Chapter 4

Chemical Signaling in the Nervous System

T.M. Dwyer

coordinate specific neurotransmitters in conveying information

The Synapse: Adaptable to Many Tasks-54
Structure and Function-54
Transmitters and Modulators-54
Synthesis and Storage-54
Release-55
Transduction-55
Termination-55
Glutamate Receptors: The Tetrameric Excitatory
Ionotropic Receptors-55
AMPA Receptors and Milliseconds of Excitation-56
NMDA Receptors and Seconds of Activation-59
Kainate Receptors: AMPA-like, but Presynaptic-59
Cys-Loop Receptors: The Pentameric
Multifunctional Ionotropic Receptors-59
Acetylcholine as a Neurotransmitter-60
The Nicotinic Acetylcholine Receptor-60
Neuromuscular Junction Structure and Transmitter
Release-60
Synaptic Vesicles: Transmitter Storage and Release-61
The Postsynaptic Response and Transmitter Disposal-63
Muscle Weakness due to Failure of Transmission at the
Neuromuscular Junction-63
Tetanus-63
The Inhibitory Cys-Loop Receptors-64
Serotonin (5-HT) and Emesis-64
Cys-Loop Receptors at Targets in Parasites-65
G Protein–Coupled Transmitters-65
Long-Term Potentiation by Phosphorylation-65
G Proteins Have Receptors as Exchange Factors-65
Glutamate and the G Proteins-66
Muscarinic Receptors-66
The Biogenic Amines Modulate Synaptic
Activity-66
Catechol Synthesis and Storage-67
Serotonin Synthesis and Functions-68
Catechol Release and Reuptake-69
Psychotropic Drugs and the Uptake and Delivery of
Catecholamines-69
G Protein Receptors for the Biogenic Amines-70
Purine Neurotransmission: Sensation and Coffee
Too-70
Peptides Modulate Brain Function-70
Retrograde Transmission-70
Coda-71
The adult brain averages 86 billion (∼1011) neurons plus an equal
number of nonneuronal support cells according to recent esti-
mates. Each neuron can make thousands of terminal contacts,
meaning that there are 1014 to 1015 connections within the
brain, with still more in the periphery. We can bring some order
to this massive system by understanding the chemistry used to
and directing the body’s activity.
THE SYNAPSE: ADAPTABLE TO MANY TASKS
The synapse is adaptable to many tasks. Controlled release of one
or more small molecules is the keystone of nerve communication,
and these small molecules are the nervous system’s neurotransmit-
ters, typically stored in small synaptic vesicles that are released at
synapses—specialized structures incorporating a presynaptic neuron
and a postsynaptic target cell (Fig. 4.1). The structure and function
of individual synapses are customized according to the requirements
of a particular task. The synapse must regulate the synthesis, stor-
age, and release of one or more transmitters, and signal transduction
must include one or more receptors at the target cell and a mecha-
nism to terminate the signal. This chapter will discuss the individual
classes of transmitters in some detail, using each to illustrate one or
more of these six principal features of chemical transmission.
Structure and Function
In many cases, neurotransmission occurs at a complex structure
comprising the nerve terminal, a synaptic space, the postsynaptic
cell, glia, and other supporting structures—the synapse. Individual
cases show considerable variability: The synapse may occur any-
where along the neuron’s axon. While the postsynaptic element is
generally a dendrite and an axon terminal is often the presynaptic
element, neuron cell bodies, axons, and dendrites can all be pre-
synaptic or postsynaptic in nature. Indeed, the synapse may release
a hormone that acts at another location entirely. Regardless, in each
case the elements are adapted to a specific purpose: to optimize
transmitter storage, its release, its effects, and its disposition. 
Transmitters and Modulators
The majority of vesicles in the nervous system contain one or the other
of the common transmitters (Table 4.1): glutamate, γ-aminobutyric
acid (GABA), glycine, or acetylcholine. Most synapses also use mod-
ulators of synaptic activity whose release is tightly regulated, such as
the biogenic amines (dopamine, norepinephrine, or serotonin) and
the neuropeptides. Still others, such as nitric oxide and the endor-
phins, diffuse from the neuron as they are synthesized. 
Synthesis and Storage
Most synapses synthesize multiple neurotransmitters from pre-
cursors that require specific transporters to cross the nerve’s
plasma membrane and often the blood-brain barrier. Synthesis
and storage in small synaptic vesicles is generally an ongoing pro-
cess in the nerve terminal, supplemented by neurotransmitter and
neurotransmitter precursors that are recycled from the synaptic
cleft and neighboring astrocytes (Fig. 4.1). Neurotransmitters are
often stored together, often with one molecule as the counter-
ion (adenosine triphosphate [ATP], glutamate) to another (ace-
tylcholine, the biogenic amines), to neutralize electrical charges
(Fig. 4.2). Solute and primary active transporters are adapted to
specific storage and uptake needs (Table 4.2). Neuropeptides are
a special case as they are synthesized in the soma and are stored
in dense-cored vesicles (DCVs) that package the peptide with
another major neurotransmitter. 

54
 Chemical Signaling in the Nervous System 55

Typical neuron to neuron synapse Glutaminergic synapse

Microtubule
mGluR2,3

Axon

Synaptic vesicles mGluR3

KA

Nerve
terminal Astrocyte

Active zone

AMPA
Synaptic Transporter
cleft

Plasma Enzyme
membrane EAAT1,2
Receptor
Postsynaptic AMPA NMDA EAAT3 mGluR1
neuron

Fig. 4.1  Delivery of the chemical signal at a synapse. Synapses are the chemically and anatomically spe-
cialized sites where neurons deliver chemical signals to target cells (left). In the typical neuron to neuron
synapse, anterograde axon transport delivers synaptic vesicles to the nerve terminals, where they advance
to the active zones to be primed for release. When an action potential depolarizes the terminal, voltage-
dependent calcium channels open, allowing calcium to enter. It is this calcium that triggers docking pro-
teins to fuse the vesicle membrane to the plasma membrane, releasing the neurotransmitter into the
synaptic cleft, where it binds to receptors on the target cell. The chemical signal is terminated when the
neurotransmitter is transported back up into the nerve terminal for reuse or changed into an inactive
metabolite by an enzyme on the postsynaptic membrane or in the synaptic cleft. The particular details of
synaptic function differ greatly from one site to the next, and the glutamatergic synapse (right) diverges
from the general schema in a variety of ways. For instance, multiple varieties of glutamate receptors (the
AMPA, NMDA, KA ionotropic receptors, and three mGluR metabotropic receptors) are found on the
target cell, on the presynaptic terminal and on neighboring astrocytes. Moreover, the cleft glutamate is
not degraded enzymatically or returned to the neuron where it originated, but it is removed by transport
proteins on the target cell (EAAT2) and surrounding astrocytes (EAAT1 and EAAT3) for later processing
(see Fig. 4.5). AMPA, AMPA receptor; EAAT, excitatory amino acid transporter; KA, kainate receptor;
mGluR, metabotropic glutamate receptor; NMDA, NMDA receptor.

Release
Release of the vesicle-stored transmitters is tightly regulated,
with synapses able to release variable numbers of vesicles accord-
ing to the strength of the signal to the nerve terminal. Vesicles
themselves may open transiently, releasing only part of their con-
tents, or they may fuse completely with the plasma membrane
of the nerve. Some DCVs are released constitutively, and their
transmitter’s release is controlled at the point of synthesis. Lipo-
philic neurotransmitters cannot be contained by vesicles, and so
they are released as soon as they are made.  by a nerve terminal is strictly limited to the immediate vicinity of
Transduction
All transmitters act by binding to some molecule, and that mol-
ecule is thus called its receptor. Occasionally, synapses consist of
a single kind of transmitter acting on a single kind of receptor,
resulting in a single kind of response, but most synapses con-
tain multiple receptor types and use more than one transmitter.
Receptors themselves can be simple, with a single binding site for
the neurotransmitter and a single function, but most exhibit exu-
berant extracellular and intracellular structures that are the result
of a wide variety of translational and posttranslational modifica-
tions and that are subject to allosteric modulation by metabolites,
drugs, and chemical modifications. The response can be excit-
atory or inhibitory, depending on the receptor (but not the trans-
mitter, because many transmitters can stimulate some processes
and inhibit others, even at the same synapse). The responses
can be divided into fast and slow processes, the fast synaptic
responses being due to a change in the open or closed state of an
ion channel (the ionotropic receptors) and the slow responses
(the metabotropic receptors) being due to a change in the state
of a G protein or some other enzyme (Table 4.3). 
Termination
For much of the nervous system, any transmitter that is released
the synapse, either by enzymatic digestion or by being recycled
back into the presynaptic terminal. Transmitter that escapes the
cleft is sequestered by solute transporters on the astrocytes or
the postsynaptic cell or is removed by the cerebrospinal fluid
or the vasculature (Table 4.2). In the special case of neurohor-
mones, release is designed to escape degradation because the tar-
get is distant. 
GLUTAMATE RECEPTORS: THE TETRAMERIC
EXCITATORY IONOTROPIC RECEPTORS
Glutamate is the neurotransmitter at 90% of the excitatory
synapses in the brain. Six families of glutamate receptors
exist in man: Three are ionotropic—the AMPA (α-amino-3-
hydroxy-5-methyl-4-isoxazolepropionic acid), the kainate, and
the NMDA (N-methyl-d-aspartate) receptors, and three are
56 Essential Concepts

metabotropic—group I, which increases inositol 1,4,5-trispho-

Table 4.1 Neurotransmitters
sphate (IP3) via Gq-coupled stimulation of phospholipase C and
MIMETICS AND groups II and III, which decrease cyclic adenosine monophos-
CLASS EXAMPLES ANTAGONISTS
phate (cAMP) via Gi/Go-coupled inhibition of adenylyl cyclase.
Gas NO, CO, H2S Nitroglycerine Glutamatergic synapses typically have ionotropic receptors
Ion Zn2+, Mg2+, Ca2+ on both the presynaptic and the postsynaptic membrane, with
Cholinergic Acetylcholine Succinylcholine, curare, metabotropic receptors off to the side plus accessory proteins
atropine, scopolamine that anchor the receptors and modify their activation and per-
Monoamine Epinephrine Yohimbine, propranolol, meability (Fig. 4.1). The synaptic glutamate is recovered via the
Norepinephrine and many antipsy- glutamate/glutamine cycle. First, glutamate is removed from
chotic drugs the synaptic cleft by transport into surrounding astrocyte (via
Dopamine l-DOPA, clonidine, EAAT1 and EAAT2 [see Fig. 4.5B later]; all transporter acronyms
amphetamine are explained in Table 4.2) or into the postsynaptic neuron (via
Serotonin Ergotamine, LSD EAAT3), where glutamine synthase generates glutamine, inci-
Histamine Antihistamines dentally consuming potentially toxic ammonia. This synaptically
inert glutamine is recycled back into the cleft by an electroneu-
Trace amine Tyramine,
β-phenylethylamine, tral Na+/H+ countertransporter (SNAT3). Glutamine uptake
amphetamine by the glutamatergic nerve is by Na+ cotransport (via SNAT8),
Amino Acid Glutamate followed by glutamate synthesis via phosphate-activated gluta-
minase. Vesicle uptake of this anionic neurotransmitter is by the
GABA (γ-aminobutyric acid) Benzodiazepine
vesicular glutamate transporter (VGLUT), driven by the inte-
Glycine Strychnine rior-positive electrical gradient (Fig. 4.2).
Purine Adenosine Caffeine
ATP (adenosine triphosphate) AMPA Receptors and Milliseconds of Excitation
Opioid Enkephalin Morphine AMPA and kainate receptors are the most common of the iono-
Eicosanoids Leukotriene B4, C4 Aspirin tropic glutamate receptors and open only briefly (for millisec-
Cannabinoid 2-Arachidonoylglycerol Tetrahydrocannabinol onds) before they become desensitized. AMPA receptors (with
Anandamide four possible subunits, GluA1 to GluA4) and kainate receptors
Peptide VIP (vasoactive intestinal (with five possible subunits, GluK1 to GluK5) have a similar
polypeptide) structure and function.
Np-Y (neuropeptide-Y) AMPA receptors are large—twice the size of the nicotinic
receptors and four times the size of the glutamate metabotropic
Angiotensin
receptors (Fig. 4.3). The outermost extracellular portion is the
The soluble endogenous ligands of the nervous system can be grouped into nine classes of N (amino)-terminal domain (ATD or NTD), and the portion
chemicals. Common examples are listed along with the more common naturally occurring ther- nearest the membrane is the ligand binding domain (LBD),
apeutic and toxic agonists or antagonists. A comprehensive source of information about iono-
tropic and metabotropic receptor proteins is found at http://www.guidetopharmacology.org. with the transmembrane domain (TMD) containing a box-like
CO, carbon monoxide; l-DOPA, l-3,4-dihydroxyphenylalanine; H2S, hydrogen sulfide; LSD, selectivity filter and gating apparatus. The AMPA ion pore is
lysergic acid diethylamide; NO, nitric oxide.
  
a highly conserved structure, present in potassium channels

Table 4.2  Membrane Transport Proteins of the Synapse

CLASS OF STOICHIOMETRY OF
TRANSPORTER GENE(S) PROTEIN(S) TRANSPORTED SUBSTANCE(S) FUNCTIONAL PURPOSE
Primary Active Transport
P-type 2 gene products Dimer of dimers 3 Na+ out/2 K+ in per ATP Establish sodium gradient
Na+-K+ ATPase
V-type 12 gene products 4 subunits, 23 2 H+ in per ATP Acidify cell organelles
H+ ATPase proteins
Anion Channel/Pump Transporters
CLC transporters CLCN-3 CLC-3 Chloride ↑ Chloride channel
and channels
CLCN-5 CLC-5 2 Chloride/proton ⇅ Vesicular chloride exchanger
Solute Carrier
MFS (Major Facilitator Superfamily) Clan
Glucose transporters SLC2A1 GLUT1 Glucose ↑ Transport of glucose across the
blood brain barrier
SLC2A3 GLUT3 Glucose ↑ High-affinity, non–insulin-
dependent glucose transport
into neurons
SLC2A4 GLUT4 Glucose ↑ Insulin-dependent glucose trans-
porter in fat and muscle
Monocarboxylic acid SLC16A1 MCT1 H+ and ketone body ⇈ Ferries ketone bodies across the
transporter blood-brain barrier
Organic anion SLC17A1, SLC17A2, SLC17A3 VGLUT1, 2 and 3 Glutamate ↑ Loads vesicles with glutamate
transporters
SLC17A9 VNUT ATP ↑ Loads vesicles with ATP
Continued
 Chemical Signaling in the Nervous System 57

Table 4.2  Membrane Transport Proteins of the Synapse­—cont’d

CLASS OF STOICHIOMETRY OF
TRANSPORTER GENE(S) PROTEIN(S) TRANSPORTED SUBSTANCE(S) FUNCTIONAL PURPOSE
Vesicular monoamine SLC18A1 VMAT1 Monoamine/proton ⇅ Loads vesicles with biogenic
transporters SLC18A2 VMAT2 amines
SLC18A3 VAChT Acetylcholine/proton ⇅ Loads vesicles with
Acetylcholine
Organic cation SLC22A3 EMT Uncertain Extraneuronal monoamine
transporter transporter
Equilibrative nucleo- SLC29A1, SLC29A2 ENT Nucleoside ↕ Reuptake of ATP
side transporters
SLC29A4 PMAT Monoamine ↕ Presynaptic reuptake of mono-
amines plus adenosine
APC (Amino Acid Polyamine Organocation) Clan
High-affinity choline SLC5A7 ChT Na+ and choline ⇈ Choline uptake into nerve
transporter terminal
Large neutral amino SLC7A5/SLC3A2 dimer LAT1 Phenylalanine/a.a. ⇅ Transport of phenylalanine,
acid transporter tyrosine/a.a. ⇅ tyrosine and l-DOPA across
l-DOPA/a.a. ⇅ the blood-brain barrier
Na+-dependent SLC38A1 SNAT1 Na+ and glutamine ⇈ “A” system uptake of glutamine
neutral amino acid SLC38A8 SNAT8 into soma and dendrites
transporters
SLC38A3 SNAT3 Na+ and glutamine ⇈/proton ⇅ “N” system astrocyte release of
glutamine
SDF (Sodium:Dicarboxylate Symporter Family) Clan
Excitatory amino SLC1A1 EAAT3 3Na+ and H+ and glutamate ⇈/K+ ⇅ Astrocyte uptake of glutamate
acid transporters SLC1A2 EAAT1
SLC1A3 EAAT2 3Na+ and H+ and glutamate ⇈/K+ ⇅ Postsynaptic uptake of
glutamate
SNF (Sodium:Neurotransmitter Symporter Family) Clan
Presynaptic catechol- SLC6A1 GAT1 3 Na+ and Cl− and GABA+ ⇈ Neuronal reuptake of GABA
amine re-uptake
SLC6A2 DAT 2 Na+ and Cl− and DA+ ⇈ Neuronal reuptake of dopamine
transporters
SLC6A3 NET Na+ and Cl− and NE+ ⇈ Neuronal reuptake of
norepinephrine
SLC6A4 SERT Na+ and Cl− and 5-HT+ ⇈ Neuronal reuptake of serotonin
SLC6A5 GlyT2 3 Na+ and Cl− and gly ⇈ High-affinity neuronal reuptake
of glycine
SLC6A9 GlyT1 2 Na+ and Cl− and gly ⇈ Low-affinity buffering of glycine
by astrocytes
SLC6A11 GAT3 3 Na+ and Cl− and GABA+ ⇈ GABA uptake by astrocytes

Transport of small molecules is crucial to the process of chemical signaling. (1) Primary active transport concentrates sodium and hydrogen ions via hydrolysis of ATP. (2) CLC proteins trans-
port chloride ions via pump or channel routes. (3) SLC proteins transport solutes according to the electrical and chemical gradients across cell and vesicular membranes. More 300 human SLC
proteins in 46 gene families have been identified, many of which are central to neuron function. Phylogenetic studies of gene homologies have grouped these gene families into 23 superfamilies,
4 of which are important to the mechanisms discussed in this chapter. A comprehensive source of information about transport proteins is found at: http://www.guidetopharmacology.org.
⇅, Countertransport; ⇈, cotransport; ↑, unidirectional transport; ↕, equilibrative transport; a.a., amino acid; ACh, acetylcholine; ATP, adenosine triphosphate; CLC, chloride channel; CLCN,
chloride voltage-gated channel; EMT, extraneuronal monoamine transporter; GABA, γ-aminobutyric acid; 5HT, serotonin or 5-hydroxytryptamine; SLC, solute carrier; SNAT, sodium-coupled
neutral amino acid transporter; VAChT, vesicular acetylcholine transporter; VGLUT, vesicular glutamate transporter; VMAT, vesicular monoamine transporter; VNUT, vesicular nucleotide
transporter.
  

throughout the tree of life, from bacteria (the Streptomyces
KcsA channel) to man (the Kv11.1 ether-a-go-go channels, one
of which repolarizes the cardiac action potential). If the GluA2
subunit is present, which is the usual case, the channel is per-
meable to sodium and potassium ions only. RNA editing causes
the transcribed sequence for glutamine at codon 586 to be
translated as an arginine in GluA2. This arginine adds an addi-
tional positive charge to the selectivity filter of the ion pore,
thus excluding divalent cations such as calcium. Codon 586 is
not A→I RNA edited in the other subunits, and so any AMPA
receptor made without GluA2 is permeant to calcium. These
calcium-permeant AMPA channels have a conductance that
declines as the membrane is depolarized (an inward rectifica-
tion) due to block of the ion pore by endogenous cytoplasmic
polyamines. This inward rectification is of some benefit for a
calcium permeant channel because it will lessen the quantity of
calcium that enters the cell, reducing the toxic effects of a high
cell calcium and minimizing the energy required to pump the
calcium back out.
Agonists bind to a clamshell slot in the LBD, which then snaps
shut behind. With two glutamates bound, the channel begins to
flicker brief openings. When a third glutamate binds, the chan-
nel opens for longer, giving more current, and with a fourth glu-
tamate, the channel is open most of the time with still more
current flowing. However, within seconds, the receptor becomes
desensitized as dimers separate in the LBD and channel activity
ceases.
The duration of the glutamate response at the AMPA recep-
tor can be prolonged in a variety of ways. Alternative splic-
ing occurs in a TMD exon, leading to slower AMPA closing
in the fetal form than the adult variant. Phosphorylation of
two serines by protein kinase C (PKC) and another by protein
58 Essential Concepts

Table 4.3  Receptors in the Nervous System

CLASS SUBFAMILY EXAMPLES
A A17 Adrenergic receptors (α1A, 1B, 1D, 2A, 2B, 2C and β1, 2, 3)
(19 subfamilies) Dopaminergic receptors (D1, 2, 3, 4, 5)
Histaminergic receptor (H2)
Serotonergic receptors (5-HT2A, 2B, 2C,6)
A18 Adenosine (previously P1) receptors (A1, 2a, 2b, 3)
Histaminergic receptors (H1, 3, 4)
Muscarinic cholinergic receptors (M1, 2, 3, 4, 5)
A11, A12 Purinergic receptors (P2Y1, 2, 4, 6, 8, 11, 12, 13)
A19 Serotonergic receptors (5-HT1A, 1B, 1D, 1E, 1F, 4, 5A, 7)
A4 Opioid receptors
A13 Cannabinoid receptors
A16 Cone pigments that transduce light
Unclassified Olfactory and vomeronasal receptors
B B1 Vasoactive intestinal peptide (VIP) receptor
(3 subfamilies) Calcitonin, glucagon, and secretin hormone receptors
C C2 GABAB receptor
(5 subfamilies)
C3 Glutamate receptor (mGluR1, 2, 3, 4, 5, 6, 7, 8)
C5 Taste receptors

All G protein–coupled (GPC) receptors contain seven transmembrane α helices. Approximately 800 genes in the human genome are predicted to be GPC member proteins, and they are
divided into three classes, according to structural homology; each of the classes is further divided into numerous subfamilies whose members subsume a wide variety of functions throughout
the body. Examples relevant to the nervous system are listed here, but a comprehensive source of information about G protein receptors is found at http://www.guidetopharmacology.org.
GABA, γ-aminobutyric acid; 5-HT, 5-hydroxytryptamine.
  

ATP

V-ATPase

Cytoplasm ADP + Pi

2H+
Cl–
CLC-3 VGLUT
CLC-5 VMAT

∆Ψ = 80 mv

2Cl– H+ ∆pH = 1.5 Dopamine

Synaptic Glutamate
vesicle
interior
Norepinephrine Dopamine β-hydroxylase

Fig. 4.2 Filling the synaptic vesicle: norepinephrine as an example. Synaptic vesicles concentrate
their neurotransmitters by electrical and proton gradients generated by the vacuolar-type H+ ATPase
(V-ATPase). Like the mitochondrial F1FoATP synthase (but rotating in the opposite direction), V-ATPase
catalyzes the active transport of protons into the vesicle via a rotating subunit powered by ATP hydrolysis.
The V-ATPase protein has four components: a catalytic subunit that is attached to the actin cytoskeleton,
a central stalk, a rotating subunit in the plane of the membrane’s bilayer and a peripheral stalk. Hydrolysis
of ATP in the catalytic subunit turns the rotor in a clockwise direction at speeds of up to 103 rpm, import-
ing protons through an H+ channel between the peripheral stalk and the rotor. One complete rotation
translocates three protons, one of which is consumed by a cycle of the CLC-5 2-chloride/1-proton coun-
tertransporter; the net result is a gain of two H+Cl−. This schema preserves microscopic electoneutrality,
ultimately allowing the hydrogen ions to become 30-fold concentrated relative to the cytoplasm, a ΔpH
of 1.5. Passive efflux of the chloride ion through the CLC-3 channel generates a diffusion membrane
potential (Δψ), with the vesicle being inside 80-mV positive. The proton gradient drives countertransport
uptake of dopamine, norepinephrine, serotonin, and acetylcholine by vesicular monoamine transporters
such as VMAT. Uniquely among the small neurotransmitters, the final step of norepinephrine synthesis
occurs within the synaptic vesicle, where dopamine β-hydroxylase adds a hydroxyl group to dopamine.
The membrane potential drives uptake of the negatively charged neurotransmitters glutamate and ATP via
organic anion transporters such as VGLUT. Thus, in this example, the synaptic vesicle will contain two
neurotransmitters, the cation norepinephrine and the anion glutamate. ADP, adenosine diphosphate; ATP,
adenosine triphosphate; CLC, chloride channel; VGLUT, vesicular glutamate transporter; VMAT, vesicular
monoamine transporter.

ATD
LBD
TMD
Fig. 4.3 The glutamate ionotropic receptor. The homotetrameric GluA2
AMPA receptor is shown as viewed from the front (left) and from the side
(right). AMPA receptors have four subunits that associate as pairs of dimers,
making a lentoid extracellular component that is wider than deep in this N
assembly; heterotetramers can also exist as an O assembly, which is more com-
pact. The outermost portion of the receptor is the N-(amino) terminal domain
(ATD or NTD) and the portion nearest the membrane is the ligand binding
domain (LBD, separated by the thin line). The ATD is a pair of dimers, as the
transmembrane domain (TMD), but with partners swapped. That is to say, for
a receptor made up with two GluA2s, one GluA1 and one GluA3, the ATD
might pair a GluA2-GluA1 dimer with a GluA2*-GluA3 dimer, but then the
LBD would pair GluA2-GluA3 with GluA2*-GluA1, thus tightly binding the
heterotetramer into a functional whole. This twofold symmetry transforms into
a fourfold symmetry at the TMD (the width of the cell membrane marked
in gray) to contain the box-like selectivity filter and gating apparatus. The
TMD houses the pore, whereas the extracellular domain contains the ligand
binding site. The NMDA ATD domain is larger and more compact, render-
ing it more oblate than lentoid. As a consequence, more of its sequence is in
contact with the other GluN subunits and the LBD domain, which leads to
more control of the receptor function. AMPA, α-amino-3-hydroxy-5-methyl-4-
isoxazolepropionic acid; NMDA, N-methyl-d-aspartate.
kinase A (PKA) increases and prolongs the glutamate response.
Transmembrane AMPA receptor proteins (TARPs) modify the
lifetime of the glutamate response by binding to the TMD. For
instance, stargazin increases the rate of channel opening and
slows desensitization.  but activating calcium impermeant receptors may result in a pro-
NMDA Receptors and Seconds of Activation
NMDA receptors open to allow a large influx of calcium, but
only after the receptor’s cell has been depolarized repeatedly
over a period of hundreds of milliseconds. This calcium modu-
lates many and varied cell processes by activating Ca2+/calmodu-
lin-dependent kinases, protein kinase C, phospholipase C and A2,
calcineurin, and nitric oxide synthase. In cases of large, prolonged
NMDA stimuli, this calcium signal is great enough to damage the
cell and even be fatal. Functionally, the duration and amplitude of
the NMDA response is modulated by many ions, by endogenous
molecules, and by psychotropic drugs. It has been tentatively
implicated in Alzheimer and Parkinson diseases, depression,
schizophrenia, and ischemic neuron death after a stroke.
NMDA receptors are obligate heterodimers, requiring a pair of
GluN1 to GluN2 dimers. There is only one gene encoding GluN1
(GRIN1), but it is alternatively spliced into eight variants. Four
GluN2 monomers subtypes exist, and they are distributed with
regional specificity throughout the brain. All monomers bind one
glutamate, but only after the two GluN1s have bound a differ-
ent ligand: d-serine. Glia synthesize d-serine (one of the only
two d-amino acids to be used by vertebrates) from l-serine via
serine racemase and release it into the synaptic area. Glycine, the
inhibitory neurotransmitter of the brainstem and spinal cord, can
substitute for d-serine.
Chemical Signaling in the Nervous System 59
The ATD of the NMDA (but not AMPA or kainate) recep-
tor senses its surroundings’ pH, opening less frequently and for
briefer periods as the receptor’s extracellular surroundings are
made more acidic, potentially protecting the brain during peri-
ods of ischemic acidosis. Polyamines inhibit this effect. Little
sequence homology is shared between the NMDA and AMPA/
kainate ATDs.
The requirement that a cell be depolarized for an extended
period of time before its NMDA receptors appear to open is
explained by the presence of twin asparagines at the selectivity
filter of the NMDA (but not the AMPA/kainate receptor) chan-
nel. Extracellular magnesium binds to the electronegative oxy-
gens of these twin asparagines and is released only slowly when
the membrane is depolarized from rest, a change in voltage that
provides the electrical force to drive the divalent ion out of the
channel. Thus the NMDA receptor will allow no current to pass
even when activated by agonist at the resting potential because
it is blocked by a magnesium ion. It is only with repeated glu-
tamate stimuli acting on neighboring AMPA receptors that the
membrane potential is near zero for long enough for blocking
magnesium ion to exit the pore, allowing measurable currents
to flow through a channel that was otherwise open the whole
time. Limiting calcium entry is particularly important in the
case of the NMDA receptor, both because its calcium currents
are large when the magnesium block is finally relieved, but also
because the receptors are often found in the membrane of den-
dritic spines, where the volume of the terminal bud is small
and the cytoplasm is separated from the nerve cell body by a
narrow stalk that restricts diffusion into and out of the spine. In
these spines, any calcium entry will have a disproportionately
large effect. 
Kainate Receptors: AMPA-like, but Presynaptic
The kainate receptors are predominantly, but not exclusively,
presynaptic. Sharing the structure and function of AMPA
receptors, kainate receptors are generally permeant only to
sodium and potassium because RNA editing has resulted in the
glutamine→arginine substitution at codon 586, but may be cal-
cium permeant because this editing is not obligatory. The cal-
cium permeant kainate receptors augment transmitter release,
longed depolarization that is inhibitory at certain synapses and
excitatory at others.
Five kainate monomers exist that can combine as homotetra-
mers or heterotetramers. GluK1, GluK2, and GluK3 combine
to form channels, whereas GluK4 and GluK5 do not, merely
being able to modify the function of the heterotetramer they
participate in. The structure of the kainate receptor’s LBD
allows domoic acid to bind tightly to the ligand site, causing
a prolonged activation. This toxin is produced by a red algae,
and it is concentrated by mollusks during a bloom of that algae,
resulting in sporadic cases of the amnestic shellfish poisoning
syndrome. Eating tainted shellfish can result in neurotoxic lev-
els of calcium influx through kainate receptors in the hippo-
campus and amygdala. 
CYS-LOOP RECEPTORS: THE PENTAMERIC
MULTIFUNCTIONAL IONOTROPIC RECEPTORS
Cys-loop receptors are pentamers of four or five different mono-
mers that might have acetylcholine, GABA, glycine or serotonin
as its agonist. The structure and function of the nicotinic acetyl-
choline receptor (nAChR) was discussed at length in Chapter
3. This section will present details about the structure of the
synapse, about how the transmitter is stored and released, how
failures in synaptic function occur, and, finally, how other trans-
mitters use this receptor type.
60 Essential Concepts

Acetylcholine as a Neurotransmitter
Acetylcholine activates receptors that are ionotropic (the
nAChR) and metabotropic (the muscarinic acetylcholine recep-
tor, mAChR), acting in the central nervous system (CNS) (e.g.,
to increase dopamine (DA) via the mesolimbic pathway), the
periphery (the neuromuscular junction [NMJ]), and in the
autonomic nervous system (the preganglionic neurons act on
nicotinic receptors, whereas postganglionic parasympathetic
neurons act on muscarinic receptors). We will next focus on the
NMJ, which has long been the subject of experimental investiga-
tions because it is so readily accessible.  Release
The Nicotinic Acetylcholine Receptor
The ion channel of the cys-loop transmitter-activated receptors
was introduced by examining the nAChR of the NMJ in Chapter
3. Patch clamp recordings (see Fig. 3-9) of this channel showed
patterns of ion channel openings and closings that reflected the
action of agonists and explained conceptually how partial ago-
nists, antagonists, and depolarizing blockers might arise. The
molecular structure (see Fig. 3-8) demonstrated sites for ago-
nist binding and ion permeation. Signaling by ACh at the NMJ
requires a highly specialized synapse, whose structure and func-
tion is optimized for motor activity, but whose constituents are
the site of much pathology and dysfunction. 
Neuromuscular Junction Structure and Transmitter
Motor nerve axons branch at their termini to innervate many
muscle fibers, all of which contract together as a motor unit.
The myelin sheath stops short of the synapse, exposing the
naked axon to the muscle membrane at a specialized disk-
shaped region called the motor end plate (Fig. 4.4). Numerous

Schwann cell
Motor end plate
Synaptic vesicles at dense bars
Axon terminal
Motor Basal lamina
fiber
Nucleus of
muscle cell
Mitochondria
Synaptic cleft
Subjunctional folds

Myofibrils

Dense bar/active zone

Synaptic vesicle
Presynaptic
membrane

Calcium channels
Basal lamina Synaptic cleft

Postsynaptic
Acetylcholine
membrane
receptors

Subjunctional fold
Acetylcholinesterase

Sodium channel

B
Fig. 4.4  The neuromuscular junction. (A) Motor neurons end as a plate-appearing terminus that is a
highly specialized synapse to the skeletal muscle. The neuron’s terminal organizes clusters of synap-
tic vesicles at the active zones in preparation for a large release of neurotransmitter quanta. Release
occurs when an action potential depolarizes the membrane and opens voltage-dependent calcium
channels that deliver the calcium signal to release the vesicles (B). The folds of the postsynaptic mem-
brane closest to the nerve terminal contain receptors that recognize the newly released neurotrans-
mitter, opening nonspecific cation-permeant channels that initiate an action potential in the skeletal
muscle. The stimulus is rapidly terminated when the neurotransmitter diffuses into the synaptic folds
away from the receptors and toward acetylcholinesterase molecules that dispose of the transmitter
by hydrolysis.
 Chemical Signaling in the Nervous System 61

transmitter-filled vesicles fill the nerve terminal, both large
and small. The large vesicles are the DCVs, whereas the more
numerous small vesicles (SV) are clear and are concentrated at
the active zones, where dense bars of docking proteins cluster
in the plasma membrane with voltage-sensitive calcium chan-
nels nearby. Two docking proteins (SNAP-25 and syntaxin) are
preassembled at the dense bars and are chaperoned by Munc
proteins so that additional unwanted assemblies are prevented.
Two proteins in the synaptic vesicle membrane complete the
vesicle’s docking: a synaptotagmin, which senses increases in
cytoplasmic calcium, and synaptobrevin, the third and final
docking protein, which displaces the Munc proteins and forms
a tight assembly with the SNAP-25/syntaxin dimer. The three
docking proteins together form the v/t SNARE complex,
which firmly anchors the membrane of the synaptic vesicle
to the nerve terminal’s plasma membrane, allowing them to
fuse and the vesicle’s contents to exit when an AP increases
cytoplasm’s calcium concentration. The vesicle’s membrane is
quickly retrieved by clathrin-mediated endocytosis once NSF,
an AAA+ ATPase, causes the ultrastable SNARE complex
to dissociate. These newly formed (but empty) vesicles then
return to the synaptic store.
Not all docking encounters proceed to completion. First,
transient Ω-shaped encounters result in a flickering fusion
pore that extends from the vesicle interior to the extracellular
space through a short stalk. At this point, the fusion pore is
∼1 nm in diameter with a conductance of ∼80 ps, placing it
within the realm of the large nonspecific cation channels. Even
at this size, the small neurotransmitters such as acetylcholine
can exit. Although this stage may rapidly proceed to a full
fusion and complete release of vesicle contents, the encounter
may break off prematurely if the fusion pore closes before full
fusion (yclept “kiss-and-run”). 
Synaptic Vesicles: Transmitter Storage and Release
The NMJ stores its ACh in small, ∼40 nm, lucent synaptic ves-
icles that are much like the vesicles that other nerve terminals
use to store their glutamate, GABA, glycine, or other small neu-
rotransmitters (Fig. 4.5). As with mitochondria and many other
organelles descended from gram-negative bacteria, synaptic ves-
icles processes are driven by a proton gradient, in this case with
the vesicle interior being acidic and electrically positive. In all
synaptic vesicles, the proton gradient is established by the vacuo-
lar-type H+-ATPase (Fig. 4.2). This proton gradient is harnessed
by H+ countertransporters to concentrate positively charged
neurotransmitters such as acetylcholine and the biogenic amines
and uncharged neurotransmitters such as GABA. The positive
electrical gradient is used to concentrate glutamate, an anion.
In each case, filling such synaptic vesicles requires that the neu-
rotransmitter be first synthesized and then transported into the
membrane-bound space that is the vesicle.
ACh synthesis (Fig. 4.6) is limited by the availability of cho-
line, its precursor. Choline cannot be synthesized by the neuron,
but instead is transported into the terminal by the high-affinity
choline transporter (ChT, Table 4.2). To match the supply of
this neurotransmitter with its demand, more than 90% of the
neurons’ ChT is in the synaptic vesicle’s membrane; exocyto-
sis inserts ChT into the plasma membrane transiently, until the

Typical neuron to neuron synapse Glutaminergic synapse

Axon
SNAT3 Astrocyte
SNAT8

Synaptic vesicles Glutamine

Gln

Nerve Glu
terminal
Active zone
Clathrin
Clathrin

Synaptic Transporter
cleft Glutamate
EAAT1,2
Plasma
membrane

Postsynaptic EAAT3
neuron

Fig. 4.5  The dynamics of small, clear synaptic vesicles. The various membrane proteins found in the
small, clear synaptic vesicles are packaged in the endoplasmic reticulum and then the Golgi apparatus and
delivered to the nerve terminal by axon transport. A fraction of the synaptic vesicles cluster at the active
zone, where an action potential will cause their exocytosis. The membrane is later retrieved by clathrin-
coated endocytosis, which also retrieves the membrane proteins that had been present in the vesicle (left).
In many synapses, the neurotransmitter is retrieved from the synaptic cleft, and the vesicle is then refilled
and recycled into the synaptic pool (right). For glutamate and GABA, the neurotransmitter is metabolized
to an inactive amine within neighboring astrocytes and then returned to the synaptic cleft. Like choline,
this inactive metabolite must be returned to the nerve terminal for the neurotransmitter to be resynthe-
sized and returned to the synaptic vesicle. EAAT, excitatory amino acid transporter; GABA; γ-aminobutyric
acid; GLN, glutamine; GLU, glutamate; SNAT, sodium-coupled neutral amino acid transporter.
62 Essential Concepts

OH
+
N
Choline

O
+
N
Acetylcholine
O O

O N
+ +
N O
Succinylcholine
O
Fig. 4.6  The acetylcholine family of chemical structures. Acetylcholine is synthesized by the enzyme
choline acetyltransferase (ChAT) from choline and acetyl-coenzymeA. Succinylcholine is a dimer of ace-
tylcholine, head to head, and is a paralytic drug because it acts by first activating nicotinic receptors, then
causing profound desensitization of the receptor. The modified groups are highlighted for emphasis.

vesicular membrane is reclaimed by endocytosis. Blocking ChT Endoplasmic

by hemicholinium-3 inhibits all cholinergic transmission because
the recycled cholinergic vesicles can no longer be refilled once
this ACh precursor is exhausted. In practice, sympathetic and
parasympathetic functions are more impaired when choline
uptake is blocked than are motor functions.
To be effective, neurotransmitters must be highly concen-
trated in their vesicles, and ACh is dramatically concentrated—
to ∼0.1M—within the NMJ’s vesicles by VAChT, an H+-solute
countertransporter that is driven by the proton gradient across
the vesicular membrane.
Such an acidic interior has the incidental benefit of stabiliz-
ing ACh molecules because they hydrolyze back to choline and
acetate at neutral pH.
Synaptic vesicles require negatively charged counterions
because ACh has a positively charged quaternary nitrogen moiety
in its structure. Being a trivalent anion, ATP effectively provides
this negative charge and is transported into the vesicle by VNUT
(Table 4.2). Glutamate and phosphate anions are likely present
also because their transporter (VGLUT1) is also present in the
vesicle’s membrane.
Motor nerve terminals also contain larger (90-250 nm)
hybrid synaptic vesicles (the DCVs) that combine the molecu-
lar components of the small, clear synaptic vesicles with those
of Golgi-derived secretory granules to store both a small neu-
rotransmitter plus one or more peptides (Fig. 4.7). DCVs appear
electron dense because they contain a very high concentration
of protein: 300 mg/mL or more of granins, peptide precursors,
and assorted enzymes. The DCVs of even a single synapse are
heterogeneous in their contents; for instance, both substance
P and calcitonin gene–related peptide are found at the NMJ.
Once DCVs bud off the trans-face of the Golgi apparatus, they
undergo an hour-long maturation process, whereby the prohor-
mones are cleaved to size, the vesicles become competent to
undergo secretion as regulated by increased cell calcium, and
they are transported to their release sites, where they load up
on their small neurotransmitter.
Even in a single nerve terminal, release of DCVs exhibits het-
erogeneity in response to different patterns of stimuli, as well as
variations in the contents released and how the vesicles are dis-
posed of. The form of synaptotagmin (SYT) found on the DCV
membrane confers specific calcium sensitivities and release char-
acteristics. For instance, SYT7 responds to the calcium of a single
spike, with a high likelihood to empty the DCVs contents com-
pletely. When recovered from the plasma membrane, this SYT7-
containing DCV can refill with small neurotransmitters, but
the peptide cannot be replaced. The vesicle must return to the
Nucleus reticulum:
of Prepropeptide
neuron synthesis
Propeptide
Cis face
Golgi apparatus
Trans
face Maturing dense-cored
vesicle
Maturing synaptic vesicle
Anterograde transport
along axon
Fig. 4.7  Synaptic vesicle formation. Both the small clear synaptic vesicles (SV)
and the dense-cored vesicles (DCVs) bud off the trans face of the Golgi appara-
tus with membrane that contains the channels and transporter proteins appropri-
ate for that vesicle’s function. In addition, granin proteins are targeted to the
DCVs to assist in packaging the propeptides that had been synthesized in the
endoplasmic reticulum and processed in the Golgi stack. The DCVs will be fur-
ther processed as they are transported down microtubules to the nerve terminals,
with propeptides being cleaved into neuropeptide transmitters. Although the
SVs and DCVs can replenish their stores of small neurotransmitters during their
lifetime, the DCV peptide contents are loaded only at the moment when those
vesicles bud off the Golgi apparatus.
neuron’s soma for lysosomal disposal. At the other extreme, SYT1
requires much higher calcium concentration, so it will respond
only to a train of APs; unlike SYT7, SYT1 is a synaptotagmin
that likely results in a kiss-and-run transient fusion pore, which
releases the small neurotransmitter, but is too narrow to permit
the peptide to exit. After the fusion pore transient, this DCV can
refill with small neurotransmitters and return to the synaptic ves-
icle pool for a later release event that could be transient or com-
plete. Other synaptotagmins show intermediate behavior. Hence
a synapse’s DCV can be designed to release small and neuropep-
tide transmitters promptly at the first stimulus, to release only
the small neurotransmitters, or to conserve its peptide, releasing
it only occasionally in the face of a prolonged stimulus. 

The Postsynaptic Response and Transmitter Disposal
The membrane of the skeletal muscle fiber is thrown up into
numerous subjunctional folds in the end plate region, with the
tops of the folds situated immediately opposite the active zones
of the nerve terminal. Clustered on the top of the folds are
nAChRs, which respond to released neurotransmitter (acetyl-
choline) by increasing the conductance of the muscle membrane
to sodium and potassium. The released transmitter reaches the
nAChRs within tens of microseconds after release, ensuring a
speedy transmission. Equally important, the transmitter will
not have time to disperse, guaranteeing that the dense cluster
of nAChRs will be exposed to a high concentration of transmit-
ter; consequently, almost all the released transmitter will be
bound to and act on the receptors. Clustered at the base of the
folds are large numbers of voltage-dependent sodium channels,
guaranteeing that the muscle will respond with one AP for each
AP in the motor nerve.
Between the nerve and the muscle lies a synaptic cleft that is
narrow but very deep. The cleft matrix contains a basal lamina
with collagen and laminin that combine to keep the nerve and
muscle in close approximation and precise register during the
active muscle activity. The depths of the synaptic cleft contain
a high concentration of acetylcholinesterase (AChE) molecules
that are held in place by their long collagen-like tail. Thus one
purpose of the synaptic cleft is to trap ACh and to rapidly hydro-
lyze the transmitter into choline and acetate. This positioning
ensures that little ACh diffuses out from under the nerve termi-
nal and that the transmitter is present for only a brief (∼1 ms)
time.  desensitization progresses too far, the loss of functional AChRs
Muscle Weakness due to Failure of Transmission at
the Neuromuscular Junction
Muscle weakness can be the result of a failure at the NMJ.
Although the output of many synapses is proportional to their
inputs, the NMJ is designed to reliably trigger a muscle contrac-
tion each time the motor neuron fires. Hence agents that cause
muscle weakness via the NMJ must have a specific NMJ target
and be potent in their action. For instance, alcohols and local
anesthetics inhibit ACh by preventing ion permeation through
nAChRs; the vestibule of this receptor’s channel is large enough
to admit local anesthetic molecules such as lidocaine, which
bind tightly and prevent ions from passing. The region behind
the gating α-helix is also water filled and contains a specific site
for alcohol and the binding of local anesthetics as well. How-
ever, the low concentrations of alcohols and local anesthet-
ics that are achieved with recreational use or antiarrhythmic
therapy act only at the proportional synapses, not at the more
robust NMJ.
Toxins exist that act more specifically at the NMJ; hence
they effectively interfere with synaptic transmission at the
NMJ. The three botulinum toxins are metalloproteinases that
specifically attack the docking proteins syntaxin, synapto-
brevin, and SNAP-25 on the presynaptic side of the NMJ. This
effectively interrupts exocytosis and causes a paralysis that
lasts until new docking proteins are synthesized. α-Latrotoxin,
the toxin of the black widow spider, specifically binds to
neurexin, a membrane protein that participates in connections
between presynaptic and postsynaptic membranes. Tetramers
of α-latrotoxin insert into the nerve terminal’s membrane and
form nonspecific cation pores that flood the cytoplasm with
calcium, causing a massive emptying of the nerve terminal of
neurotransmitter vesicles and then a complete loss of that end
plate’s function.
Acquired myasthenias—those diseases characterized by mus-
cle weakness—can be due to the activity of the immune system.
Myasthenia gravis is the most common syndrome, character-
ized by the fluctuating severity of the weakness, by the early
Chemical Signaling in the Nervous System 63
involvement of ocular muscles, and by its response to choliner-
gic drugs. In myasthenia gravis, a small area of the extracellular
region of the AChR—nanometers away from the transmitter
binding site—is vulnerable to autoimmune attack. These amino
acids form an epitope that is shared by peptides expressed during
certain viral illnesses and by cells in the thymus that can activate
antigen-specific T lymphocytes. Antibodies generated as a con-
sequence of this activity may cross-link AChRs, increasing their
rate of endocytosis and subsequent lysosomal destruction, and
reducing their normal lifetime from a week to half that value.
When the increased rate of loss becomes sufficient, the current
generated during the end plate potential will be insufficient to
trigger the nerve AP, and muscle weakness will ensue. In other
individuals, the antibodies bind complement, leading to the for-
mation of membrane attack complexes (see Fig. 3-5) and lysis of
the muscle cell. In still other individuals, the antibodies are of
no pathologic consequence whatsoever, so a simple determina-
tion of antibody titer is not absolutely predictive of the severity
of the disease. The most direct treatment is the use of drugs
that inhibit cholinesterase activity, such as the anticholinesterase
pyridostigmine. In the longer term, the severity of the immune
response is reduced by the use immunosuppressive agents (for
instance, corticosteroids or more powerful immunosuppressant
therapy, or by removing the thymus.
Anticholinesterase therapy is designed to increase and to
prolong the synaptic concentrations of ACh. An untoward con-
sequence of intensive anticholinesterase therapy is the desensiti-
zation of AChRs by the prolonged exposure to ACh. When the
overtakes the benefit of the prolonged exposure to transmitter,
and the patient becomes weaker. This condition, termed a cho-
linergic crisis, is a therapeutic dilemma; any individual patient
might become weaker because the immune disease worsens or
because the number of AChRs is being reduced through desen-
sitization. The test to distinguish between the two possibilities
is the double-blind use of a short-acting cholinesterase inhibitor
(with a ventilator nearby in case the patient becomes too weak to
breathe). If the patient’s strength is improved, then the immune
disease is becoming worse, and the anticholinesterase therapy
must be increased; but if the patient weakens, then he or she is
in a cholinergic crisis, and the anticholinesterase therapy must be
reduced.
Muscle weakness may also accompany small cell carcinoma
of the lung owing to the production of antibodies to calcium
channels in the presynaptic nerve terminal. In this situation, the
weakness is a paraneoplastic process—a syndrome that is asso-
ciated with a primary neoplasm elsewhere in the body. Such a
condition is variously called the myasthenic syndrome or the
Lambert-Eaton syndrome. Characteristically, these patients
gain strength with repeated muscle activity, unlike those with
myasthenia gravis, who fatigue more rapidly than normal.
This improvement is due to repeated firing of the nerve AP
that causes potentiation or facilitation of transmitter release as
more calcium enters the nerve terminal with each succeeding
AP. With sufficient activity, many of the affected nerve termi-
nals will again have adequate calcium to release transmitter to
generate a muscle AP. 
Tetanus
Tetanus is a disease that is caused by a mechanism similar to bot-
ulism, although it operates on different neurons. Like botulinum
toxin, tetanus toxin is a bacterial product (from Clostridium tet-
ani), which has a specific binding affinity for nerve terminals at
the neuromuscular junction and has enzymatic activity specific
for a synaptic vesicle protein (synaptobrevin in this case). The
difference is that after uptake, the toxin is retrogradely trans-
ported back up the axon to the soma, where it is released into
64 Essential Concepts
Fig. 4.8  Opisthotonus. Tetanus leads to spasm of all muscles, so the strongest
will prevail, leading to an open-eye stare, extension of the back, flexion, and
inward rotation of the arms and hands. The condition was described and accu-
rately illustrated in 1809 by Charles Bell. Today, opisthotonus is seen in neonatal
tetanus, a fatal disease of newborn infants whose umbilical stump has become
infected and who lack passive immunity because their mother had not been
immunized against tetanus.
the extracellular space of the spinal cord and taken up by neigh-
boring nerve terminals. The predominant nerve terminals on the
soma belong to inhibitory interneurons, and it is here that the
toxic proteolysis actually occurs. When this interneuron becomes
unable to release its transmitter, its inhibitory input onto the
motor neuron is interrupted, leading to extreme hyperactivity in
the motor neuron and spasticity of the associated muscles. The
first muscle groups involved are those with short axons (face,
jaw, and neck, yielding the typical presentation of risus sardoni-
cus), and only later do muscle groups with longer axons become
involved (trunk, arms, and legs, leading to the characteristic opis-
thotonus posture, Fig. 4.8). Parasympathetic and preganglionic
sympathetic nerves are also targeted, leading to clinical features
of sympathetic overactivity. Residual muscle weakness is due to
the fact that the toxin can also be active at the neuromuscular
junction itself, an activity that is hidden by the initial, and far
more dramatic, spasticity.  tive, negative, or neutral effects. Positive allosteric modulators
The Inhibitory Cys-Loop Receptors
Glycine and GABA activate cys-loop receptors that inhibit nerve
activity. Glycine is the usual inhibitory transmitter at spinal cord
and lower brainstem synapses. It is generally released at well-
defined synapses onto another neuron’s cell body (axosomatic
synapses) or dendritic nerve terminals (axodendritic synapses).
GABA is the most common inhibitory transmitter in the rest of
the brain and acts at both ionotropic (GABAA) and metabotropic
(GABAB, Gi/o-coupled) receptors. There are two populations of
GABAergic neurons. Those with long axons release their trans-
mitter at conventional axosomatic and axodendritic synapses. A
second population of local circuit interneurons has short axons—
or no axon at all—and these cells can release transmitter at poorly
differentiated dendrodendritic synapses.
GABA sequestration is by astrocytes using the glutamate/glu-
tamine cycle (Fig. 4.5), except that GABA is first transaminated
to form glutamate to initiate the cycle. GABA is then recon-
stituted in its nerve terminal by decarboxylation of glutamate.
Vesicle uptake of this electrically neutral neurotransmitter is by
VGAT, an H+ countertransport mechanism driven by the proton
gradient of the synaptic vesicle.
Glycine is the neurotransmitter of the inhibitory postsynap-
tic potentials (IPSPs) generated by Renshaw cells and other
interneurons in the spinal cord, and it is cleared from the syn-
aptic cleft by high-affinity GlyT2 nerve terminal transporters
and is buffered by lower affinity astrocyte GlyT1 transport-
ers. Glycine opens chloride-selective ion channels, reducing
the membrane impedance and hyperpolarizing the cell body.
Consequently, there is a smaller probability that an AP will
be initiated at the axon hillock. This is because any excitatory
stimuli to the cell will be less effective at changing the mem-
brane potential and they will have to depolarize the membrane
further to initiate an AP.
The adult glycine receptor is typically a heteropentamer of
three α and two β subunits. There are four α and one β subunits,
with α2 being the subunit expressed by adults and α1 by embryos.
β Subunits anchor the receptor to the cytoskeleton at the active
zones of the nerve terminal. Agonists bind at the usual cys-loop
ligand binding site located between α subunits, as does strych-
nine, a receptor antagonist that locks the channel closed to cause
a tetanus-like hyperreflexivity and spasticity. In smaller doses,
strychnine has been used to improve strength and endurance in
school and competition, but the therapeutic window is narrow.
Ionotropic GABAA receptors exist in many forms in the brain,
retina, and periphery, with 19 isoforms in 8 families: α (6), β (3),
ρ (3), γ (3), δ (1), ε (1), θ (1), π (1). The most common pen-
tamer is α1β2γ2, with various permutations appearing in different
regions of the brain or even in different regions of one particular
cell, each with a particular pharmacologic profile. Of the α sub-
units, α1 is associated with sedation, α2 and α3 with anxiety, α6
with ataxia due to its cerebellar distribution, and α5 with learning
and memory, due to its hippocampal distribution.
Agonists bind to the usual cys-loop ligand binding region at the
α/β interface. By opening chloride channels, tonically active nerve
cells will generate fewer and less frequent APs. Hypervigilant
states and ultimately seizures result from GABAA antagonists such
as bicuculline (from Dicentra cucullaria or Dutchman’s breeches)
and securinine (叶秋 from Flueggea suffruticosa), long known to
Chinese and Native American herbalists. The action of GABA can
also be prevented by channel blockade, such as by picrotoxin (a
convulsant but also an herbal stimulant that was once added to
beer).
Three allosteric sites exist on the GABAA receptor. Benzodi-
azepines bind to a site at the α/γ interface and can have posi-
(PAMs) increase the receptor’s affinity for GABA, increasing
the frequency of channel openings. PAMs are effective anticon-
vulsants, sedatives, and anxiolytics, depending on the molecular
identity of the α subunit in the receptor and the location of the
receptor in the brain. Conversely, negative allosteric modulators
(such as thujone, the convulsant classically present in absinthe)
decrease the GABA affinity and the opening frequency; these
drugs are convulsants. Neutral (or silent) allosteric modulators
(such as flumazenil) have no effect on GABA affinity and are
used to reverse benzodiazepine overdoses by competitively bind-
ing to the allosteric site.
Barbiturates bind at a second allosteric site that is within the
channel and prolongs the channel open time, acting in synergy
with benzodiazepines. Other sedative-hypnotic agents, such as
propofol and chloral hydrate, also work at this site. Hence coin-
cident use (or abuse) of benzodiazepines with sedative-hypnotic
drugs is more than additive and requires caution. Death occurs
when the ventilatory drive is compromised.
A third allosteric site is also within the channel and binds vola-
tile anesthetics, ethanol, and neurosteroids to prolong open chan-
nel lifetime. The equivalent site on nAChRs blocks its channel.
Fortuitously, volatile anesthetics will thus depress the function
of the brain both by prolonging inhibitory receptors and blocking
excitatory receptors. 
Serotonin (5-HT) and Emesis
One of the seven serotonin receptors is a cys-loop ionotropic
receptor (5-hydroxytryptamine 3 [5-HT3]) with 5 types of sub-
units (5-HT3A through 5-HT3E). 5-HT3 receptors are distrib-
uted presynaptically and postsynaptically throughout the brain
and peripheral nervous system, particularly in the gut. Specific

antagonists effectively reduce the nausea caused by chemother-
apy and radiotherapy.  Effective
Cys-Loop Receptors at Targets in Parasites
Invertebrate nervous systems also use cys-loop receptors,
but their allosteric binding sites are distinctly different from
those found in vertebrate receptors. Ivermectin is a drug
that exploits this difference. Ivermectin is a potent PAM for
an inhibitory cys-loop receptor that is only found in insects
and worms, and so it has been used to kill the microfilariae
that lead to onchocerciasis (river blindness) and filariasis
(elephantiasis). 
G PROTEIN–COUPLED TRANSMITTERS
Signals from metabotropic receptors interact on multiple levels
and in many directions. To take one example, neurotransmitter
release is modulated by phosphorylation of proteins involved in
exocytosis, which is modulated by heterotrimeric G proteins
whose signals are modulated by the quantity of transmitter
released and whose second messengers modulate the kinases,
which phosphorylate the proteins of exocytosis. Almost all
aspects of brain function involve equally complex networks of
stimulatory and inhibitory controls, to perform the innumerable
highly specific and specialized tasks of which the nervous sys-
tem is capable. This section will present the important steps of a
framework of control mechanisms and how they are used for the
neurotransmitters already discussed, mindful that each and every
function of the nervous system will have its own combination of
control pathways, using its own selection of types, subtypes, and
isoforms of transmitter, receptor, controller, and effector pro-
teins and lipids.  GDP. GEFs are generated indirectly via second messengers such
Long-Term Potentiation by Phosphorylation
Repeated use of a synapse can lead to an increase in the quan-
tity of transmitter released (long-term potentiation [LTP])—one
of the mechanisms that are the basis of learning and memory.
Alternatively, the consequence of multiple stimuli may well be
a decrease in transmitter release (long-term depression [LTD]),
which is also a part of learning and memory. In both cases, pro-
tein phosphorylation is of crucial importance in determining syn-
aptic strength.
Many vesicle-release proteins are phosphorylated by phospho-
kinase A (PKA), the enzyme that is stimulated by cAMP, the
product of the adenylyl cyclase. This cyclase is stimulated by
Gs proteins and inhibited by Gi proteins. When PKA is active,
SNARE complexes are quicker to form, increasing the pool syn-
aptic vesicles at the active zone and strengthening the synaptic
response to a stimulus.
LTD results, in part, from the opening of potassium channels
that stabilize the membrane potential at a more negative poten-
tial. PKA increases the potassium permeability in two ways: by
increasing inward rectifier channel activity in nerve terminals and
by decreasing the duration of a burst of APs via ATP-sensitive
potassium channels.
Phosphorylation also works at one step removed: the RAS
superfamily of small G proteins (composed of the Ras, Rab,
Rho, Ran, and Arf families, Fig. 4.9) are activated when their
GDP (guanosine-5ʹ-diphosphate) is displaced by a GTP
(guanosine-5ʹ-triphosphate). This triphosphate alters the RAS
conformation so that it can bind to downstream targets, acti-
vating their (often kinase) activities. RAS family members are
small, often soluble proteins that have inherent GTPase activ-
ity, so they themselves hydrolyze their GTP to GDP and return
to the inactive state. The lifetime of GTP-RAS is abbreviated if
the GTPase is sped up, which can be done by GAPs (GTPase
activating proteins).
Chemical Signaling in the Nervous System 65
Active &
Rho Rho
GTP GTP
GAP
GDP
Effector
GEF
GTP GDI
Pi
Rho Rho
GDP GDP
Inactive &
GDI
Sequestered
Fig. 4.9  The small G protein in action. The Rho-type small G protein is inactive
when bound to guanosine diphosphate (GDP) and is stabilized in this inactive
state by guanosine nucleotide dissociation inhibitors (GDIs). The small G pro-
teins are activated when guanosine exchange factors (GEFs) displace the GDP,
allowing guanosine triphosphate (GTP) to bind and for the G protein to then
insert into the plasma membrane and interact with effector proteins. GTPase
activating proteins (GAPs) accelerate the rate of GTP breakdown, returning the
G protein to its inactive state. Pi, inorganic phosphate.
A signal (provided by proteins called GEFs, guanine nucleotide
exchange factors) is required to remove the tightly bound GDP,
freeing the RAS protein to substitute a different nucleoside. The
replacement is generally GTP, which is far more abundant that
as cAMP and calcium and directly by the growth factor receptor–
coupled tyrosine kinases. GDIs (guanosine nucleotide dissociation
inhibitors) prevent GEFs from displacing the GDP from Rho-type
kinases; hence they stabilize the inactive form of its small GTPase.
Small G proteins contribute to LTP in several ways. One exam-
ple is Epac, the GEF for the Rap G protein, which is directly
activated by cAMP and in turn increases the amplitude of synap-
tic currents. A second example is RAB3A, a small G protein that
can bind to the membrane of synaptic vesicles. Its GEF is GRAB
(guanine exchange factor for RAB3A). 
G Proteins Have Receptors as Exchange Factors
The large G proteins are heterotrimeric complexes (composed
of α, β, and γ subunits, Fig. 4.10) that build upon the function-
ality of small G proteins by having the ability to couple to G
protein–coupled receptors (GPCRs) (Table 4.3). The Gα subunit
is homologous to a small GTPase with its membrane anchor, a
GDI-like domain and a domain that allows the GPCRs to act as
GEFs. The Gβγ dimer is also membrane bound and always acts
as a pair.
Once again, the heterotrimer is inactive when GDP is bound
to Gα, the subunit with GTPase activity. When an agonist binds
to and activates a GPCR, it can act as a GEF, displacing the GDP
so that a GTP takes its place, causing the complex to dissoci-
ate into biologically active Gα and Gβγ subunits. Gα eventually
hydrolyzes the GTP to GDP, causing the heterotrimeric complex
to reassemble into an inactive trimer.
Three families of α subunits are activated by GPCRs: Gs (two
members), Gi/o (five) and Gq (five). Historically, the symbols
were differentiated according to their actions: “s” because it
stimulates adenylyl cyclase, increasing cAMP; “i/o” because it
inhibits adenylyl cyclase or has some other ability to directly acti-
vate certain potassium channels; and “q” for its queer or noncy-
clase-related ability to activate phospholipase C and initiate the
inositol and 2,3-diacylglycerol signals.
66 Essential Concepts

Agonist Group I metabotropic glutamate receptors increase cell cal-

binding cium by IP3 stimulation of calcium release from intracellular
stores, hence activating pathways and channels much like the
N-methyl-D-aspartate (NMDA) receptors.
R
Group II metabotropic glutamate receptors inhibit adenylyl
cyclase, reducing the cell cAMP. Reduced cAMP means less pro-
α tein kinase C, fewer cyclic nucleotide ion channels (i.e., CNG
β γ
and HCN) and less Epac, the GEF. Each of these actions can
have a profound impact on nerve function.
A
Group II/III metabotropic glutamate receptors have been
found to inhibit each class of calcium channels at one point
G protein coupling or another in the nervous system. In particular, inhibiting the
and nucleotide exchange P/Q type calcium channels on presynaptic membranes limits
the influx of calcium when that membrane is depolarized, thus
R*
reducing transmitter release from the nerve terminal. 

α
β γ
GTPGDP
B operate primarily by Gi and Gq). The prominent effects of antag-
Activated G protein subunits
regulate effector proteins
α AC β γ
ATP
K
+ ACTIVITY
cAMP
C Three biogenic amines are derived from phenylalanine: DA,
Subunits ready to recombine
α β γ
D Pi
Fig. 4.10  The trimeric G protein in action. At rest, the inactive Gαβγ protein is a
heterotrimer and binds a guanosine diphosphate (GDP) (A). A transmitter activates
the receptor that binds the G protein, inducing conformational changes in both mol-
ecules that increase how tightly they bind each other (B). Furthermore, the bind-
ing triggers the release of the GDP and its replacement by guanosine triphosphate
(GTP). (C) The Gα subunit dissociates to diffuse to effectors, such as the adenylyl
cyclase shown here. The Gβγ diffuses to its targets, such as this G protein–coupled
inwardly rectifying potassium (GIRK) channel. (D) With the hydrolysis of the bound
GTP, activity ceases and the heterotrimer reassociates. ATP, adenosine triphosphate;
cAMP, cyclic adenosine monophosphate; Pi, inorganic phosphate.
The Gβγ subunits have biologic activities as well. Once dissoci-
ated from Gα, the Gβγ subunit activates many inwardly rectify-
ing potassium channels as well as a variety of adenylyl cyclase,
kinase, and phospholipase molecules.
All G protein receptors have seven transmembrane segments
(7TM). Four of the segments bind the agonist—the orthosteric
modifier of the protein). Transmembrane segments are also used
to propagate the agonist binding to the ultimate opening of the G
protein binding site.  adrenaline, derive from two lines of 19th-century research.
Glutamate and the G Proteins
Metabotropic glutamate receptors operate by three distinct
mechanisms: by activation of GIRKs (the G protein–coupled
inwardly rectifying potassium channel, via Gβγ), by increasing
cell calcium (via Gq), and by inhibition of adenylyl cyclase
(via Gi).
Muscarinic Receptors
The metabotropic receptors for acetylcholine are all activated
by muscarine, the poison of the Amanita muscaria mushroom,
hence the name muscarinic. Five receptor types (M1 to M5)
onists such as atropine and scopolamine are primarily due to
alterations in the function of the autonomic nervous system: dry
mouth, slow heart, dry skin, and vasodilation. Central effects are
largely due to M1 receptors and are most commonly apparent as
the untoward side effects of psychotropic medicines: confusion,
disorientation, and hallucinations. 
THE BIOGENIC AMINES MODULATE SYNAPTIC
norepinephrine (NE), and epinephrine (Epi) (Fig. 4.11). Two
others are derived from l-tryptophan (serotonin or 5-HT) or
histidine (histamine), respectively. Of these precursors, only
phenylalanine has potential toxicity: Approximately 1% of
Caucasians have a defective phenylalanine hydroxylase gene.
Homozygotes for the defect cannot transform phenylalanine
to tyrosine (Fig. 4.11), leading to high serum, urine and cere-
brospinal levels of phenylalanine (PKU or phenylketonuria).
The cerebrospinal fluid of individuals with unmanaged PKU
will contain millimolar phenylalanine, at which concentration
amyloid fibrils spontaneously form. These fibrils are immu-
nogenic and hence cytotoxic, leading to severe neurological
deficits.
Acetylcholine and amino acids are not included in the
group we call biogenic amines, even though choline contains
a quaternary amine and amino acids are, by definition, amine-
containing acids. The term biogenic amine has long served a
valuable purpose because the transmitters in this group have
similar synaptic machinery and receptor behavior, and they
serve our purpose well by acting as examples of transmit-
ter uptake and disposal mechanisms and the consequences of
having presynaptic and postsynaptic receptors activated by
a single transmitter. Functionally, they do not directly stim-
ulate or inhibit the postsynaptic cell, as do glutamate and
GABA, but instead they increase or decrease, that is, modu-
late, activity at the synapse via G protein–coupled effectors
(Fig. 4.12).
The catechol terms used in this chapter, epinephrine and
Adrenaline was the pharmacologist’s adrenal gland extract that
had the functional ability to increase heart rate and blood pres-
sure. Hence functional structures have names based on the word
adrenaline. Epinephrine was a chemist’s concept that required
many years of refinement to become the molecular structure
that we now know by name. So we can buy epinephrine, but we
talk about adrenergic synapses.
 Chemical Signaling in the Nervous System 67

Phenylalanine Fig. 4.11  The catecholamine family of chemical structures. Phenylalanine (F)
is an essential amino acid that is hydroxylated to form tyrosine (Y) by phenylala-
nine hydroxylase. The catechol ring (purple) is completed by tyrosine hydroxylase
O adding the second hydroxyl group, making l-DOPA. Tetrahydrobiopterin (BH4)
is the cofactor for both reactions. Removal of the carboxyl group by DOPA decar-
boxylase and the cofactor pyridoxal phosphate yields dopamine. Dopamine is
converted to norepinephrine in synaptic vesicles by dopamine β-monooxygenase
(or dopamine β-hydroxylase). Norepinephrine is methylated in the adrenal glands
to form epinephrine by phenylethanolamine N-methyltransferase with S-adenosyl
methionine as the methyl donor. Highlighted are the sites where the chemical
H N OH structures have been modified. l-DOPA, l-3,4-dihydroxyphenylalanine.

H
Tyrosine

HO O
Catechol Synthesis and Storage
Chemically, the brain is a protected environment where only
lipid-soluble molecules (general anesthetics, ethanol, heroin, nic-
otine, diazepam, etc.) freely enter. Molecules that cannot freely
H N OH diffuse through the membrane’s lipid bilayer are restricted by
the very special structure and function of the blood vessels that
OH H
supply nutrients to the brain, the blood-brain barrier (see Chap-
ters 2, 6, and 8). Hence water-soluble molecules must enter by
special carrier transport systems. For instance, glucose (via the
L-DOPA
glucose transporter GLUT1, Table 4.2), and ketone bodies (via
monocarboxylic acid transporter) can enter and provide energy
to the brain, but fatty acids cannot. Phenylalanine, tyrosine, and
HO O l-DOPA (l-3,4-dihydroxyphenylalanine) can enter (via the large
neutral amino acid transporter) but the small neurotransmit-
ters (ACh, the catecholamines, glutamate, serotonin, and hista-
mine) cannot. Unlike the cholinergic system, much of a synaptic
vesicle’s catecholamine is recycled from the synaptic cleft and
H N OH not from newly synthesized molecules. However, the mecha-
OH
nisms involved in the synthesis and storage of catecholamines
H are important clinically because they provide sites to intervene
therapeutically.
Dopamine
Synthesis of l-DOPA (via tyrosine hydroxylase, Fig. 4.11) is the
rate-limiting step in catecholamine production, allosterically inhib-
ited by DA and the other catecholamines. The potent enzyme
HO inhibitor α-methyltyrosine will therapeutically deplete DA stores
H and thus protect against the catecholamine storms released by
pheochromocytomas. Additionally, oxygen is consumed in DOPA
N
synthesis, and hydrogen peroxide can be produced if the reaction
H is partially uncoupled, generating the risk of oxidative stress. This
can damage nearby cells and contribute to Parkinson disease (PD).
OH
In PD the production of DA is severely compromised and
needs to be augmented; l-DOPA therapy is central to PD treat-
ment because the rate limiting step is bypassed. However,
Norepinephrine OH
l-DOPA works throughout the body, and so there are unwanted
side effects due to excess peripheral DA. Fortunately, these side
HO effects can be minimized by using a tyrosine hydroxylase inhibi-
H tor that cannot cross the blood-brain barrier such as carbidopa
and levodopa.
N Vesicular storage of catecholamine neurotransmitters shares the
mechanism that is present in cholinergic vesicles. An homologous
H
OH protein (VMAT2, Table 4.2) of the same transporter family uses
the hydrogen ion gradient to concentrate the positively charged
catechols within the vesicle. Glutamate is frequently the coun-
Epinephrine OH terion because VGLUT is commonly present in the membrane
of the catecholamines vesicle (Fig. 4.2). Release of these dual-
neurotransmitter synaptic vesicles results in dual-receptor effects.
HO Reserpine, a specific VMAT2 inhibitor, markedly reduces the
H
catecholamine content in these vesicles, inhibiting the action
N
of postganglionic sympathetic nerves and generally depleting
catecholamines throughout the brain. A similar blocker (tetra-
benazine) is used therapeutically to reduce the hyperkinetic
movements of Huntington chorea, Tourette and other tic syn-
dromes, tardive dyskinesia, and spontaneous hemiballismus.
68 Essential Concepts

Norepinephrine release Norepinephrine plus peptide release

Tyr

L-DOPA Large
dense-cored
synaptic vesicle Axon
Dopamine (DA)
NE
+
DA NE Small Peptides
clear
synaptic vesicle

α2-Adrenergic
receptor
Nerve
terminal

NET NE
NE
NE +
Peptides Synaptic
cleft
MAO
Plasma
membrane
EMT β-Adrenergic Class B
receptor G protein–coupled
receptor Postsynaptic
neuron

Fig. 4.12  Modulation of synaptic function: the noradrenergic (NE) synapse. NE synapses release both small
clear synaptic vesicles (SVs) and much larger dense-cored vesicles (DCVs). The SVs store NE, whereas the
larger vesicles stain densely because they contain granin proteins and peptide neurotransmitters as well. Clear
synaptic vesicles (left): Catechol precursors are synthesized locally, and NE itself is synthesized within the
SV. However, most vesicular NE is recycled from used transmitter that is recovered from synaptic cleft by
special transporters (NET) and reloaded locally. Any excess catecholamine is degraded by monoamine oxi-
dase (MAO) present on mitochondria in the nerve terminal and on the target cell as well. NE also exits the
cleft into the target cell (via EMT). Release of the small vesicles is regulated by calcium, and a single action
potential can elevate the calcium concentration in the nerve terminal sufficiently to trigger the release of
SVs. Release can be full and complete, with vesicle membrane retrieved by clathrin, reforming empty vesicles
that are then refilled. Alternatively, vesicle fusion proceeds only as far as the formation of the narrow pore,
transiently releasing only a fraction of their contents (kiss-and-run vesicle fusion). NE acts at target cells
(β receptor) and back on its own nerve terminal (α2 receptor) via class A metabotropic G protein–coupled
receptors. Dense core vesicles (right): DCVs exocytose from the axon or (more commonly) from the nerve
terminal only after an intense period of stimulation, after a train of action potentials. The peptides of DCVs
cannot be produced locally, and once the vesicle’s contents are lost by full vesicle exocytosis, the recovered
DCV membrane is returned to the neuron’s soma for lysosomal processing. However, kiss-and-run events
only release NE because the fusion pore is too narrow for the peptides to leave; in this case, the DCVs can
refill and remain in the nerve terminal for later use. When released, the neuropeptides bind to class B metabo-
tropic G-coupled receptors that have an expansive ligand binding site—large enough to recognize a molecule
the size of a peptide. This signal is then terminated by peptide hydrolysis via synaptic cleft endopeptidases.

Norepinephrine (NE) synapses differ from DA synapses
because they contain the enzyme dopamine β-hydroxylase
(DBH, Fig. 4.12). Unlike any other enzymes involved in neu-
rotransmitter synthesis, DBH is membrane bound within the
synaptic vesicle, and DA must be transported into the vesicle to
be converted to NE. In cells that secrete Epi, NE must exit the
vesicle, be methylated by phenylethanolamine-N-methyl trans-
ferase, and the resulting Epi returned to the vesicle for storage
and release.  vagal afferent neurons, which leads to activation of brainstem
Serotonin Synthesis and Functions
Serotonin (5-HT, 5-hydroxy-tryptamine) is derived from the
essential amino acid tryptophan (Fig. 4.13). Tryptophan is gener-
ally safe as a dietary supplement, but toxic material generated dur-
ing the fermentation and manufacturing processes has not always
been removed during the purification steps. In 1989, one sup-
plier’s process did not remove 1,1’-ethylidenebis[tryptophan],
which lead to a widespread outbreak of an eosinophilia-myalgia
syndrome, with associated muscle pain and weakness.
Within the CNS, serotonin is found in the dorsal raphe, a
nucleus that projects throughout much of the brain, regulating
mood in many important ways. However, most of the body’s
serotonin is in the gastrointestinal tract, stored in entero-
chromaffin cells and normally used to coordinate gut func-
tion. Severe nausea and vomiting accompany many forms of
radiation exposure and chemotherapy due to the sustained
release of serotonin from enterochromaffin cells, activating
emesis centers. Antiemesis drugs, such as palonosetron, bind
specifically to the extracellular domain of 5-HT3 receptors,
blocking their function and causing them to be removed from
the surface by internalization. A delayed wave of nausea and
vomiting is caused by signals from the area postrema that
release the neuropeptide substance P from vagal efferent neu-
rons. This peptide binds to NK1 receptors that amplify the
serotonin effects, so the specific NK1 inhibitor aprepitant is
generally paired with a 5-HT3 antagonist in the treatment of
nausea and vomiting.

L-Tryptophan
O fusion pores as well as full fusion with the plasma membrane.
OH
HN NH2
HO
Serotonin
HN NH2
HO
5-Hydroxyindoleacetic acid
OH
O NE is transformed into normetanephrine, Epi to metaneph-
HN
Fig. 4.13  The serotonin family of chemical structures. Serotonin does not cross
the blood-brain barrier; it must be synthesized and later enzymatically degraded
by the neuron. Synthesis begins by adding a hydroxyl group to the indole ring
of the essential amino acid tryptophan (W) via the rate limiting enzyme trypto-
phan hydroxylase, making 5-hydroxy-l-tryptophan. Second, the carboxyl group is
removed via AADC (the aromatic amino acid decarboxylase), resulting in a mol-
ecule of serotonin (5-HT, 5-hydroxytryptamine). Catabolism is by monoamine
oxidase to 5-hydroxyindoleacetic acid (5-HIAA) and hydrogen peroxide, a reac-
tive oxygen species. The altered side groups are highlighted.
Toxic serotonin effects are manifest in the gut, vasculature, and
brain. Ergotamine, a serotonin agonist, contaminates seed grain
growing the Clavicepts spp. fungi. Eating tainted food leads to
vomiting, spastic contractures, delirium, and gangrene. In the past,
entire regions were affected as their grain stores molded, with
outbreaks of ergotism being marked by dramatic mass hysterias,
residual pathologies, and witch persecutions. The hysterias were
similar to the hallucinations caused by lysergic acid diethylam-
ide (LSD), psilocybin, and mescaline, all serotonin agonists that
act at the 5-HT2A/2C receptors. As with other food poisoning (or
even modern chemotherapy), excess gut 5-HT leads to diarrhea
and vomiting via 5-HT3 receptors. The ergot-related gangrene was
due to serotonin’s potent and long-lasting vasoconstrictor effects
(acting via 5-HT1B/1D receptors). Indeed, even today ergotamine is
used to constrict cerebral vessels during migraine attacks.  The DAT, NET, and SERT transporters are not very specific
Catechol Release and Reuptake
As with the NMJ, the release of clear and dense core vesi-
cles that contain catecholamines use SNARE complexes and
synaptotagmins to realize calcium-dependent exocytosis at
active zones on the nerve terminal. Also like the NMJ, clear
Chemical Signaling in the Nervous System 69
and dense core vesicles both exhibit kiss-and-run transient
However, unlike the NMJ, most of the released catecholamine
is taken back into the presynaptic terminal, to be returned
to vesicle storage and not destroyed enzymatically in the syn-
aptic cleft. A low-affinity, high-capacity monoamine trans-
porter (plasma membrane monoamine transporter [PMAT])
removes much of the transmitter from the synaptic cleft. In
addition, there are high-affinity specific transporters for each
of the catecholamines, expressed by the neuron according its
own particular transmitter: DAT for the dopamine transmit-
ter, NET for norepinephrine transmitter, and SERT for sero-
tonin transmitter (Table 4.2). Any catecholamine that is not
recycled into the nerve terminal is oxidized by postsynaptic
MAO-B, the B-type monoamine oxidase.
A number of safeguards are built into this simple but highly
effective system because any unconfined catecholamine is
likely to have untoward effects. The catecholamine transport-
ers operate with a high throughput capacity, but they cannot
concentrate effectively. In fact, they slip into a reverse mode
if the terminal catecholamine concentration grows too large,
spilling catecholamine back into the synaptic cleft. Hence
the nerve terminals have MAO-B linked to the outer mem-
brane of mitochondria to oxidize any excess catecholamine
in the cytoplasm (Fig. 4.12). Similarly, excess catecholamine
that escapes the synaptic cleft is transported into neighbor-
ing glial cells, out of the cerebrospinal fluid by endothelial
cells of the vasculature or—in regions of the brain with lit-
tle MAO, such as the prefrontal cortex—by the postsynap-
tic cell. Glia, endothelia, and the postsynaptic membrane
use a different transporter, the extraneuronal monoamine
transporter (EMT). Once internalized by the glia, endothe-
lia, or a postsynaptic cell, the catecholamine is inactivated by
membrane-bound catechol-O-methyl transferase (COMT).
rine, DA to 3-methoxytyramine and significantly, l-DOPA to
3-O-methyldopa. This last reaction can markedly reduce the
effectiveness of l-DOPA given for treatment of Parkinson dis-
ease; hence triple therapy for Parkinson disease has become
l-DOPA plus a COMT inhibitor and a tyrosine hydroxylase
inhibitor that cannot cross the blood-brain barrier. 
Psychotropic Drugs and the Uptake and Delivery of
Catecholamines
Drugs that act at catecholamine neurons in the brain generally
alter mood and behavior to a marked degree. Two of the stron-
gest drugs block catecholamine transport. Cocaine blocks cat-
echolamine uptake by DAT and, to a lesser extent by NET and
SERT, causing the synaptic cleft to be flooded by catecholamine
for an extended period after each presynaptic AP. This leads
to euphoria and a hyperalert state of mind. Peripheral mani-
festations of this prolonged catecholamine excess is evident in
the ischemic loss of nasal tissue at the site where the drug is
insufflated, resulting in α2-adrenoreceptor–mediated constric-
tion of nutrient blood vessels. Other peripheral manifestations
of cocaine’s vasoconstrictor effects are cardiac arrhythmias,
abruptio placenta, and cardiac ischemia. CNS effects can esca-
late through the ranks of anxiety, centrally mediated impo-
tence, depression, episodes of uncontrolled rage, and ultimately
psychosis.
with regards the structure of their cargo, as they will transport a
variety of molecules, such as the amphetamines, into the nerve
terminal. Once in the neuron, amphetamines block the synaptic
vesicle’s VMAT2 transporter and eliminate the proton gradient,
preventing storage of catecholamine in synaptic vesicles. Because
the catecholamine cannot be stored, it leaves the terminal by
70 Essential Concepts
reverse transport through these DAT, NET, and SERT transport-
ers, to flood their synaptic cleft. The results are similar to those
of cocaine, but far more toxic to DA and serotonergic neurons in
the striatum, hippocampus, and prefrontal cortex.
A second example that is rapidly fatal to DA neurons is
MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine), a syn-
thetic opioid. Once MPTP arrives at a synaptic cleft, MAO-B
converts the drug into MPP+ (1-methyl-4-phenylpyridinium),
a specific mitochondrial toxin that is transported by DAT into
the presynaptic nerve terminal, rapidly leading to cell death.
Cell loss is substantial in the substantia nigra pars compacta and
other regions of the brain where DAT is present. MPTP was
first synthesized in 1947 and soon shown to cause severe par-
kinsonian signs and to be fatal in human subjects. MPTP toxic
effects are seen with use of the street drug desmethylprodine
or MPPP (1-methyl-4-phenyl-4-propionoxypiperidine), a psy-
choactive analog of meperidine, a Schedule I opioid. Although
MPPP itself is not toxic, MPTP is a byproduct of its synthesis,
and so casual users of MPPP developed profound parkinsonian
signs and symptoms.
Therapeutic drugs that inhibit catecholamine uptake are
anxiolytic mood elevators with some antipsychotic effect. The
drugs are generally SERT inhibitors (hence the name SSRI, or
selective serotonin reuptake inhibitors), although they also
inhibit NET and DAT to varying degrees, depending on their
molecular structure. Unlike cocaine, whose action is immedi-
ate, the clinical utility of SSRIs is apparent days to weeks after
therapy has begun. During this time, the depleted presynaptic
stores of transmitter have returned toward normal as synthesis
is augmented and postsynaptic (and presynaptic) receptors have
also adjusted to the increased synaptic cleft catecholamine lev-
els. Given the multiplicity of diseases involved and the innate
variability of the subjects treated, not one of these changes
seems to completely explain the multiple actions of SSRIs.  function of viscera, growth of the organism, and response to
G Protein Receptors for the Biogenic Amines
With one exception, the ionotropic 5-HT3, all receptors for the
biogenic amines couple to G proteins or to β-arrestin, with speci-
ficity largely determined by the cellular and anatomic location
of the receptor and the availability of the downstream effectors.
Receptors are generally postsynaptic, but α2 and certain 5-HT1
receptors are presynaptic or present in the cell body where recur-
rent collaterals regulate AP generation.
G proteins couple to the receptors for biogenic amines in a
characteristic manner: Dopamine receptors couple to Gs (D1 and
D5) and Gi/o (D2, D3, and D4). D2 receptors are important phar-
macologically: Antipsychotic drugs are D2 and 5-HT2A antago-
nists. D2 agonists can treat dyskinesia. Norepinephrine receptors
couple to Gq (α1), Gi/o (α2 and β3), and Gs (β1, β2, and β3).
Serotonin receptors couple to Gi/o (5-HT1), Gq (5-HT2), and Gs
(5HT4, 5-HT5, 5-HT6, and 5-HT7). 5-HT1D agonists are used to
treat migraines, and 5-HT3 antagonists treat emesis.
Depending on the agonist used, the coupling of some catechol-
amine receptors can be shifted from G proteins to β-arrestin,
activating mitogen-activated protein kinase, increasing the release
of cell stores of calcium, and prompting the internalization of the
cell surface receptor and consequent loss of the cell’s response
to that agonist.  mitters (molecules acting locally). Oxytocin and vasopressin are
PURINE NEUROTRANSMISSION: SENSATION AND
COFFEE TOO
Purines (ATP, adenosine diphosphate [ADP], and AMP) act
through both ionotropic (P2X) and metabotropic receptors (P1
for adenosine and P2Y for ATP, ADP, and AMP). Purines need no
transmitter-specific synthetic pathways; they are necessary for
the existence of the cell, so ATP will be released during ischemic
periods through the large pannexin and connexin channels, and
at cell death. ATP is also released as a counterion to cation neu-
rotransmitters. Hypoxia and acid hypercapnia releases ATP from
glomus cells at the carotid body, stimulating afferent nerve fibers
to the breathing control centers of the brainstem. Finally, type
II taste bud cells for sweet, bitter, and umami sensations release
ATP through a voltage-gated ion channel (calcium homeostasis
modulator 1 [CALHM1]) to signal the afferent gustatory nerve
pathways to the brain.
ATP is short lived outside of the cell as it is enzymatically
degraded to ADP, AMP, and adenosine by membrane-bound
ectoenzymes and returned to the cell by equilibrative nucleoside
transporters (ENT).
P2X receptors are homotrimers and heterotrimers that form a
central pore accessible by vestibules within the large extracellular
domain. Being a large (7 Å), low field strength pore, the open
channel is permeable to monovalent and divalent cations. When
located presynaptically, this calcium influx greatly increases
transmitter release. Postsynaptic P2X receptors in afferent nerve
fibers are activated as part of taste, hypoxia, hypercapnia, and
pain sensations.
Purine receptors are coupled to Gi/o (A1 and A3) and Gs (A2A
and A2b) proteins. A1 receptors are widespread in the brain and
are thought to mediate the stimulatory effects of caffeine, an
antagonist at this receptor. A2 receptors reciprocally inhibit the
actions of DA such that A2 antagonists reduce some symptoms
of Parkinson disease. 
PEPTIDES MODULATE BRAIN FUNCTION
Peptides are active in the brain as neuropeptides, growth fac-
tors, and cytokines. Much of their function is beyond the scope
of this chapter because they modify integrative functions of
the nervous system—states of sleep/awake, hunger/satiety, and
compassion/stress, for instance. In addition, they modulate the
inflammation.
More than 40 neuropeptides have been identified in brain.
Neuropeptides are synthesized in the form of large precursor
peptides that are processed in the endoplasmic reticulum and
Golgi apparatus to be packaged as DCVs. Alternative RNA
splicing and differential cleavage of the peptides yield a wide
variety of bioactive neuropeptides. Maturation of neuropep-
tides includes methylation, sulfation, or glycosylation steps and
occurs throughout the period of time between transcription of
the gene to the point where the DCVs are delivered to the
neuron’s terminal.
Neuropeptides act both as neurotransmitters to modulate syn-
aptic function and as hormones, to act at a distance. The peptides
that act at the Y receptor are the most common neurotransmit-
ters, with neuropeptide Y (NPY) acting centrally to stimulate
appetite and peripherally to augment the effect of its co-released
NE. Other complex systems of transmitters and their receptors
modulate reward (opioid receptors), wakefulness and appetite
(orexins), and pain (tachykinins).
Hypothalamic peptides include a set of neurohormones that
are released for uptake by vessels in the median eminence to con-
trol the adenohypophysis, demonstrating the overlap between
hormones (signal molecules released into the blood) and trans-
neurotransmitters in many sites in the brain, but magnocellular
neurons of the hypothalamus project to the neurohypophysis
(the posterior pituitary gland), where these peptides are released
and then taken up and stored for later release as neurohormones. 
RETROGRADE TRANSMISSION
Most neurotransmitters work in the forward direction, with their
chief effect on the postsynaptic receptors, although in special
cases they also exert negative feedback on their own release

via presynaptic receptors. Cannabinoids, however, always act
backward, being produced by the postsynaptic cell to act at the
presynaptic nerve terminal. This is called retrograde transmis-
sion. A strong calcium signal is required to activate the synthetic
enzymes for the endocannabinoids in the cells where this system
is present. Being lipid soluble, the cannabinoids diffuse through
the postsynaptic membrane, back into the presynaptic terminal
and other surrounding cells to activate Gi-coupled cannabinoid
receptors. These are the receptors that are activated by the active
components of marijuana.
Calcium signals can also activate the nerve form of nitric oxide
synthase, resulting in nitric oxide (NO) production. Again, being
lipid soluble, NO diffuses back into the presynaptic nerve ter-
minal, where it activates soluble guanylyl cyclase and generates
cyclic guanosine monophosphate. One consequence of elevated
cGMP is that synaptic vesicle trafficking from the Golgi appa-
ratus to the nerve terminal is inhibited, which depletes the pool
of vesicles at the synapse and thus decreases neurotransmitter
release.  This chapter has provided signposts in a vast, imperfectly
CODA
Throughout this chapter, a framework of the nervous system has
been presented that has no beginning and no end, with a dozen
classes of transmitters, each having multiple receptors. Much of
Chemical Signaling in the Nervous System 71
the chemical signaling described involves modulation of neuronal
function that uses many of the 800-plus G protein receptors as
well as numerous other controllers, with various molecules com-
partmentalized by a dozen different solute carrier transporters.
Moreover, the system is dynamic, with change occurring every-
where and all the time.
In reality, the brain is more complicated than this.
Receptors are not neatly divided by their transmitter. Hetero-
meric receptors form between Gq-coupled serotonin 5-HT2A
receptor and the Gi/o-coupled metabotropic glutamate 2 recep-
tor such that glutamate transmission activates Gq-coupled
effectors.
Psychotropic drugs rarely work at a single transmitter family
of receptors but act broadly in ways that vary from one drug to
the next.
Age matters, with the same system being excitatory—and then
inhibitory—in neonatal, childhood, adolescent, young adult, and
aging brains.
understood system and has provided generalizations that have
exceptions. It may always be thus.
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The complete list is available online at www.studentconsult.com.
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