagated into the presynaptic terminal →
An influx of Ca2+ into the presynaptic terminal →
Synaptic vesicles fuse to the presynaptic terminal membrane →
The fused vesicle releases its contents into the synaptic cleft →
Some of the released neurotransmitter diffuses across the cleft →
Some of the transmitter binds to postsynaptic receptors →
Ion channels open in the postsynaptic membrane →
A postsynaptic response occurs.
In this course we will deal only with the first 2 and last 3 steps in this sequence of events.

3 The neuromuscular junction - the most studied synapse

1. Structure: For each muscle fiber there is but one axon and one synapse. (See Figure 16B)
On the other hand a particular motor neuron axon can innervate many muscle fibers.
The NMJ is very large for a synapse; it actually functions as about 300 synapses acting simulta-
neously.
2. Advantages of the NMJ
(a) Since there is only 1 synapse/muscle fiber the source of the postsynaptic potentials is clear.
(b) It is easy to stimulate the entire ventral root or motor nerve to elicit a postsynaptic response, the
action potentials generated in other axons than the axon innervating a particular muscle fiber do
not effect the recordings.
(c) Muscle fibers are big and easy to record from.
(d) The locus of the presynaptic axon on the surface of the muscle fiber is often visible.
3. Problems with the NMJ
(a) Because it appears like the summation of 300 synaptic responses the postsynaptic response is a
large depolarization (about 50mV) and generally triggers an action potential in the muscle fiber.
Muscle action potentials cause the muscle fiber to contract which pulls the intracellular recording
electrode out of the muscle fiber and abruptly ends the recording session.
(b) To prevent such abrupt endings of recordings a low concentration of curare is added to the bath
containing the nerve muscle preparation. The curare reduces the amplitude of the postsynaptic
response so an action potential is not elicited. by each postsynaptic response.

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4 What couples presynaptic action potentials to neurotransmitter
release?
1. This could be rephrased, “What causes a presynaptic A.P. to release neurotransmitter?”
2. There are 3 ways to answer any causal question. They include:
ADDITION of the presumed “causal agent”:
(a) If Ca2+ is added to a medium bathing a nerve - muscle preparation, the amplitude of the postsy-
naptic response elicited by stimulation of the axon innervating the muscle fiber increases markedly.
(See Figure 16C.)
(b) Katz and Miledi, 1951 and 1967, discovered that if Ca2+ was ionophoretically applied to an NMJ
immediately before it was depolarized that a postsynaptic response would be recorded even though
there was no Ca2+ elsewhere in the bathing solution (See Figure 16D, part B).
The postsynaptic response will be elicited only if the Ca2+ is applied just before or during the
depolarization; it will not be elicited if the Ca2+ is applied after the depolarization.
This suggests that Ca2+ must come from outside the synaptic terminal to elicit neurotransmitter
release. The need for the depolarization suggests that channels are being opened to allow Ca2+
to flow into the presynaptic terminal.
(c) In the 1990’s “caged” Ca2+ compounds, like DM-Nitrophen, were developed which bind Ca2+
tightly but then release it rapidly when these compounds are exposed to UV light. In the squid
giant synapse “caged” Ca2+ compounds can be injected into the presynaptic terminal. A flash of
UV light onto the terminal will then produce a postsynaptic response.
(d) Collectively, these results indicate that a sudden influx of Ca2+ coming from outside the presy-
naptic terminal raises the intra-terminal Ca2+ concentration and this increase in intra-terminal
Ca2+ concentration causes a release of neurotransmitter.
SUBTRACTION of the presumed “causal agent”
(a) In Figure 16B in the bottom set of traces (labeled “c”) it is shown that if Ca2+ is applied shortly
before the depolarizing stimulus pulse that a postsynaptic response is elicited in the muscle fiber.
However, if Mg2+ is previously applied and still present at the synapse, the postsynaptic response
does not occur.
Mg2+ blocks the pathway (channels) that Ca2+ uses to enter the presynaptic terminal, so that in
the presence of sufficient Mg2+ there is no influx of Ca2+ into the presynaptic terminal. Thus,
Mg2+ effectively subtracts the intra-terminal increase in Ca2+ . Other divalent and trivalent
cations also block Ca2+ entry, such as Cd2+ , Mn2+ , La3+ , etc.
(b) Compounds which contain two carboxylic acid groups near one another bind divalent Ca2+ ions
and remove them from being “free” in the cytoplasm. This is referred to as calcium “chelation.”
BAPTA is such a chelater which binds Ca2+ very rapidly. When BAPTA is injected into the
presynaptic terminal of the squid giant synapse, presynaptic action potentials no longer elicit
postsynaptic responses.
(c) Thus removing the ability of the Ca2+ concentration inside an axon terminal to increase removes
the ability of action potentials invading that terminal to produce postsynaptic responses.