Brain Neurotransmitters

I. INTRODUCTORY REMARKS

Most of the previous chapters of this series have emphasized gross organization and
structure of the brain. This has been essential in order to gain perspective, but from
a cryonicist's point of view preservation of the "the anatomical basis of mind" will
ultimately mean preservation of the structures only visible under a microscope.
Understanding what structures to look-for and how those structures might best be
preserved is the ultimate goal of this series.

As a step in the direction towards understanding finer structure, this chapter will
examine the brain from a more chemical point of view than the previous installments
— with particular reference to the gross anatomy and function of neurotransmitters in
the brain.

II. THE CHEMICAL ENVIRONMENT OF THE BRAIN

Skull surface dissection

The brain has the consistency of firm jelly, and therefore is protectively encased in a
thick, bony skull. The brain literally floats in about 150 millilitres (mL)
of CerebroSpinal Fluid (CSF) secreted by the choroid plexus. Approximately 500
mL of CSF is secreted daily, which slowly circulates down through the four
ventricles, up through the subarachnoid space and exits into the cerebral veins through
the arachnoid villi. The brain has no lymphatic system, so the CSF serves as a partial
substitute. 

Skull section

The dura mater is a tough, protective connective tissue which is tightly bound to the
skull, but which encases the cerebral veins. Under the dura mater is the subarachnoid
space containing CSF, arteries and web-like strands of connective/supportive tissue
called the arachnoid ("spider- Brain ventricles
like") mater. The pia mater is a
permeable membrane of collagen, elastin fibers & fibroblasts on the floor of the
subarachnoid space which allows diffusion between the CSF and the interstitial fluid
of the brain tissue. The pia mater lies on a membrane that is infiltrated with astrocyte
processes. The dura mater, the arachnoid mater and the pia mater are collectively
referred-to as the meninges. 

Brain ventricles
 Skull Medial Cross-Section showing CSF flow

While the brain & CSF

are separated by the
somewhat permeable pia
mater, the blood-
cerebrospinal fluid
barrier and the blood-
brain barrier (BBB)
represent substantial
protection for the brain
against undesirable
blood substances. These
barriers are very
permeable to water,
oxygen, carbon dioxide
and small lipid-soluble
substances. They are
also somewhat
permeable to small
electrolytes — and
special transport systems
exist for some other
specific molecules such as essential amino acids. The barriers are the result of
endothelial cells which line capillary walls — and glial cells called astrocytes which
wrap the capillaries with fibers.

The brain is not only a functionally distinct organ, it is a chemically distinct one. 50%
of dry brain weight is lipid (in contrast to 6-20% for other organs). Most of the brain
lipid is structural (in myelin or membranes) in contrast to the triglycerides and free
fatty acids constituting the fat of other organs. The blood-brain barrier creates a
protected chemical environment for the brain wherein certain molecules can perform
functions independent of the functions those molecules perform in the rest of the
body. This is particularly important for the neurotransmitters serotonin (which is
highly concentrated in platelets & the intestine) and norepinephrine (which affects
blood pressure & metabolism). All of the known amino-acid neurotransmitters
are non-essentialamino acids. This means that they can be manufactured in the brain,
without needing to be supplied from outside the brain. But in the major area of the
brain which does not have a blood-brain barrier — the hypothalamus — the primary
neurotransmitters are peptides. 

III. GENERAL COMMENTS ABOUT NEUROTRANSMITTERS

The three major categories of substances that act as neurotransmitters are (1) amino
acids (primarily glutamic acid, GABA, aspartic acid & glycine),
(2) peptides (vasopressin, somatostatin, neurotensin, etc.) and
(3) monoamines (norepinephrine, dopamine & serotonin) plus acetylcholine. The
major "workhorse" neurotransmitters of the brain are glutamic acid (=glutamate) and
GABA. The monoamines & acetylcholine perform specialized modulating functions,
often confined to specific structures. The peptides perform specialized functions in the
hypothalamus or act as co-factors elsewhere in the brain. [For a well-organized
categorization of neurotransmitters, see Neurotransmitter (Wikipedia).]

Although there are many neurotransmitters in the central nervous system, the
peripheral nervous system has only two: acetylcholine and norepinephrine. Why are
there so many brain neurotransmitters? Because the functions performed by brain
neurotransmitters are not as uniform as they might superficially appear. Some (like
glutamate) are excitatory, whereas others (like GABA) are primarily inhibitory. In
many cases (as with dopamine) it is the receptor which determines whether the
transmitter is excitatory or inhibitory. Receptors can also determine whether a
transmitter acts rapidly by direct action on an ion channel (eg, nicotinic acetylcholine
receptors) or slowly, by a second-messenger system that allows for synaptic plasticity
(eg, muscarinic acetylcholine receptors). Speed & mechanism of transmitter
inactivation after the signal has been sent is also a factor. There are probably also
costs & benefits involved in synthesizing, transporting and recycling various
neurotranmitters in the differing chemical
mileus of the brain.

Many of these issues will become more

clear in discussing the synthesis, distribution
and function of the major brain
neurotransmitters.

IV. GLYCINE
 Glycine

Glycine is the simplest of amino acids, consisting

of an amino group and a carboxyl (acidic) group
attached to a carbon atom. Glycine's function as a
neurotransmitter is also fairly simple. When
released into a synapse, glycine binds to a
receptor which makes the post-synaptic
membrane more permeable to Cl- ion. This
hyperpolarizes the membrane, making it less
likely to depolarize. Thus, glycine is an inhibitory
neurotransmitter. It is de-activated in the synapse
by a simple process of reabsorption by active
transport back into the pre-synaptic membrane.

Glycine is a neurotransmitter only in vertebrate animals. The glycine receptor is

primarily found in the ventral spinal cord. Strychnine is a glycine antagonist which
can bind to the glycine receptor without opening the chloride ion-channel (ie, it
inhibits inhibition). The resultant spinal hyperexcitability is what makes strychnine a
poison. Quoting from the ENCYCLOPEDIA BRITANNICA:

"Symptoms of poisoning usually appear within 20 minutes, starting with stiffness at

the back of the neck, twitching of the muscles, and a feeling of impending suffocation.
The patient is then seized with violent tetanic convulsions in which the body is arched
and the head bent backward. After a minute the muscles relax, and the patient sinks
back exhausted, heightened perceptiveness being perceived throughout due to sensory
cortex stimulation. A touch, a noise or some other stimulus causes the convulsions to
recur; or they may recur spontaneously, often at intervals of a few minutes. Strychnine
poisoning is ultimately the result of suffocation or exhaustion."

V. ASPARTIC ACID (ASPARTATE)

Like glycine, apartate is primarily localized to the Aspartate

ventral spinal cord. Like glycine, aspartate opens
an ion-channel and is inactivated by reabsorption
into the pre-synaptic membrane. Unlike glycine,
however, apartate is
an excitatory neurotransmitter, which increases
the likelihood of depolarization in the
postsynaptic membrane. Aspartate & glycine
form an excitatory/inhibitory pair in the ventral
spinal cord comparable to the
excitatory/inhibitory pair formed by glutamate & GABA in the brain. Interestingly,
the two exitatory amino acids — glutamic acid & aspartic acid — are the
two acidic amino acids found in proteins, insofar as both have two carboxyl groups
rather than one.
VI. GLUTAMIC ACID (GLUTAMATE)

Glutamate is the most common

neurotransmitter in the brain. It is always Glutamate
excitatory, usually due to simple receptors
that increase the flow of positive ions by
opening ion-channels. Glutamate
stimulation is terminated by a (chloride-
independent) membrane transport system
that is only used for re-absorbing
glutamate & aspartate across the pre-
synaptic membrane. Glutamate &
aspartate re-enter the cell by a transporter
driven by the high extracellular concentrations of Na + and the high intracellular
concentrations of K+. Soduim enters the cell along with the amino acids and potassium
leaves the cell — much the way a pulley couples the lifting of a light weight with the
fall of a heavier weight. Thus, glutamate/asparate entry is indirectly powered by the
ATP-driven Na+-K+-ase (sodium pump) which creates the high ion concentration
gradients.

Possibly the most complicated of all neurotransmitter receptors is the NMDA

glutamate receptor. N-Methyl-D-Aspartate is a synthetic chemical not naturally
found in biological systems, but it binds specifically to the NMDA glutamate receptor
(receptors are frequently named for artificial substances that bind to the receptor with
higher specificity than their natural neurotransmitter ligands). The NMDA receptor is
the only known receptor which is regulated both by a ligand (glutamate) and by
voltage. There are at least 5 binding sites which regulate NMDA receptor activity, ie,
sites for (1) glutamate (2) glycine (3) magnesium (4) zinc and (5) a site that binds the
hallucinogenic substance phencyclidine (PCP, "angel dust"). Phencyclidine can
induce psychosis — an NMDA effect that is difficult to explain. NMDA receptors
have a capacity for an activity-dependent increase in synaptic efficiency known as
LTP (Long-Term Potentiation), which may be crucial to some forms of learning &
memory. Inhibition of NMDA activity (and LTP) is believed to be an important part
of the wayethanol affects brain functions.

NMDA receptors are most densely concentrated in the cerebral cortex (hippocampus,
especially — particularly the CA1 region), amygdala, & basal ganglia. They are
particularly vulnerable to glutamic acid excitotoxicity, ie, damaging effects due to
excessive excitatory neurotransmitter release. Both aspartic acid & glutamic acid (the
two amino acids having 2 carboxyl groups — the "acidic amino acids") have the
capacity for destroying neurons when released in excessive amounts (although
calcium seems to be more of a cause than acidity). Monosodium glutamate (MSG),
a major component of soya sauce, has been shown to destroy nerve cells when fed to
young animals. Insofar as glutamate does not normally cross the blood-brain barrier, it
is open to question whether this is relevant to a human adult. Increased alertness (or
anxiety) due to caffeine may be mainly due to blockage of adenosine receptors which
normally inhibit glutamate release.

Glutamate released into synapses is either reabsorbed directly into neurons by the ion-
exchange transport system described above, or is soaked-up by astrocytes (glial cells)
which convert the glutamate into glutamine (a molecule which cannot cause
excitotoxicity). The glutamine can then be safely transported back to neurons for re-
conversion into glutamate. One of the damaging effects of mercury poisoning is
swelling of astrocytes, which are rendered unable to soak-up glutamine from synapses
(contributing to excitotoxicity). Excitotoxicity due to glutamic acid is a major
destructive process seen in stokes and other forms of brain ischemia (see Ischemia and
Reperfusion Injury in Cryonics).

Nitric oxide can act as neuromodulator when glutamate stimulation of NMDA

receptors results in nitric oxide synthesis & release — enhancing neurotransmitter
release from adjacent synapses. Granule cells of the dentate gyrus of the
hippocampus are rich in nitric oxide synthetase. Nitric oxide may contribute to LTP.

VII. GAMMA AMINO BUTYRIC ACID (GABA)

GABA Biochemistry

GABA is the major inhibitory neurotransmitter of the brain, occurring in 30-40% of

all synapses (second only to glutamate as a major brain neurotransmitter). It is most
highly concentrated in the substantia nigra & globus pallidus nuclei of the basal
ganglia, followed by the hypothalamus, the periaqueductal grey matter ("central
grey") and the hippocampus. The GABA concentration in the brain is 200-1000 times
greater than that of the monoamines or acetylcholine.

GABA is somewhat unique among neurotransmitters insofar as it is commonly

inactivated (after release into the synapse) by active transport into the astrocyte glial
cells that are closely associated with synapses. Both glutamate and GABA are
synthesized in the brain from the Krebs citric acid molecule alpha-keto glutarate — a
reaction known as the "GABA shunt". GABA is synthesized from glutamic acid and
is catabolized back into the citric acid cycle. The vitamin B6 derivative pyridoxal
phosphate is a cofactor in the synthesis of GABA, which is why seizures occur in
Vitamin B6 deficiency. GABA levels rise when the citric acid cycle activity is low
(ie, when cell energy usage is low), and the resultant generalized GABA inhibitory
effect on the brain neurons can be protective during hypoxia or ischemia.
Like glycine, the GABA receptor is connected to a chloride ion channel, allowing
more chloride ion to enter the cell and thus making the membrane less likely to
depolarize. A closely associated receptor site will bind to benzodiazepines (such as
diazepam) to increase the frequency of channel opening. Caffeine can neutralize the
effects of benzodiazepine tranquilizers such as diazepam
(Valium®). Benzodiazepines act by enhancing the effect of GABA on
GABAA receptors, whereas caffeine has an opposite effect by inhibiting GABA
release. Barbiturates slightly decrease the frequency of opening, but prolong the
duration. The benzodiazepine receptor site is thought to be the natural site of action of
a yet-unidentified peptide. By potentiating the effects of GABA, the benzodiazepines
function as so-called "minor tranquilizers" (to be distinguished from the anti-
psychotic "major tranquilizers"). Anxiety is the most frequently diagnosed psychiatric
disorder — affecting 10-30% of people — which is why diazepam (Valium) was for
many years the most frequently prescribed drug in North America. Alcohol &
barbiturates have similar effects on the GABA receptor. In fact, potentiation of
chloride influx into neurons is a major mechanism in the effect of ethanol on the
brain. Some of the effects of benzodiazepines are probably due to GABA synapses on
monoamine-producing neurons. GABA receptors can also be blocked, and the
insecticide dieldrin is used for this purpose.

Prolonged use of benzodiazepines results in adaptation of the receptors to their use.

Receptors may increase in number and/or sensitivity to GABA. (An increase or
decrease in receptor number or sensitivity due to receptor alteration by drugs is known
as upregulation or downregulation, respectively. A larger dose of benzodiazepine
may be needed to produce the same result — a phenomenon known as tolerance.
Withdrawal of the drug can result in GABA receptor hypoactivity producing
symptoms worse than the ones that the patient originally sought treatment for. Such
symptoms are called withdrawal. The phenomenon of receptor adaptation and
drug dependence is seen with most drugs that act at synapses, including ones that are
excitatory or potentiating as well as inhibitory or deactivating.

VIII. ACETYLCHOLINE

Acetylcholine was the first Acetylcholine

neurotransmitter discovered and is the
major neurotransmitter in the peripheral
nervous system (the only other peripheral
neurotransmitter being norepinephrine).
Acetylcholine is usually (but not always)
an exitatory neurotransmitter — in
contrast to the monoamine
neurotransmitters, which are nearly always
(with a few exceptions) inhibitory. Acetylcholine in the brain is produced
from acetyl-CoA, resulting from glucose metabolism, and from choline, which is
actively transported across the blood-brain barrier. Most dietary choline comes
from phosphatidyl choline, the major phospholipid in the membranes of
plants&animals (but not bacteria). The acetyl-CoA & choline are independently
synthesized in the neuron cell body and independently transported along the axon to
the synapse where they are conjugated into acetylcholine.

There are comparatively few acetylcholine receptors in the brain, but outside the brain
acetylcholine is the major neurotransmitter controlling the muscles. Body muscles can
be divided into the skeletal muscles system (under voluntary control) and the smooth
muscles of the autonomic nervous system (controlling heart, stomach, etc.
— not under voluntary control). The autonomic nervous system is further subdivided
into sympathetic and parasympathetic divisions. Direct innervation of skeletal
muscles is due to acetylcholine, as is the innervation of smooth muscles of the
parasympathetic nervous system. Direct innervation of the sympathetic nervous
system (except for sweat glands) is due to norepinephrine (or both epinephrine &
norepinephrine in the case of the adrenal medulla). 

Sympathetic and Parasympathetic Nervous Systems

The sympathetic nervous system innervates body organs in "fight or flight" situations,
so the role of norepinephrine as the end-organ neurotransmitter should not be
surprising. End-organ stimulation by acetylcholine in the parasympathetic nervous
system is more "vegetative", eg, assisting digestion. Acetylcholine receptors are of
two types: (1) a fast-acting ion-channel controlled receptor and (2) a slow-acting
receptor that acts through a G-protein (Guanine nucleotide-binding protein) that
stimulates second-messengers (often cyclic AMP) to indirectly open ion-channels.
Direct ion-channel controlling receptors can respond in microseconds, whereas
indirect second-messenger controlling receptors take milliseconds to produce a
response. Only indirect, second-messenger controlling receptors have the capacity for
plasticity. The two acetylcholine receptor classes are named for artificial toxins that
selectively activate them. The fast-acting receptor is named nicotinic, because it is
specifically activated by the toxin found in tobacco. The slow-acting receptor is
named muscarinic, because the toxin muscarine (found in poisonous mushrooms) and
acetylcholine will activate it, but nicotine will not.

Parasympathetic nerves are either cranial or sacral. 75% of all parasympathetic fibers
arise from a single cranial nerve: the vagus nerve. These fibers travel to end-organs
containing ganglia. It is the short postganglionic nerves from the ganglia to the
smooth muscles in the end-organs which are muscarinic. The preganglionic fibers are
nicotinic. Similarly, the preganglionic fibers of the sympathetic nervous system are
nicotinic, although the sympathetic ganglia exist as distinct nodules closer to the
spinal cord. The neuromuscular junction of skeletal muscles is also nicotinic.

Considering the rapidity with which skeletal muscles must often be able to respond to
the volition to move, it is understandable that they are controlled by fast-acting
nicotinic receptors. Unlike other neurotransmitters (which rely on re-uptake),
acetylcholine activity in both muscarinic and nicotinic synapses is primarily stopped
by an enzyme, ie, acetylcholinesterase. For nicotinic synapses, this means that a signal
can be both rapidly initiated and rapidly terminated. The choline resulting from the
hydrolysis of acetylcholine can be transported across the presynaptic membrane for
resynthesis into acetylcholine.

Some snake venoms contain toxins that block nicotinic receptors, thereby paralyzing
their victims. Similarly, some South American Indians used the nicotinic blocking
agent curare (extracted from plants) as a poison on their arrowheads. Atropine,
which blocks muscarinic receptors, is also a poison. But atropine-like substances are
of use for dilating the eye through topical application, for examination of the retina. 

Not only are there

relatively few
cholinergic neurons in
the brain, but their
distribution is "spotty"
— in contrast to the
monoamines which have
distinct midbrain nuclei
serving as the major
sources of brain
innervation. Most brain
cholinergic receptors are
muscarinic, which may
make sense insofar as
only second-messenger
controlled receptors are
capable of synaptic
plasticity. The site of
greatest acetylcholine
synthesis in the brain is
the interpeduncular nucleus (located near the substantia nigra in the midbrain). All
of the interneurons in the striatum (caudate nucleus & putamen of the basal ganglia)
and the nucleus accumbensare cholinergic. The septum provides cholinergic fibers
to the septal-hippocampal tract. The primary cholinergic input to the cerebral cortex
comes from the basal nucleus of Meynert, which is the most prominent structure in
the substantia innominata(ventral to the anterior half of the globus pallidus, and
adjacent to the hypothalamus). Meynert's nucleus also innervates the basolateral
amygdala, the basal ganglia and the reticular nucleus of the thalamus. Cholinergic
input from the basal nucleus to the cerebral cortex is active in both the waking state
and in REM sleep, but is reduced in non-REM sleep.

Administration of centrally-acting muscarinic blocking agents results in memory loss

for normal individuals. The fact thatAlzheimer's Disease patients show severe
reduction in Meynert's nucleus neurons has led to the suspicion that cholinergic loss
could be an important factor in memory loss for these patients. Benefits from the
anticholinesterase drugs have reinforced this belief. But Alzheimer's patients have
neurofibrillary tangles, loss of NMDA receptors and reduced numbers of
noradrenergic & serotonergic neurons as well. Some Alzheimer's patients show
normal cholinergic cell numbers in Meynert's nucleus.

Centrally-acting anticholinesterases such as physostigmine (which can cross the

blood-brain barrier — in contrast to neostigmine, which cannot) will worsen the
tremors seen in Parkinson's Disease. Conversely, atropine-like substances (which are
anti-muscarinic) reduce those tremors. These effects are clearly a consequence of the
cholinergic interneurons in the striatum. Physostigmine & neostigmine are known
as reversible anticholinesterases because they bind to acetylcholinesterase, but will
eventually disengage. This makes them of therapeutic use for glaucoma & myasthenia
gravis. Irreversible anticholinesteraseswhich permanently bind-to and inactivate
acetylcholinesterase, are only of use as insecticides. The only treatment for poisoning
by these substances is atropinic drugs.

Stimulation of brain nicotinic receptors by nicotine from tobacco and arecoline from

betal nuts (chewed by a quarter of the population of India) is a source of euphoric
addiction for a large segment of the world's population. Research funded by the
tobacco industry has shown that working memory may be improved by nicotinic
stimulation of dopaminergic neurons in the substantia nigra & ventral tegmental area
[BRAIN RESEARCH 657:165-170 (1994)] and that nicotine may be able to improve
learning & memory in Alzheimer's patients showing a loss of nicotinic receptors in
the neocortex & hippocampus [PHARMACOLOGY BIOCHEMISTRY AND
BEHAVIOR 52:517-523 (1995)].

IX. DOPAMINE

The primary monoamine neurotransmitters are dopamine, norepinephrine and

serotonin. Dopamine and norepinephrine are catecholeamines, whereas serotonin is an
indolamine.
The amino acid tyrosine is not an essential amino acid because it can be synthesized
in the liver from phenylalanine by the enzyme phenylalanine hydroxylase. But it
cannot be synthesized in the brain, and therefore must enter the brain by the large
neutral amino acid transporter, which also transports phenylalanine, tryptophan,
methionine and the branch-chained amino acids. These amino acids all compete for
the transporter, so a large quantity of one of the other amino acids in the blood stream
could greatly limit the amount of tyrosine entering the brain. One case in which this
occurs is when there is a liver deficiency of phenylalanine hydroxylase. In that case,
Phenylalanine reaches high concentrations in the blood and monopolizes the large
neutral amino acid transporter, producing the mental retardation of phenylketonuria. 

Dopamine synthesis

Once in the brain, tyrosine can be converted to DihydrOxyPhenylAlanine (DOPA) by

the tyrosine hydroxylase enzyme using oxygen, iron and TetraHydroBiopterin (THB)
as co-factors. High concentrations of dopamine inhibit tyrosine hydroxylase activity
through an influence on the THB co-factor. DOPA is converted to dopamine
by Aromatic Amino Acid Decarboxylase (which is fairly nonspecific insofar as it will
decarboxylate any aromatic amino acid) using PyridoxaL Phosphate (PLP) as a co-
factor. This reaction is virtually instantaneous unless there is a Vitamin B6 deficiency.

Dopamine & epinephrine are primarily inhibitory neurotransmitters that produce

arousal. This may sound paradoxical, but the most likely explanation for this effect is
that the postsynaptic cells for catecholamines themselves are inhibitory. There are 3-4
times more dopaminergic cells in the CNS than adrenergic cells. Dopamine in the
caudate nucleus facilitates posture, whereas dopamine in the nucleus accumbens is
associated with an
animal's speed (and
pleasure). 

There are two primary

dopamine receptor-
types: D1 (stimulatory)
and D2 (inhibitory), both
of which act through G-
proteins. D2receptors
often occur on the
dopaminergic neurons,
partially for the purpose
of providing negative
feedback. These so-called autoreceptors can inhibit both dopamine synthesis and
release.

The binding of dopamine to D1-receptors stimulates the activity of Adenylyl

Cyclase (AC), which converts ATP to cyclic AMP (cAMP), a second
messenger which binds to Protein Kinase A (PKA). PKA then modulates the activity
of various proteins by the addition of phosphate. 

There are 4 main

dopaminergic tracts in
the brain:
(1) the nigrostriatial
tract from the substantia
nigra to the striatum
accounts for most of the
brain's dopamine
(2) the tuberoinfundibu
lar tract from the
arcuate nucleus of the
hypothalamus to the
pituitary stalk, which
has a controlling effect
on the release of the
hormones prolactin thro
ugh tonic inhibition via
D2 receptors
(3) the mesolimbic
tract from the ventral
tegmental area to many
parts of the limbic
system and
(4) the mesocortical
tract from the ventral
tegmental area to the
neocortex, particularly
the prefrontal area.
Dopamine cells project
topographically to the
areas they innervate. 

Both dopamine &

norepinephrine are
catabolized by a two-
step process involving
the
enzymes MonoAmineOxidase (MAO) and Catechol-O-MethylTransferase (COMT).
COMT is primarily active in the synapses, and uses &S-Adenosyl Methionine (SAM)
as a methyl-group donor. MAO is primarily active in the pre-synaptic terminal against
catecholamines that are not safely enclosed in storage vesicles. Normally, COMT only
catabolizes about 10% of synaptic catecholamine, since catecholamine synaptic
activity is primarily terminated by re-uptake into the pre-synaptic neuron terminal.
MAO accounts for a much larger portion of catecholamine metabolism.

The darkly pigmented neurons in the pars compacta of the substantia nigra accounts
for 80% of the dopamine in the brain. The dark pigment neuromelanin is a dopamine
polymer that makes the substantia nigra appear black. Motor control in the striatum
(caudate nucleus and putamen) is thought to involve a balance between inhibitory
dopaminergic (D2) and excitatory cholinergic neurons.

A form of MAO, known as MAO-B, is the most common form of MAO in the
striatum. MAO-B is known to metabolize the neurotoxin MPTP to its active form.
When striatum dopamine is depleted to 20% the original level, symptoms of
Parkinson's Disease appear. Administration of DOPA is the most common treatment.
The monoamine oxidase inhibitor deprenyl will specifically inhibit MAO-B, thereby
making deprenyl a useful adjunct to DOPA therapy. The ergot
derivative bromocriptine is a D2 agonist which can alleviate the symptoms.
(Deprenyl's reputed anti-aging capabilities may be related to MAO-B inhibition.
Similarly, carnosine, which has been shown to reduce cellular senescence, inhibits
MAO-B.) 

Phenothiazine derivatives

Schizophrenia is thought to be due to an overstimulation of D 2 receptors in the

mesolimbic and mesocortical systems. Evidence for the "excess dopamine" theory of
schizophrenia comes largely from the fact that D 2 antagonist drugs alleviate the
symptoms, whereas substances which increase D2 stimulation, such as amphetamines,
can induce psychotic symptoms (which are reversible with D 2antagonists). About 10%
of Parkinsonian patients given DOPA treatment will develop psychotic symptoms
resembling schizophrenia.

The major classes of antipsychotic drugs are the phenothiazines (eg, chlorpromazine),
the butyrophenones (eg, haloperidol) and the thioxanthenes (eg, chlorprothixene).
Butyrophenones are 100 times more potent against D 2 receptors than against
D1 receptors. The similarity in shape between a portion of the chlorpromazine
molecule and dopamine indicates how chlorpromazine could bind to a dopamine
receptor without triggering a response.

The mesolimbic & mesocortical dopaminergic systems are thought to play an

important role in motivation, by attaching cognition of incentive significance to
stimuli. In experiments on animals that are motivated to electrically self-stimulate
themselves with electrodes implanted in their brains, dopamine is the mediating
neurotransmitter for the locus ceruleus, lateral hypothalamus, ventral tegmental area
and sulcal prefrontal cortex (but not the nucleus accumbens or substantia nigra). Fruit
flies that are "socially stimulated" have three times the amount of dopamine in their
brains than "socially deprived" fruit flies — an effect that correlates directly with
more sleep (presumably consolidating learning) in the stimulated flies [SCIENCE;
Indrani,G; 313:1775-1781 (2006)].

Cocaine particularly increases dopaminergic activity in the mesolimbic areas of the

brain by inhibiting dopamine re-uptake in the ventral tegmental area and the nucleus
accumbens. Amphetamine seems more generalized in its action, not only by inhibiting
re-uptake, but by releasing dopamine from most brain regions. Both cocaine &
amphetamine produce feelings of psychological energy & arousal, associated with
diminished appetite & need for sleep. Both cocaine & amphetamine can lead to visual
& tactile hallucinations as well as paranoid thinking, although the psychotic effects of
amphetamine may also be mediated by increased serotonin release. Chronic
amphetamine users seem to lose a capacity for normal pleasure — which has been
correlated with neuron degeneration in the mesolimbic area.

Perception of time-intervals is believed to be mediated by spiny neurons located in

the striatum of the basal ganglia. Timing begins with a burst of dopamine and ends
with a recognized signal. Marijuana slows subjective time by lowering dopamine
available, whereas cocaine and methamphetamine accelerates the sense of time by
increasing dopamine availability. (Adrenaline and stress hormones can also "make
seconds feel like hours".)

The natural brain amine phenylethylamine (PEA, found in chocolate) has been
associated with the "love-excitement" of sexual attraction & emotional infatuation.

PEA concentrations are normally highest in the nucleus accumbens (a "reward

center") followed by the frontal & cingulate cortices. Levels spike during orgasm and
ovulation. PEA is very similar to amphetamine in chemical structure and may
likewise act by causing dopamine release, but endorphin release may be a significant
effect. PEA is preferentially oxidized by MonoAmine Oxidase-B (MAO-B), which
may account for the anti-depressant effects of selegiline.

Other actions of dopamine include the induction of vomiting by stimulation of

D2 cells in the chemoreceptor trigger zone, stimulation of growth hormone release by
D2 receptors, and increased exploration & locomotion (thought to be connected to
dopaminergic activity in the nucleus accumbens). Sexual behavior in the male is
increased by dopamine agonists, whereas sexual behavior in the female is increased
by dopamine antagonists.

X. NOREPINEPHRINE (NORADRENALIN)
 Noradrenalin Synthesis from Dopamine

Norepinephrine (along with acetylcholine) is one of the two neurotransmitters in the

peripheral nervous system. Norepinephrine is synthesized from dopamine by means of
the enzyme Dopamine Beta-Hydroxylase (DBH), with oxygen, copper and Vitamin C
as co-factors. Dopamine is synthesized in the cytoplasm, but norepinephrine is
synthesized in the neurotransmitter storage vesicles. Cells that use norepinephrine for
formation of epinephrine use SAMe (S-AdenylMethionine) as a methyl group donor.
Levels of epinephrine in the CNS are only about 10% of the levels of norepinephrine.

The most prominent noradrenergic (ie, norepinephrine-containing) nucleus is

the locus ceruleus in the pons, which account for over 40% of noradrenergic neurons
in the rat brain. Most of the other noradrenergic neurons are clustered in a region
described as the lateral tegmental area. The neocortex, hippocampus, and
cerebellum receive noradrenergic stimulation exclusively from the locus ceruleus.
Most of the dopaminergic innervation of the hypothalamus comes from the lateral
tegmental nuclei. 

Electrical stimulation of
the locus ceruleus
produces a state of
heightened arousal. The
noradrenergic system is
most active in the awake
state, and it seems to be
important for focused
attention, in contrast to
the motor arousal of
dopamine. Although the
locus ceruleus has been
identified as a pleasure
center, it also seems to
contribute to anxiety.
Increased neuronal
activity of the locus
ceruleus is seen upon
the occurrence of unexpected sensory events. Brain norepinephrine turnover is
increased in conditions of stress. Benzodiazepines, the primary antianxiety drugs,
decrease firing in the locus ceruleus, thus reducing distribution of noradrenalin to the
forebrain and amygdala. This is part of the explanation for the use of benzodiazepines
for inducing sleep.

Active projection of norepinephrine from the locus coeruleus of the reticular

activating system to the forebrain is a key feature of awakeness-arousal as
distinguished from sleep. Norepineprhine projection to the basal nucleus of the
forebrain is low in sleep — virtually absent in REM (Rapid Eye-Movement) sleep.
The basal nucleus when stimulated by norepinephrine from the locus coeruleus sends
neuromodulating acetylcholine to the cerebral cortex, thereby promoting alertness.

The beta-adrenergic blocking drug propranolol has also been used to treat anxiety. By
blocking the adrenergic inputs to the amygdala, beta-blockers inhibit the formation of
traumatic memories. Cortisol stimulation of the locus coeruleus due to chronic stress
exacerbates norepinephrine stimulation of the amygdala. Healthy humans fed fish oil
rich in the omega-3 fatty acids EPA and DHA show reduced plasma levels of the
stress-associated hormone norepinephrine [NUTRITION; Hamazaki,K; 21(6):705-
710 (2005)].

Beta-noradrenergic receptors also apparently inhibit feeding, whereas alpha-receptors

seem to stimulate feeding.

Although MAO inhibitors reduce metabolism of all catecholamines, it is believed that

the anti-depressant effect is more related to norepinephrine than to dopamine. Most
MAO in the brain is of type-B, but drugs selective for inhibiting MAO-A have proven
to be better anti-depressants. MAO-A preferentially metabolizes norepinephrine &
serotonin. MAO-A inhibiting drugs given for depression have critically elevated
blood pressure in patients eating tyramine-containing foods (such as cheese) due to
the failure to metabolize tyramine (which can act as a pressor agent). These drugs (eg,
phenelzine & pargyline) inactivate MAO by forming irreversible covalent bonds.
More modern MAO inhibitors are safer because they form reversible bonds. MAO-B
inhibitors like deprenyl are also less likely to cause the "cheese effect". (Alcohol also
selectively inhibits MAO-B.) 
Tricyclic Antidepressants
Tricyclic anti-depressants derive their name from their 3-ring structure. Desipramine
only inhibits norepinephrine re-uptake, with little effect on dopamine. Imipramine &
amitriptyline are inhibitors of norepinephrine and serotonin re-uptake by the
presynaptic terminals, but are more potent for serotonin. Cocaine is also a potent
inhibitor of catecholamine re-uptake, but it does not act as an anti-depressant. Weight
gain due to increased appetite is a frequent side effect of tricyclic anti-depressants,
particularly of amitrip- tyline. By contrast, both cocaine & amphetamine reduce
appetite.

Both MAO inhibitors and tricyclic anti-depressants have immediate effects on brain
monoamines, but clinically anti-depressants require several weeks of administration
before they produce a therapeutic effect. It is therefore believed that it is not the
immediate effects on neurotransmitters that is producing the antidepression, but the
long-term effects on modification of receptors.

Excessive cortisol secretion is seen in 40-60% of depressed patients, associated with

diminished noradrenergic inhibition of corticotropin-releasing hormone secretion in
the hypothalamus. Corticotropin-releasing hormone induces anxiety in experimental
animals.

XI. SEROTONIN (5-HYDROXYTRYPTAMINE, 5-HT)

Serotonin was isolated from the blood serum as a

substance causing powerful smooth muscle
contraction. Only later was it demonstrated to be
tryptamine with a hydroxyl group at the 5-position.
Only 1-2% of the serotonin in the body is in the brain,
insofar as serotonin is widely distributed in platelets,
mast cells, etc. But there is no equilibration between
body serotonin and brain serotonin — the serotonin in
the brain is independently synthesized from tryptophan
transported across the blood-brain barrier. 

Serotonin synthesis

Serotonin synthesis is a 2-step process, the first step of which requires the
enzyme tryptophan hydroxylase with oxygen, iron and THB as co-factors. Neither
the enzyme nor the co-factors are rate-limiting for either step of these reactions —
virtually all brain tryptophan is converted to serotonin. Serotonin concentration in the
brain is far more sensitive to the effects of diet than any other monoamine
neurotransmitter — and can be increased up to 10-fold by dietary supplementation in
laboratory animals.

Consumption of a meal that is high in carbohydrate, branch-chained amino acids and

tryptophan has a particularly dramatic effect because both glucose from carbohydrate
and branch-chained amino acids (especially leucine) increase insulin secretion. Insulin
facilitates the transport of the branch-chained amino acids into muscle cells, thereby
reducing the competition tryptophan faces for the large neutral amino acid transporter
that takes it across the blood-brain barrier. The resultant drowsiness induced by
serotonin is a common effect of a large carbohydrate meal. 

The richest concentration of serotonin in the body can be found in the pineal body,
even though this gland does not use serotonin as a neurotransmitter. Instead, serotonin
is primarily methylated in the synthesis
of melatonin. Melatonin derives its name Melatonin
from the fact that it can darken the skin of
amphibians ("melas" is Greek for "black") — although it has also been reported to
induce pigment lightening in cells. Melatonin is synthesized from serotonin in a 2-step
process that takes an acetyl group from acetyl-CoA and a methyl group from
by SAMe (S−Adenosyl Methionine).

Melatonin is of particular importance for regulating diurnal (circadian) & seasonal

behavior & physiology in mammals. The pineal body has been called a "third eye"
because its activity is influenced by light. In mammals, noradrenergic neurons near
the optic nerve are inhibited by light. In darkness, norepinephrine stimulation of
pineal cells causes the release of cyclic AMP second-messenger, which activates
(phosphorylates) the N-acetyl transferase enzyme which catalyzes acetylation of
serotonin. In many specied (including humans) melatonin is an inhibitor of sexual
activity in both sexes.
Decreased melatonin in
the Spring leads to
rutting — and the birth
of offspring in the
warmer seasons.
Melatonin also
stimulates production
of brown adipose
tissue, a special form of
fat which (when burned)
only produces heat, not
ATP. This is especially
important for
hibernating animals. 
Serotonin neurotransmitter neurons are located in the raphe nuclei. The caudal
(closer to the "tail") nucleus projects largely to the medulla and spinal cord for the
regulation of pain perception. The rostral (closer to the "beak") nucleus projects
extensively to the limbic structures and the cerebral cortex. In the limbic system,
especially, the projections are co-localized with norepinephrine receptors — and the
two transmitters seem to work in conjunction in the regulation of arousal.

The SupraChiasmatic Nucleus (SCN) of the hypothalamus regulates the mammalian

circadian clock ("day-night cycles"), partially in response to light. Melatonin release
is inhibited as a result of the response of the SCN to light. The SCN is richly
innervated by serotonergic input from the dorsal raphe nucleus. Serotonin inhibits the
responsiveness of the SCN (and thus the circadian rhythm) to light [ANNALS OF
MEDICINE 31:12-33 (1999)]. Sleep deprivation increases serotonin release in the
SCN [BRAIN RESEARCH 909:81-91 (2001)]. With aging there is a decline in both
serotonin transporters [LIFE SCIENCES; Yamamoto,M; 71(7):751-757 (2002)] and
serotonin receptors [NEUROPSYCHOPHARMACOLOGY; Meltzer,MD; 71(7):751-
757 (2002)]. Depletion of serotonin is believed to be related to the disruption of the
circadian rhythm associated with senescence [AMERICAN JOURNAL OF
PHYSIOLOGY 272(2 Pt 2):R509-R513 (1997)].

As little as one or two grams of L−tryptophan is effective in decreasing sleep latency

time [PSYCHOPHARMACOLOGY; Schneider-Helmert,D; 89(1):1-7 (1986) and
PHARMACOPSYCHIATRY; Demisch,K; 20(6):242-244 (1987)]. L−tryptophan
either improves or has not effect on other sleep parameters, with the exception of a
suppressive effect on REM sleep [EUROPEAN NEUROLOGY; Korner,E;
25(Suppl 2):75-81 (1986)].

Almost all antidepressant treatments (tricyclic antidepressants, monoamine oxidase

inhibitors, lithium, electoconvulsive therapy, serotonin reuptake inhibitors, etc.)
augment serotonin neurotransmission [MOLECULAR PSYCHIATRY; Bauer,M;
7(2):140-156 (2002)]. Low levels of serotonin are associated with high levels of pain
sensitivity, locomotion, aggression, and sexual activity [BIOLOGICAL
PSYCHIATRY; Lucki,I; 44(3):151-162 (1998)]. Low levels of serotonin are also
associated with depression, panic disorders, and Obsessive-Compulsive
Disorder (OCD). Patients with evidence of low serotonin levels have attempted
suicide by very dramatic means, such as cutting the throat. This may explain some of
the therapeutic effects offluoxetine (Prozac), which selectively prevents the re-uptake
of serotonin, thereby increasing synaptic serotonin concentrations (elevated
serotonin). Fluoxetine is also distinctive because it has a half-life of about four days.
Fluoxetine has been used therapeutically for panic, obsessive-compulsive and eating
disorders (such as bulimia). Unlike the tricyclic anti-depressants, which often
stimulate appetite, fluoxetine more often reduces appetite. Fluoxetine may even
enhance learning [PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR
52:341-346 (1995)]. Depression patients treated with tryptophan as well as fluoxetine
show less sleep disturbance at the outset of treatment than patients treated with
fluoxetine alone [JOURNAL OF PSYCHIATRY & NEUROSCIENCE; Levitan,RD;
25(4):337-346 (2000)].
Rats subjected to learned helplessness conditioning (electric shocks) became
desensitized to serotonin, but exercise (running) reduces this effect [JOURNAL OF
NEUROSCIENCE; Greenwood,BN; 23(7):2889-2898 (2003)]. Depression of
hippocampal neurogenesis by learned helplessness conditioning in rats can be
reversed by fluoxetine [NEUROPSYCHOPHARMACOLOGY; Malberg,JE;
28(9):1562-1571 (2003)].

The natural carbohydrate inositol has been used to elevate serotonin without some of

the side effects of fluvoxamine (Luvox, which is similar to Prozac). A dose of
18 grams/day inositol has been shown to be effective against Obsessive-Compulsive
Disorder (OCD) [THE AMERICAN JOURNAL OF PSYCHIATRY; Fux,M;
153(9):1219-1221 (1996)]. The same dose of myo-inositol has been shown to be as
effective as flavoxamine against panic disorders [JOURNAL OF CLINICAL
PSYCHOPHARMACOLOGY; Palatnik,A; 21(3):335-339 (2001)]. Cantaloupe,
oranges, and grapefruit are the foods containing the highest inositol concentrations,
but to get 18 grams/day inositol would require eating 13 cantaloupes, 45 grapefruits,
or 58 oranges [AMERICAN JOURNAL OF CLINICAL NUTRITION; Clements,RS;
33(9):1954-1967 (1980)].

The tricyclic antidepressants typically inhibit both norepinephrine & serotonin re-
uptake by pre-synaptic terminals. The effectiveness of these neurotransmitters against
depression seems to be due to both decreased functional activity of beta-postsynaptic
norepinephrine receptors and increased activity of type−2 serotonin receptors in the
limbic regions of the brain. But experimental depletion of monoamines only leads to
depression in subjects having a family history of this disorder [MOLECULAR
PSYCHIATRY; Ruhe,HG; 12(4):331-359 (2007)].

Monkeys with high levels of testosterone & low levels of serotonin are both
aggressive & lacking in restraints on impulsive/violent behavior. Arsonists who
commit their crime for mercenary reasons show normal levels of serotonin, but those
who commit the crime impulsively have low serotonin. Lead interferes with serotonin
synapse formation. Monkeys experimentally exposed to lead became so dangerously
aggressive that the study was halted early [CHEMICAL & ENGINEERING NEWS
81(22):33-37 (2003)].

Reserpine prevents the transport of all the monoamines (and acetylcholine) into
storage vesicles in the presynaptic membrane — leaving them vulnerable to
destruction by monoamine oxidase. Reserpine (as extracts from the Rauwolfia plant)
was used for centuries in India to treat "hysteria". Reserpine has been used as a potent
tranquilizer, but it can produce serious depression that may lead to suicide attempts.

LySergic acid Diethylamine (LSD) acts most strongly on the type-2 serotonin

receptors, but it also has some effect on norepinephrine receptors. Serotonin seems to
play a role in dreaming. During both dreaming and LSD intoxication, electrical
activity in the visual cortex arises from the brain stem rather than from the eyes. LSD
not only induces visual hallucinations, but it heightens sensory awareness while
diminishing control of sensory input. The reduced ability to distinguish between
sensory impressions can lead to feelings of being "in union with the universe".
Artificial stimulation of the raphe simulates the actions of LSD, decreasing
habituation to repetitive stimuli. Low doses of LSD & amphetamine, however, have
been shown to enhance a form of associative learning.

High-estrogen contraceptives may have contributed to depression by lowering

serotonin levels in the brain. Low levels of growth hormone in depressed patients may
be due either to low levels of norepinephrine, serotonin, or both.

XII. PEPTIDES

Peptides are the most common neurotransmitters in the hypothalamus. Their complex
structure can allow for high receptor specificity. They are all synthesized on
ribosomes and are all inactivated by hydrolysis at the synapse (rather than by re-
uptake). Peptides are far more potent than other neurotransmitters, requiring only very
small amounts to produce a profound effect. Even very minute amounts of
somatostatin can inhibit growth hormone release.

Opioid peptides include the endorphins, enkephalins and dynorphins. Enkephalins

are frequently found in presynaptic (axo-axonic) synapses. Opiates and enkephalins
(or endorphins) inhibit the firing of locus ceruleus neurons. The highest concentration
of opioid receptors are found in the sensory, limbic and hypothalamic regions of the
brain — and are particularly high in the amygdala & periaqueductal grey area.
Opioids tend to be released as slower-acting co-transmitters which modulate the
action of the associated neurotransmitter (such as glutamate) which is being released
from the same synapse. Although opioids are generally inhibitory, they have an
excitatory effect on hippocampal pyrimidal neurons mediated by inhibition of GABA
release.

Cholecystokinin (CCK) seems to function in the production of satiety. Injection of

small quantities of this peptide into the ventricles or the paraventricular nucleus can
inhibit feeding. CCK is associated with dopamine synapses in some limbic areas, and
appears to modulate dopamine release. Such peptide synergy with other transmitters is
common. For example, GABA is often associated with somatostatin and serotonin
with Substance P.

Low doses of the peptide vasopressin have been shown to enhance learning in
laboratory animals. However, humans with vasopressin deficiency show no signs of
memory impairment. Because vasopressin is potent in increasing blood pressure, its
use by humans should be approached with caution. Safer analogues may yet be found.

XIII. RELEVANCE TO CRYONICS

The profound effects of neurotransmitters on the human psyche seems to portend

substantial implications for personal identity. But their obvious emphemeral nature
makes it seem unlikely that they can be described as being part of "the anatomical
basis of mind". Water is also an important component of the human brain, but water is
very uniform and replaceable. Preservation of the means of producing, storing,
releasing and re-uptaking neurotransmitters (ie, the receptors and synaptic
connections) seems more important than preserving the neurotransmitters themselves.
The fact that anti-depressants & anti-psychotics take weeks to become effective may
offer some clue in this regard.

In a sense, however, many of the "structural" features of neurons are no less

ephemeral than neurotransmitters. They can be produced or replaced by enzymes
coded-for by DNA in much the same way as neurotransmitters. This would seem to
point to neuron gene expression as the key locus for identity, and the most crucial site
for preservation. But gene expression is determined by the mileu of the neuron — a
circular homeostasis that seemingly does not allow for finding "ultimate causes".
Moreover, considering the demonstrably critical role played by astrocytes in GABA
re-uptake at synapses, one might wonder whether glial cells are also an essential
component of identity.

(For more on the subject of the locus of consciousness in the brain and the relevance
to preservation of consciousness see my essay Neurophysiology and Mental
Function.)