BRAIN SCIENCE

with

Ginger Campbell, MD

Episode #210

Neuroscience Basics with Focus on Neurotransmitters

Aired July 28, 2023

[Music]

INTRODUCTION

Welcome to Brain Science, the podcast that explores how recent discoveries in
neuroscience are unraveling the mystery of how our brain makes us human. This is
episode 210, and I'm your host, Dr. Ginger Campbell.

If you're new to Brain Science, I want to encourage you to jump right in and don't be
discouraged if you don't get everything on the first listen. My goal is to make
neuroscience accessible to everyone because I strongly believe that understanding
how our brain really works is essential to being a citizen in the 21st century.

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This month's episode is a little unusual because for the last 15 years or so, I have
focused on interviewing neuroscientists, and exploring how their work is important
to understanding ourselves and each other. But today, I'm reviewing an important
but complex topic: neurotransmitters.

 Even if you've never heard this word before, you have no doubt heard about
molecules like dopamine. Unfortunately, most media coverage is superficial and
often inaccurate, which is one reason this episode is long overdue.

I last talked about neurotransmitters back in episode 8, which was released in 2007.
The basic principles remain the same, but the discoveries in this area have really
exploded. The main reason I'm devoting this month's episode to neurotransmitters is
that next month, molecular biologist Seth Grant will be returning for his record sixth
time.

He is incredibly good at making his work understandable, but I hope this extra
background information will be helpful. Obviously, this material is going to be
completely new to some of you and just review for others.

Please listen all the way to the end of the episode because I will review the key ideas
and share a few brief announcements, including an update on my move to New
Zealand.

[Music]

REVIEW

Okay, let's jump into this month's episode. I'm going to start with a few basics.
Neurons and glial cells are the key cells of the nervous system. Neurons are
distinguished by their ability to generate action potentials, which are all or nothing
electrical spikes that spread from neuron to neuron.

The outgoing fiber from the neuron is called the axon, and the place where the little
extensions that receive axon signals are called dendrites. Molecular biology and
genomics are revolutionizing our understanding of how the brain works. That's one
reason why I enjoy so much talking with Seth Grant who will be back next month to
tell us more about what's going on inside the synapse.

Anyway, the Human Genome Project has determined that the human genome only
has about 20,000 genes that code for proteins. This was actually a surprise. They
were expecting the number to be more like 100,000.

The nervous system expresses about 14,000 of these genes, and there are 6,000
proteins that appear to be unique to neurons. The Allen Brain Institute has
determined that different brain areas actually express different proteins, that means
different genes are turned on.

And Seth Grant has determined that gene expression actually changes throughout
the lifespan. In fact, it's predictable like a calendar.

One reason molecular biology has become so important as neuroscience, is that

knowing which genes code for certain proteins allows molecular biologists to tag the
proteins in various ways, and they can also study what happens if a normal gene is
replaced with one with a mutation.

1 © Copyright 2023 Virginia A Campbell, MD

 Neurons can be classified in several different ways. Traditionally, they were classified
by morphology, which is what they look like under the microscope, but now, they're
classified by their genetic makeup, and they also are classified by which
neurotransmitter they release.

It used to be assumed that they could only release one type that's been proven to be
false, but they're usually identified by the main neurotransmitter that they release.

Now, the glial cells actually outnumber the neurons, although the exact ratio is
debated. They used to be seen just as scaffolding, but we now know they do much
more. They take up important molecules like neurotransmitters. They modulate
synaptic transmission, they actually release their own important substances.

Oligodendrocytes and Schwann cells are types of glial cells that actually make
myelin, which coats the axon to make transmission faster, and glial cells also appear
in the enteric nervous system, which I'll talk about shortly.

If you want to know more about glial cells, I refer you to episode 169, which is an
encore episode with Doug Fields about what glial cells do. I also want to briefly just
talk about brain structure starting from front to back. Again, there's more details
about this in episode 118, which is an interview with David Bainbridge, author of
Beyond the Zonules of Zinn, a really cool introduction to brain anatomy.

Anyway, there's the cortex, which is the covering of the brain. It covers the lobes of
the brain known as the frontal, parietal, temporal, and occipital lobes.

The temporal lobes, which are right on the sides of your heads above your ears, they
include auditory and olfactory cortex, and then structures in the middle or medial
structures like the hippocampus and the amygdala.

The amygdala is actually part of the temporal lobe, although it has a subcortical
structure. What this means is that it has nuclei or clusters of neurons instead of the
layering structure of the cortex.

There's another lobe called the insular lobe, which people don't usually think about
because it is the only region you can't see from the surface. It's located within the
lateral sulcus between the temporal, frontal, and parietal lobes, sort of where they
come together. And this is where interoception appears to be processed.

Association areas are really important, and the parietal cortex is known to have
association areas for integrating sensory information. The temporal cortex also has
association areas for auditory information and language processing.

Okay, so as we start to move back or down in the brain, we get to the subcortical
structures. The basal ganglia are very important to voluntary movement, and then
there's the brain stem, which is the area below the cortex before you get to the spinal
cord. It's essential for basic life functions like breathing.
It consists of the midbrain, which actually contains also the superior colliculus,
which is important in visual processing, and then the other structures are called the
pons and the medulla. Tucked underneath the occipital lobe is the cerebellum.

2 © Copyright 2023 Virginia A Campbell, MD

 Now, rather than to get too bogged down in basic neuroanatomy, I again, refer you
back to episode 118, and like I said, the book, Beyond the Zonules of Zinn is really
good, and also Frank Amthor’s book, Neuroscience for Dummies has a good
description of all the basic neuroanatomy ideas.

Instead, what I want to do today is to focus on neurotransmitters and the synapse.

My goal is to give you background for next month's interview with molecular
biologist, Seth Grant.

One key idea I want to share is that neurons are actually very different than the way
they're modeled in the field of artificial intelligence. In a sense, they're hybrid
because they have both analog and digital properties.

You could think of the action potential as a digital signal since it's basically an on/off
kind of thing. But at the synapse, there are chemical signals called neurotransmitters
that are released, and this is very much an analog process.

When the action potential reaches the synapse, a neurotransmitter is released by the
presynaptic neuron. Then the neurotransmitter interacts with the receptor on the
postsynaptic neuron, or if it's the neuromuscular juncture, a receptor on the muscle.

One key idea that I'm going to say many times today is that what happens next is
determined by the receptor, not by the specific neurotransmitter. This is why modern
neuropharmacology is trying to target specific receptors, rather than trying to block
or mimic the neurotransmitters.

To understand how neurotransmitters work, we need to know a little bit about the
structure of cell membranes. The cell membrane is the essential barrier between
inside the cell and the outside world. It's made up of something called a lipid bilayer.

Now, lipid molecules have two ends; a fatty hydrophobic end, and a so-called,
hydrophilic polar group that contains phosphate. The key idea here is that the polar
head has a slight electrical charge that causes it to be attracted to water, while the
fatty end repels water. That's why oil and water don't mix.

To picture this, just imagine that your fingers are fatty acids, and the knuckles of
your hand are the phosphate group. If you point your fingers toward each other, this
is the way the fatty acids line up to create the lipid bilayer that surrounds every cell.

So, this lipid bilayer is embedded with carbohydrates and proteins, with proteins
making up more than half of the actual cell membrane. Some proteins create
channels that allow ions and other small molecules to go in and out. That's why it's
considered a semi-permeable membrane.

There are only very small molecules that can just go through the membrane by
simple diffusion. This is gases like oxygen, carbon dioxide, and nitrous oxide, and
ethanol. Some of the ion channels allow passive diffusion down a concentration
gradient, but most of the movement requires what's called active transport, which
means it uses energy in the form of ATP.

3 © Copyright 2023 Virginia A Campbell, MD

 For example, the sodium-potassium pump, which is ubiquitous in all cells, pumps
three sodiums out and two potassium in to create a voltage difference that's known
as the resting potential of the cell. All the pumps work with a similar method.

Some are voltage-gated, which means they're open or closed based on voltage. The
action potential basically changes the voltage inside the cell from negative to positive.
For more on this, I refer you back to Neuroscience for Dummies and also, the
episode about The Spike with Mark Humphries. That was episode 186, and
Neuroscience for Dummies is 197.

So, now we're ready to talk about the types of receptors because the receptors are
where the action is. There are two types. One is called ionotropic or ligand-gated, and
the second is called metabotropic or G-protein-coupled, and now they're usually
called metabotropic G-protein-coupled altogether.

These names are tongue twisters, but the basic difference between them is really
pretty straightforward. The ionotropic receptors are actually ion channels that are
directly opened and closed by the ligand, which may be the neurotransmitter.

The metabotropic receptors use the so-called second messenger, which means that
they initiate a series of intracellular events. Just remember, ionotropic means that
ion channels are involved, and they act rapidly because it's a direct opening closing.
Whereas, the second messengers are involved with the metabotropic receptors, so
they act more slowly.

Since I first talked about these two types of receptors back in episode eight, much
more has been learned and this includes identifying many different receptors with
many different behaviors. Again, I refer you back to Neuroscience for Dummies for a
very basic discussion of this.

For example, glutamate is the main excitatory neurotransmitter in the central

nervous system, but it actually has its own families of receptors, including the
famous NMDA receptor that Frank Amthor and I talked about in episode 197.

The NMDA receptor is interesting both from a clinical and a basic science
perspective because it actually requires two molecules of glutamate and two
molecules of glycine because it has an additional voltage channel that is blocked by a
magnesium ion.

So, let's review briefly what happens during an action potential.

The resting potential inside the cell is usually around minus 65 millivolts. The
threshold of minus 55 millivolts opens the sodium channels and the sodium ions
rush in, and this produces the positive spike. It's digital because it always looks
exactly the same.

Later on, the potassium channels open and they stay open a while, which this allows
the cell to actually get more negative than its resting potential, and that's called the
refractory period because the action potential’s not going to happen during the
refractory period. So, there's a limit to how often any given cell can generate an
action potential.

4 © Copyright 2023 Virginia A Campbell, MD

 Then the action potentials are propagated along the axon of a neuron. They're not
propagated directly between neurons, and I'll talk more about this in a minute.

So, what happens at the synapse, that tiny gap between the axon of one neuron, and
the dendrite of the postsynaptic neuron?

First, it's interesting to note that while mammalian cells rely on chemical synapses,
the ones that are described in every textbook; invertebrates can also have electrical
synapses. And these occur at what's called a gap junction. They're very rapid, but the
postsynaptic responses leads to a much smaller signal, so they're not good for long
distance communication.

In contrast, chemical synapses are slower, but they're more adaptable. They allow the
signal to be boosted and even inverted, and they allow for the very important role of
second messengers. So, two types of chemical transmission occur that are
corresponding to the two types of receptors.

There's fast, direct transmission, involving the ionotropic receptors, and then there's
slow indirect transmission, which is involving the G-coupled-proteins and second
messengers, and this is actually neuromodulation.

Now, some neurotransmitters can do both depending on which type of receptor is

present. The key idea is that this multiplies the power of the primary
neurotransmitters and increases their flexibility.

I want to mention at this point, a great book called The War of the Soups and the
Sparks, which describes the discovery of the first neurotransmitter that was
discovered, which is acetylcholine. And you can get this book on KindleTM, I highly
recommend it.

So, let's talk a little bit about the categories of neurotransmitters. Acetylcholine was
the first one discovered, and acetylcholine is what's known as an amine. Then there
are others that are basically small neurotransmitters of different types.

Adenosine is what's known as a purine, the monoamines, which I'll talk about more
later, which includes histamine, dopamine, serotonin, epinephrine, norepinephrine,
amino acids, glutamate, aspartate, GABA, glycine, and finally, there are
neuropeptides. I'm going to talk about these different things as we go a little bit
further along.

But going back to acetylcholine, the criteria for what constitutes a neurotransmitter
is actually based on acetylcholine. So, here are the requirements.

For something to be considered a neurotransmitter, it has to be endogenously

produced or present in the pre-synaptic neuron. When stimulated, the presynaptic
neuron must release the chemical and it must produce a response in the postsynaptic
neuron. That response must be mimicked by exogenous application.

That means if you add that same molecule to the postsynaptic neuron, you should see
the same responses you got as if it was stimulated by the presynaptic neuron. And

5 © Copyright 2023 Virginia A Campbell, MD

 the chemical has to be endogenously removed at the synapse, and you'll understand
why that's important in a little while.

So, if all those requirements are met, then it's considered a conventional
neurotransmitter which is released from vesicles. And the conventional small
molecule neurotransmitters are synthesized by enzymes at the presynaptic terminal,
and that includes the compounds that I just mentioned.

There's also a second group that act as neurotransmitters, which are neuropeptides,
and they're a little bit different because they aren't made at the presynaptic terminal,
they're made in the cell body and also they tend to be released actually away from the
synapse and have more of a neuromodulatory and slower effect. But I'll talk more
about neuropeptides in a few minutes.

As I was saying, the criteria for a conventional neurotransmitter’s based on the

behavior of acetylcholine, but in recent years, there have been several, what they call,
unconventional neurotransmitters identified. They're considered neuromodulatory.

They aren't packaged in vesicles or released by exocytosis. They're synthesized and

released during synaptic transmission instead of beforehand. And they're oftentimes
in response to another neurotransmitter release. They are modulators and they may
even be providing feedback.

One of the things that's interesting about them is that they're hydrophobic, so they're
able to go right through the cell membrane, and interact with proteins inside both
the presynaptic and the postsynaptic neurons.

Examples of these are gases like nitric oxide and carbon monoxide, and lipid
metabolites like endocannabinoids, which I'll talk a little bit more about near the end
of the episode. So, what does cause a neurotransmitter to be released?

In a conventional neurotransmitter, the vesicles merge with the cell membrane and
release the neurotransmitter into the synapse, which is called exocytosis, and this
requires an influx of calcium. But in the end, everything comes down to the
neurotransmitter receptors. At least a hundred genes have been identified that code
for different receptors.

The types of receptors, again, are under the same as the membrane types of receptors
that are present in all cells; ionotropic, ligand-gated, or neurotransmitter-gated ion
channels that have a fast effect. And then metabotropic GE-protein-coupled ones
that are slow, indirect, and neuromodulatory, and most neurotransmitters interact
with both types of receptors.

[Music]

So, I do want to talk just briefly about the structure and function of the two types of
receptors. The ionotropic receptors are pretty simple. They have a place called the
extracellular binding domain, which is where the neurotransmitter binds and then
the trans-membrane ion channel.

6 © Copyright 2023 Virginia A Campbell, MD

 Now, some ionotropic receptors are more complex, and the NMDA receptor is an
example of this. However, the metabotropic G-protein-coupled receptors are much
more complex because they use the second messenger and there's a wide variety of
effects that include indirectly opening or closing ion channels, but can go as far as
turning genes on and off.

One situation where ionotropic receptors actually use a second messenger is they
open a calcium channel, and then that calcium comes in and acts as a second
messenger. And when this happens, calcium can actually act as a second messenger
by regulating a wide variety of proteins.

The influx can cause small excitatory signals that are called excitatory postsynaptic
potentials (EPSP) as opposed to when the negative ion chloride comes in, it causes
what's called inhibitory postsynaptic potential (IPSP).

I must mention that communication between neurons is not limited to synaptic

transmission. There's another kind of transmission called volume transmission, such
as when neuromodulators are released into the extracellular space or the CSF and
the targets are outside the synapse.

This was first described in the 1980s because they had noticed that there was a
mismatch between where they found the receptors and where neurotransmitters
were being released, and the receptors were being found far away from the known
release places.

Now, volume transmission can involve a conventional neurotransmitter or an

unconventional neurotransmitter, or even a neuropeptide. Some examples of things
that might have volume transmission; neurohormones, neurotropic factors, and
immune modulators.

Some specific examples: there’s something called glutamate spillover, and the
endocannabinoids actually do their neuromodulation via volume transmission. In
fact, they seem to bind to presynaptic receptors.

But the key idea is that neurotransmitters and neuropeptides can have effects that
happen far away from where they were actually released. And I mentioned that
calcium influx pushes the cell toward depolarization while chloride ions push it in
the opposite direction.

So, the excitatory and inhibitory postsynaptic potentials are summing up both
temporally and spatially. And so, when a threshold is reached, then an action
potential happens and is transmitted, and also, synaptic transmission not fixed. And
we'll talk a little bit more about how plasticity happens when we talk with Seth Grant
next month.

So, it turns out that the receptors are more interesting and complex than the
neurotransmitters. Most of the so-called conventional neurotransmitters are
relatively small molecules that occur even in single cell life forms. Outside the
context of the nervous system, they have a wide variety of activities.

7 © Copyright 2023 Virginia A Campbell, MD

 I'm afraid that if I did a detailed description of the main neurotransmitters and
neuropeptides, your eyes would start to glaze over. So, I'm going to try to provide a
high-level view and use acetylcholine as examples.

Remember that the two types of receptors are ionotropic and metabotropic, I
mentioned that it's the receptors that determine what happens. It's also interesting to
know that the target neurons may have both types of receptors, and this makes sense
if you remember that the ionotropic receptors act quickly and the metabotropic
receptors have slower, more varied modulatory actions reactions.

Let's talk about acetylcholine in a little bit more detail. And as I mentioned, you want
to go to the book, The War of the Soups and the Sparks to learn more about how this
was all figured out.

If a neuron releases acetylcholine, it's called a cholinergic neuron, and you'll often
hear doctors talk about anticholinergic side effects, and that is what it's referring to,
is things that affect the receptors for acetylcholine.

There are two types of cholinergic receptors, one called nicotinic and one called
muscarinic. These names reflect the fact that they were discovered before there was a
distinction appreciated between the ionotropic and metabotropic receptors.

The nicotinic receptors are ionotropic, they're activated by nicotine, which is a plant
alkaloid. There's two types, muscle type and neuronal type. And the neuronal type is
the one that is 50 times more sensitive to nicotine. They're all non-selective cation
channels permeable to both sodium and potassium.

Some antagonists are dextromethorphan, tubocurarine, and toxins from snake

venom. Muscarinic receptors are the metabotropic G-protein-coupled receptors.
There's five subtypes. These actually regulate both ion channels and second
messenger pathways involving kinases and phosphatases which are enzymes.

So, there's a wide variety of downstream effects. Some are short-term and some are
long-term. They affect everything from membrane potentials to gene expression. The
agonists, they're activated by muscarine, which is a mushroom toxin. In the
peripheral nervous system, the targets are of all postganglionic parasympathetic
neurons and a few sympathetic neurons have these muscarinic receptors.

The CNS has a widespread concentration of receptors. The preganglionic neurons in

the spinal cord release acetylcholine at the adrenal medulla, which then releases
epinephrine as a hormone, and then the lower motor neurons in the spinal cord
release acetylcholine at the neuromuscular junction.

The neuromuscular junction is the junction between the lower motor neuron from
the spinal cord and the skeletal muscle, and was the first and best characterized
chemical synapse. It's excitatory. When the action potential in the presynaptic
neuron reaches the axon terminal, it activates the voltage-gated calcium channels
and allows calcium to flow in down its concentration gradient into the cell.

It's the calcium that causes the vesicles that contain the acetylcholine to merge with
the membrane and release acetylcholine into the synapse. The nicotinic receptor

8 © Copyright 2023 Virginia A Campbell, MD

 opens sodium channels, which causes a depolarization to about minus 40 millivolts.

This travels passively along the muscle, allowing additional voltage-gated sodium
channels to open, which eventually, triggers the action potential.

The action potential allows more calcium to flow in, and this is what triggers the
steps that lead to muscle contraction, which is why if you don't have any calcium, you
have no muscle contraction.

It's also important to know that acetylcholine is broken down by an enzyme called
acetylcholinesterase, which is what allows the contraction to end. Reversible anti-
cholinesterase drugs are actually being used to treat early Alzheimer's disease, but
the esterase is also targeted by insecticides and so-called nerve gases like sarin gas.

This is why organophosphate poisoning is so dangerous. Because acetylcholine has a

role throughout the nervous system, it's not surprising that it's been implicated in a
wide variety of disorders, including the psychotic symptoms of schizophrenia and
Parkinson's disease, and the cognitive deficits of dementia.

Now, I'm not really going to be focusing on these clinical aspects for several reasons.
One is that the details get overwhelming pretty quickly, but the main reason is that
our understanding of the actual mechanisms involved is very poor and often,
speculative.

For example, the idea that depression or any other mental illness is due to a shortage
or excess of one or more neurotransmitters has been disproven long ago, but still
hangs around. In fact, even as we've progressed from this primitive idea, why the
various drugs work is still very poorly understood.

So, I prefer to focus on what we do know, and I want to emphasize again, it's the
receptor that actually determines what happens. One major problem with designing
drugs is that even if they're aimed at the receptors, they almost always have
unintended side effects.

Even a particular type of metabotropic receptor triggers different pathways and some
of those might be best left alone. But before I leave acetylcholine, I want to mention
two more important roles.

So, the cholinergic acetylcholine is important in the autonomic nervous system. All
the presynaptic neurons are cholinergic, all the postsynaptic parasympathetic
neurons are cholinergic. The presynaptic neurons interact with rapid acting nicotinic
receptors. The postsynaptic parasympathetic neurons interact with muscarinic
receptors with a wide variety of effects.

Additionally, in recent years, the enteric nervous system has been classified as a
separate division of the peripheral nervous system. These neurons function
autonomously to control the GI tract. The enteric nervous system also uses
cholinergic neurons.

In addition to our new appreciation for the enteric nervous system, we now also
appreciate that cholinergic neurons are important in the brain. The three main areas
are the basal forebrain, basal ganglia, and brainstem.

9 © Copyright 2023 Virginia A Campbell, MD

 But from there, projections go everywhere. It appears that a loss of cholinergic

neurons projecting to the neocortex and hippocampus are one of the earliest changes
of Alzheimer's disease. The spinal cord involves both types of receptors, and
cholinergic transmission in the CNS is mostly neuromodulatory.

So, this might be a time we could consider the question: why is nicotine so
addictive? This has to do with the role of nicotinic receptors in the reward circuits of
the brain. The normal role is to encourage people or animals to repeat rewarding
behavior like eating.

But nicotine also creates a false signal that the activity such as smoking is rewarding
even when it's not. And this is why most people consider intentionally putting
nicotine in products unethical. It's been proven that the tobacco industry suppressed
the evidence that nicotine was addictive for many, many years.

So, I think we now have a little bit of time left to talk about neurotransmitters whose
names might be familiar. Feel free to take a break if you want to, and come back.

One key thing to remember is that all the so-called conventional neurotransmitters
are small molecules that are either simple amino acids or derived from a single
amino acid. Two important examples that are simple amino acids are glutamate and
GABA.

Glutamate is the main excitatory neurotransmitter, while GABA is the main

inhibitory neurotransmitter. Glycine also appears to be an inhibitory transmitter in
the spinal cord, but participates in excitatory functions in the CNS such as the
coactivation of the NMDA receptor.

So, GABA has both ionotropic and metabotropic or G-protein-coupled receptors. The
ionotropic ones are ligand-gated chloride channels. Remember, chloride is negative,
so when it enters the cell, it pushes it away from depolarization.

That's why GABA is an inhibitory neurotransmitter, so-called GABA agonists. That is

drugs that interact with the GABA receptors include barbiturates, ethanol,
benzodiazepines, and drugs like Ambien. Drugs like AmbienTM can actually be
hallucinatory, and I have personally experienced that effect.

Now, there are also the GABA-B receptors, which are metabotropic G-protein-
coupled receptors. They seem to be involved in the oscillatory activity of the brain,
and their dysregulation is associated with substance abuse, anxiety, and depression.

Neurons that release glutamate are called glutaminergic, and they're usually
excitatory. They're the most abundant in the brain, so they affect most aspects of
brain function. Glutamate is the main excitatory neurotransmitter in the mammalian
brain.

It has both ionotropic and metabotropic G-protein-coupled receptors. I think it's

actually kind of interesting that the NMDA receptor’s actually ionotropic, even
though it's really very complex, it's not just a simple ion channel.

[Music]

10 © Copyright 2023 Virginia A Campbell, MD

 Many familiar neurotransmitters are classified as monoamines, and this just means
that they contain a single amine group. So, the so-called catecholamines are derived
from the amino acid, tyrosine. The pathway is tyrosine to L-DOPA to dopamine, to
norepinephrine, to epinephrine.

Dopamine is the one that everybody has kind of heard of as the reward
neurotransmitter, which is very inaccurate. The dopamine neurons are located in the
midbrain region of the brain stem in the substantia nigra and the ventral tegmental
area and in the hypothalamus, but they get then projected all over the brain.

The substantia nigra is involved in voluntary movement and it's particularly

vulnerable to loss of neurons in Parkinson's disease. Dopamine is involved in at least
two reward pathways, but it's important to realize that dopamine is involved in
much, much more. Like every other neurotransmitter, its effect depends on which
receptor it activates as well as the location of that receptor.

Dopamine has important roles outside the nervous system, and it's released by non-
neuronal cells. It reduces GI motility and protects GI mucosa. It inhibits insulin
synthesis in the pancreas. It increases sodium secretion, and urine output in the
kidneys.

Dopamine has five subtypes of G-protein-coupled receptors that are divided into D1-
like and D2-like. The D1 type are the mostly postsynaptic. They can be excitatory or
inhibitory, and they're the most abundant receptor in the human.

The D2-like is also located either pre or postsynaptically, usually inhibitory. And it’s
the dysregulation of the D2 type that's associated with schizophrenia and bipolar
disorders.

Then it has to also be removed like any other neurotransmitter. And there's disorders
that involve deficits in the transport or removal of dopamine. And this can also be
targeted by drugs of abuse and by therapeutics.

So, we go from dopamine, which then can be synthesized to norepinephrine. The

norepinephrine neurons are located in the brainstem in three particular brainstem
regions, and then they project all over the brain (see a theme here?) They're involved
in sleep and dreaming, attention, emotion, cognition.

They have similar triggers to the things that trigger the sympathetic nervous system.
That is to mobilize the body for action. In the peripherals nervous system, remember
that the preganglionic neurons are going to release acetylcholine, but most of the
postganglionic sympathetic neurons are going to then release norepinephrine to the
various organ systems leading to constriction of the pupils, regulation of heart rate
and blood flow, and also going to the adrenal medulla, which will then release
norepinephrine and epinephrine.

And remember, that once these are released, since they're released into the
bloodstream, at that point, the norepinephrine and epinephrine are not acting as
neurotransmitters. They're acting as hormones. In the body, they cause
vasoconstriction and increased heart rate.

11 © Copyright 2023 Virginia A Campbell, MD

 From the norepinephrine, you can synthesize the epinephrine, which was originally
called adrenaline. Its effects are pretty similar to norepinephrine, but also, include
sexual arousal, appetite, and metabolic control. And then again, remember that the
sympathetic activity would then lead to both release of both norepinephrine and
epinephrine.

The receptors for norepinephrine and epinephrine were actually discovered before
the appreciation of the classification system that I described before. But they're
basically metabotropic G-protein-coupled receptors, and they come in alpha and beta
types. They're expressed in many regions of the brain. The beta-1 dominates in the
cortex and the beta-2 receptors in the cerebellum. They have a variety of
neuromodulatory effects.

I have to mention that when I was in medical school, we learned about beta receptors
in the context of blood vessels and airways, and neurotransmitters weren't even on
our radar. In the periphery, these determine the effects of sympathetic nervous
systems. The alpha receptors mediate smooth muscle contraction, the beta receptors
are mediating heart muscle contraction, smooth muscle relaxation, and
gluconeogenesis.

So, drugs that target these systems have a wide variety of applications from heart-
related conditions to mental health problems. And it's not hard to imagine that they
are also affected by drugs of abuse as well as causing unintended side effects.

An example of a drug would be a beta blocker might be used for blood pressure. Now,
even beta blockers are used for treatment, but because of all this stuff with the
receptors, this is the reason why modern pharmacology is trying to make their drugs
more targeted by trying to find the subtypes, and then make their drugs more
selective by targeting the subtypes.

So, the older drugs tend to be what they call a non-selective. So, propranolol, for
example, which was the first beta blocker, is very non-selective. It's going to block
any beta receptor.

Now, still on the monoamines, we've got serotonin and histamine. Serotonin is
another one that you've probably heard about since there's drugs that have serotonin
in their name because they're serotonin reuptake inhibitors.

Serotonin is synthesized from the amino acid, tryptophan and it's also known as 5
hydroxytryptamine (5-HT) So, you often see it called 5-HT in instead of serotonin.
So, it's kind of a trick to remember that they're the same thing.

Serotonergic neurons are located in nine nuclei in the reticular formation in the
brainstem, so lower than some of the other ones we've talked about. And this is not
surprising when you consider that it's one of the oldest neurotransmitters from an
evolutionary standpoint, and it gets projected, these serotonergic neurons from the
reticular activating system go all over the brain.

It's involved in a wide variety of functions including mood regulation, pain

processing, and homeostatic mechanisms like appetite, thermoregulation, energy
balance, and just basically, the hypothalamic pituitary adrenal access.

12 © Copyright 2023 Virginia A Campbell, MD

 There are seven families of receptors for serotonin, and only one of these is
ionotropic. The rest are metabotropic G-protein-coupled receptors, and they mediate
their effects, their actions through the release of other neurotransmitters.

The one that's an ionotropic one is called 5-HT3. It's a non-selective cation channel
that's permeable to sodium, potassium, and calcium. I mention it because it's
important. These receptors are actually localized to the part of the brainstem that
controls nausea and vomiting, and drugs that block this 5-HT3R receptor like
ZofranTM are very important for treating and preventing nausea.

These receptors are also located in other parts of the brain and their mutations are
associated with bipolar disease, depression, anorexia, and irritable bowel. Some of
the other serotonin receptors are associated with functions like memory and blood
pressure and control.

One of the receptors, the 5-HT2R was first noted as a target for psychedelic drugs
like LSD. So, you may have heard that serotonin receptors play a role in depression,
since the particular class of drugs is called the serotonin reuptake inhibitors, which
means it targets the uptake.

But I again, want to emphasize that how these drugs work is not really understood
and the serotonins receptors also seem to be targeted by drugs of abuse. I want to
emphasize several things.

First, it's possibly because of its ancient origins serotonin has a role in a wide variety
of body functions related to homeostasis. Interestingly, the highest concentrations of
5-HT are actually produced in the GI tract.

It also has a role in the cardiovascular system where it may cause vasoconstriction or
vasodilation depending on the receptor type. This is a very strong demonstration of
the principle I've been emphasizing throughout this episode.

In the end, everything comes down to the receptors. Life seems to have been using a
handful of small molecules since it emerged millions of years ago, and long before
nervous systems appeared. We have explored this with Seth Grant in the past and
we'll explore it further with him next month.

Before I talk briefly about neuropeptides and unconventional neurotransmitters, I

want to mention histamine, because it's another monoamine neurotransmitter that
you probably associate with allergic reactions since it has an important role in the
immune system, and it's also released in the GI tract.

Histaminergic neurons are located in the posterior hypothalamus, and these go to

several regions in the cortex. There are four types of metabotropic G-protein-coupled
receptors for histamine.

H1 receptors increase wakefulness and prevent sleep. The histamine neurons actually
stop firing completely during sleep, which is kind of interesting. And so, when you
get a drowsiness from taking an antihistamine like BenadrylTM, it's because of its
effects on the HA1 receptor. And the newer antihistamines that don't make you

13 © Copyright 2023 Virginia A Campbell, MD

 sleepy are ones that target different histamine receptors and miss this one that does
this.

Before we finish, I want to spend a few minutes talking about neuropeptides and
unconventional neurotransmitters. I mentioned both of these earlier on, but I want
to just go back over them briefly. The neuropeptides, what is a neuropeptide? A
neuropeptide is just a small peptide, 3 to about 40 amino acids in length that acts
like a neurotransmitter, really more like a neuromodulator.

So, what we have is in the human genome, we have genes that code for about 90
precursors that can be processed into about a hundred neuropeptides. So, actually,
when you hear that there's hundred neurotransmitters, it's not really true. Really, the
basic conventional neurotransmitters was a pretty small batch.

But then we have all these other substances that are neuropeptides. So, most neurons
are going to make at least one conventional neurotransmitter, say glutamate or
GABA. And then one or more neuropeptide, which is going to be a co-transmitter,
which modulates the effect of the basic neurotransmitter.

Neuropeptides are similar to peptide hormones. Well, actually, they're the same
exact thing except it's about where they're released. Same compound, if it's released
in the synapse or extracellular space, be called a neuropeptide. If it's in the
bloodstream, it gets called a hormone.

And an example of this is oxytocin, and I'll give you some more examples in a
minute. So, how is a neuropeptide different from a neurotransmitter? First of all, it's
not synthesized at the synapse, it's synthesized in the cell body in a place called the
rough endoplasmic reticulum.

And then usually, it gets released away from what's called the active zone of the
synapse. And very importantly, it does not get recycled back into the presynaptic
neuron. And this means that it can wander off and do other things away from where
it's released, which is very important, and it can have a more prolonged action. Many
of the neuropeptides were originally discovered in the context of regulation of
hormone release, and this includes many pituitary hormones.

Neuropeptides activate metabotropic G-coupled-protein receptors, and they usually

act as neuromodulators. In fact, they are mostly acting as co-transmitters with
conventional small neurotransmitters, but their effects can include modulation of
gene expression.

Neuropeptides are distributed throughout the brain, but they've been most
extensively studied in the hypothalamus, which is an ancient highly conserved brain
structure involved in homeostasis. When we say it's conserved, it means that the
human hypothalamus is very similar to that of other mammals and even other
vertebrates.

So, let's think of some examples of neuropeptides. The most abundant is one you
probably haven't heard of. It's called neuropeptide Y. It is made up of 36 amino
acids, functions in both the central nervous system, and the peripheral nervous
system.

14 © Copyright 2023 Virginia A Campbell, MD

 In the peripheral nervous system, it's produced by sympathetic neurons along with
norepinephrine causing vasoconstriction among other effects. Its highest levels in the
hypothalamus and five different receptors have been identified that will interact with
neuropeptide.

Why? It has many roles including promoted food intake and fat storage.

Now, you've probably heard that the brain makes its own opioids. These are also
neuropeptides, and they appear to be made by special set of neurons in the
hypothalamus, and then get distributed throughout the brain. They're often released
with other conventional neurotransmitters.

Some additional examples of neuropeptides are the hypothalamic hormones like

oxytocin, and vasopressin, substance P, and glucagon. So, you may recognize some of
these things as being hormones.

So, just remember that you can't tell from the name what a polypeptide does since
the same molecule will be considered a hormone if it's released into the bloodstream,
but act as a neuromodulator if it's released within the nervous system.

This is one reason why we are learning that the immune system and the nervous
system are highly integrated. If you listen to last month's episode about the
entangled brain, you may get a sense of an ongoing theme.

Scientists break the body down into separate systems, but most of these systems
interact, and this interaction leads to a level of complexity that is not always
appreciated either by scientists or laypeople.

[Music]

So, what about unconventional neurotransmitters? This means the so-called

gasotransmitters and endocannabinoids. They are not synthesized, stored or released
by presynaptic neurons. That's why they're unconventional. They're actually
hydrophobic and they're usually synthesized in response to a conventional
neurotransmitter. They actually diffuse directly across the membrane into the target
that's inside the cell.

Okay, so an unconventional neurotransmitter's not going to have a receptor on the

cell membrane like you normally think of. They can go inside, they go into the
presynaptic, the postsynaptic, or the adjacent neurons.

Gases like nitric oxide seem to be made in a regulated manner and have specific
functions at the physiological level. Nitric oxide’s actually been implicated in
plasticity, and abnormal nitrous oxide signaling may have a role in
neurodegenerative diseases.

Now, at least two endogenous endocannabinoids have been identified; anandamide

and 2-arachidonoylglycerol, which is usually called 2-AG. These are lipids that are
produced by the postsynaptic neuron in response to neurotransmitters that increase
calcium influx. So, they get released into the extracellular space and then they bind to
the cannabinoid receptors.

15 © Copyright 2023 Virginia A Campbell, MD

 There are two types of metabotropic G-protein-coupled receptors that have been
identified. The CB1 in the neuro CNS and the CB2 in the peripheral nervous system.
And THC, which comes from cannabis is also binding to these receptors, which many
of you recognize the cannabinoid part of the name.

Studies have shown that the CB1 receptors at the presynaptic membrane inhibit
neurotransmitter release. The CB2 receptors also have functions in the immune
system. Endocannabinoids and THC affect regulation of appetite, eating, sleep, pain,
relief, motivation, and pleasure. Anandamide and THC can impair memory and adult
neogenesis in rodent models.

Now, obviously, there's a need for more research about both an endogenous and
exogenous cannabinoids.

One interesting thing to note is that although the endocannabinoids obviously have a
physiologic function, they are not conventional neurotransmitters. There's no such
thing as a cannabinoid neuron. Instead, they're released by along with a conventional
neurotransmitter. And since they interact with the metabotropic receptors, their
effects are somewhat indirect.

On the other hand, as lipids, they can diffuse directly across the cell membrane, and
this may account for their rapid action when used recreationally. And it may also
explain why the topical agents seem to be effective. Because in order to get a drug to
go transdermally across your skin, it needs to be a lipid or tied to a lipid in some way.

Now, I realize that many of you are interested in learning more about cannabinoids,
and this has not been a very deep discussion. It may not even be the most up-to-date
since I used a textbook for it. Unfortunately, as a part of my move, some of my other
books on this subject are not accessible to me at this time. But I will look for the
opportunity to explore this topic further when I get a well-referenced book on the
topic.

Before I review some of the key ideas, I just have a few brief announcements. The
first one is a special request for those of you who enjoy YouTube, please subscribe to
the Brain Science Podcast YouTube channel, and then actually listen to at least one
full episode. This will help me meet the requirements for monetization. I won't make
very much, but every little bit helps.

I will try to keep my announcements short today, but I do want to remind you that
today's show notes include an extensive list of books and previous episodes for those
who want to learn more about topics like Neuroanatomy and Action Potentials.

Although it's not listed in this month's show notes, I also want to mention that Luiz
Pessoa’s new book, The Entangled Brain is a great book for both new listeners and
even for those of you who were around the last time I talked about neurotransmitters
back in 2007. You'll find these show notes in your audio app or at
brainsciencepodcast.com, and please do send me feedback at
brainsciencepodcast@gmail.com.

16 © Copyright 2023 Virginia A Campbell, MD

 You can also get the show notes for free in the Brain Science mobile app, which is
now called Brain Science Podcast. This app is available for all mobile devices and is a
great way to access both free and premium content.

Now, for an update on my move to New Zealand — I was expecting to be there by the
end of May 2023, but it took over six months for the medical counsel of New Zealand
to approve my palliative medicine credentials. That finally happened in June, so I
was finally able to apply for my work Visa, and I now, hope to be in Auckland by mid-
August 2023.

Unfortunately, that means I had to give away my tickets for the Women's World Cup.
It has been fantastic hearing from listeners from both Australia and New Zealand.
Please do reach out to me if you haven't already done so. My email is
brainsciencepodcast@gmail.com.
Wherever you live, you can be sure that you never miss an episode of Brain Science
by signing up for the free Brain Science newsletter. That way, you get show notes
automatically every month. Just text BrainScience (all one word) to 55444.
BrainScience (all one word) to 55444.

Also, since Brain Science is independently produced, it relies on listeners like you for
financial support. Your support literally keeps Brain Science on the air. I want to
thank everyone who supports my work, either financially or by sharing it with others.

I know that some of you find it confusing to choose between the various MyLibsyn
Premium and Patreon, so I've created a table at brainsciencepodcast.com/premium
that compares these so you can pick the one that's best for you. And of course,
donations are always okay too.

I have every intention of continuing to produce Brain Science after I move to New
Zealand, so please support the show in whatever way you can, and don't forget to
subscribe and listen on YouTube if you can. Obviously, this was a fairly limited
discussion of the fascinating topic of neurotransmitters.

I started out by discussing some of the basics of brain anatomy and neuron structure,
along with a very brief discussion of how an action potential is produced. If you want
to learn more about these topics, I highly recommend going to
brainsciencepodcast.com and looking at the show notes or looking at the show notes
in your audio app, presuming that the app preserves the links. The show notes
include both books and links to previous episodes on these topics.

The meat of this episode was a discussion on neurotransmitters and their receptors.
Most neurotransmitters are small molecules that have functions outside the neuro
system. If a molecule, histamine, for example, is released into the blood stream, it
functions as a hormone, but it's only considered neurotransmitter if it's released by a
neuron and interacts with a specific receptor usually on a different neuron.

The key to the incredible flexibility and diversity of neurotransmitters comes down to
the increasing complexity and diversification of their receptors, and we will explore
this from an evolutionary perspective next month with Seth Grant.

17 © Copyright 2023 Virginia A Campbell, MD

 Despite their diversity, the receptors come in two basic types; ionotropic or ligand-
gated, and metabotropic G-protein-coupled. It is easy to remember which is which by
remembering that the ionotropic receptors are actually ion channels that are opened
and closed when the ligand, that is to say the neurotransmitter interacts with a
receptor protein in the cell membrane.

Since this is a direct effect, it can occur fairly quickly on the order of a 10th of a
millisecond. In contrast, the metabotropic G-protein-coupled receptors use one or
more second messengers to kick off everything from indirectly opening and closing
channels to changing gene expression.

And while the actual synaptic transmission is only slowed down to about 0.5 to 3
milliseconds, the actual effects may occur much later, especially in the case of gene
regulation.

Thus, this interaction with these receptors is called neuromodulation. Obviously, the
most basic categorization of neuron activity is to ask whether it is excitatory or
inhibitory. This depends on which ion channels are opened or closed. Anything that
allows a positive cations such as sodium to flow into the cell will make it less negative
until a threshold is reached, and then the action potential fires.

In contrast, if negative anions like chloride are allowed to enter the cell, it becomes
more negative, and it's inhibited from triggering an action potential. Obviously, any
individual neuron is receiving anywhere from thousand to 10,000 individual signals,
and these sum up over time and space, and determine whether the neuron
depolarizes.

Glutamate is the most common excitatory neurotransmitter. It has several ionotropic

receptors that are non-selective cation channels that produce fast excitatory
responses.

In contrast, GABA binds to an ionotropic receptor that allows the ion chloride to
come in, which inhibits the polarization by making the cell more negative.

This might give you the impression that there is a one-to-one correspondence
between each neurotransmitter and what it does, but this is not true because
metabotropic G-protein-coupled receptors are much more diverse than that.

And like many neurotransmitters, glutamate interacts with both types of receptors.
The monoamines like histamine, dopamine, norepinephrine, and epinephrine, only
have metabotropic G-protein-coupled receptors. And from my review, it appears that
glycine is the only one that has only ionotropic receptors.

Depending on your background, you may have found this material either too basic or
completely overwhelming. Whatever your background, I want you to focus on a few
key ideas.

The most important one is that each neurotransmitter has many complex functions
that are determined by which receptors that it binds to, and where these receptors
are located. It is inaccurate to describe any neurotransmission by a single action.

18 © Copyright 2023 Virginia A Campbell, MD

 For example, do not think of dopamine as the reward molecule, because although it
is important in the reward circuits of the brain, that's not the only thing that it does,
and that really is probably the same message that I communicated back way back in
episode eight.

It's all about the receptors. I mean, it's actually kind of surprising how simple the
actual neurotransmitters are, and as we will talk further with Seth Grant, it's
probably because these simple molecules have been having a signaling role since the
beginning of life and before the evolution of nervous systems.

I hope that this episode has given you a sense of how this works. Feel free to let me
know what errors I made as I'm sure I made some. I try to be accurate, but this is a
complex subject, and I won't be offended if those of you who know more about it
than I do point out if I made a mistake.

But I do think that the key idea that the average person needs to know is that the
receptors are where the action is, and if you hear somebody tell you that dopamine
does this or serotonin does that, you know that that is an oversimplification, and
might even be totally wrong.

Next month, I'm going to be talking with Seth Grant some more about the synapse,
its evolution, and his more recent discoveries about how diverse synapses really are
in terms of which molecules like the receptors, which ones are actually there is very,
very varied. So, you'll want to be sure to come back for that next month.

In the meantime, I hope you'll check out my other podcasts, Books and Ideas and
Graying Rainbows., and send me email at brainsciencepodcast@gmail.com.

Thanks again for listening. I look forward to talking with you again next month.

[Music]
Brain Science is copyrighted to Virginia Campbell, MD. You may copy this episode to
share it with others, but for any other uses or derivatives, please contact me at
brainsciencepodcast@gmail.com.

The theme music for Brain Science is “Mind Fire,” written and performed by Tony
Cotraccia. You can find his work at syncopationnow.com.

19 © Copyright 2023 Virginia A Campbell, MD