Autonomic Nervous System:

Introduction to Neurotransmitters and

Receptor Specificity
Knowledge objectives introduced by this lecture:
Identify the key conceptual similarities and differences
between autonomic cholinergic and adrenergic pathways
including receptor subtypes, neurotransmitters, transmitter
synthesis, storage, and release, and relative specificities of
drugs that stimulate or inhibit each branch or activity.
List the major systems or organs innervated by the
autonomic cholinergic and adrenergic systems.
Describe the organ system effects of cholinergic and
adrenergic stimulation or antagonism.
Relate the tissue expression profiles of cholinergic and
adrenergic receptors to their specific functions.
Body functions are controlled by two major systems:
(1)The endocrine system, which secretes hormones.
(2) The nervous system, and this is subdivided into:
Central nervous system(CNS),consisting of brain
and spinal cord.
Peripheral nervous system(PNS) consisting of:
sensory(afferent), take information to CNS.
motor (efferent) take informations (orders) to
tissues &organs.
The motor is subdivided into somatic and
autonomic.
The autonomic is subdivided into
sympathetic and parasympathetic.
The ANS is largely independent.ie its
functions are not under direct conscious
control, eg: contraction and relaxation of
vascular and visceral smooth muscles, all
exocrine and some endocrine secretions, heart
beats and energy metabolism.
 The ANS uses chemicals (neurotransmitters) for
transmission of information.
Transmission occurs between nerve cells and
between nerve cells and their effector cells.
This is achieved by release of chemicals
(neurotransmitters) from nerve cells into synaptic
cleft that diffuse and activate or inhibit post synaptic
cells by binding to specific receptors.
It is noteworthy that a single preganglionic neuron
synapeses with several postganglionic neurons, thus
spreading effect over a larger area.
The Sympathetic and Parasympathetic nerves
and their relation to each other
 Sympathetic Innervation
Preganglionic sympathetic nerves arise from cell bodies in the
thoracolumbar segment of the spinal cord.

They synapse in paravertebral sympathetic ganglia arranged in

two chains lying on both sides of spinal cord. These ganglia
contain cell bodies of postganglionic nerves.

However some preganglionic fibers do not synapse in

paravertebral ganglia, but pass through them to synapse in
collateral ganglia, eg coeliac, superior mesenteric, and inferior
mesenteric ganglia.
 Postganglionic nerves arise from these cell bodies
and innervate different organs of the body.
The only exception to this two-neuron arrangement
is the innervation of the adrenal medulla.
The catecholamine- secreting cells of adrenal
medulla are modified post ganglionic sympathetic
neurons, and the nerves supplying the gland are
equivalant to the preganglionic fibres
The adrenal medulla secretes a mixture of
epinephrine(70%) and norepinephrine(30%).
Parasympathetic innervation
Preganglionic parasympathetic nerves arise from cells in the
craniosacral segments of the spinal cord(III,VII,IX &X)
cranial and the III and IV sacral spinal roots.
Postganglionic nerves arise from ganglia located in or near
the organ innervated, thus they are very short.
The enteric nervous system consist of neurons whose cell
bodies lie in the intramural plexuses in the wall of the
intestine. It receives inputs from sympathetic and
parasympathetic systems, but can act on its own to control
the motor and secretory functions of the intestine.
 In some sites such as gut and bladder,the
sympathetic and parasympathetic produce opposite
effects.
In other sites only one division of autonomic nerves
is present. eg sweat glands and most blood vessels
have only sympathetic innervation, whereas the
ciliary muscle of eye has only parasympathetic
innervation, but there are adrenoceptors.
Bronchial smooth muscle has only parasympathetic
(conistrictor) innervation, but only 2-adrenergic
receptors that mediate relaxation.
 Resistance arteries have sympathetic conistrictor
innervation, but no parasympathetic innervation, instead
relaxation is mediated by release of nitric oxide from
endothelial cells that contain cholinergic receptors.
In the salivary gland and other organs the two systems
produce similar rather than opposing effects.
Thus the two systems are not always physiological
opponents.
Sympathetic activity increases in stressfight or flight
response, whereas parasympathetic activity predominates
during rest and digest.
Autonomic Innervation
Pharmacological division of cholinergic vs. adrenergic
neurotransmission
All preganglionic autonomic and postganglionic
parasympathetic neurons use acetylcholine(Ach) as
neurotransmitter. Ach is the neurotransmitter at
autonomic ganglia,smooth muscle tissue synapses,
cardiac muscle and motor nerve to NMJ.
Most postganglionic sympathetic fibers release
norepinephrine as the neurotransmitter which acts on -
or - adrenoceptors.They are called adrenergic fibers.
Thereare exceptions: Cholinergic transmission in
sympathetic system(all ganglia), adrenal medulla, sweat
glads use Ach (nicotinic or muscarinic). Dopaminergic
innervation in sympathetic system renal blood vessels.
Synapse sites most amenable to pharmacologic manipulation:

Na +
Precursors
(choline/tyrosine)

Synaptic
Precursor cleft
Neurotransmitter

Pre-synaptic Storage
nerve cell Release
Recognition
Ca 2+ by receptors
Metabolic
disposition Post-synaptic
nerve cell

Manipulation possible at pre-synaptic neuron: where neurotransmitter

is synthesized, stored and released upon cell activation; or at post-
synaptic neuron or effector cell: where neurotransmitter is detected
and its action is translated into cellular activities.
 Key Steps in Neurotransmission:

Synthesis & Storage Metabolism

Action Release Recognition

potential
(action)

Reuptake
Strategies for Pharmacological Intervention:
Block synthesis and storage: -usually rate-limiting steps; produce long-term effects
Block release: -rapid action and effective
Block reuptake: -increases synaptic neurotransmitter concentrations

- can be selective or non-selective

Interfere with metabolism:
- can be reversible or irreversible; blocking metabolism
-increases effective neurotransmitter concentration
 Definition of Agonist and Antagonist:
Agonist: (1) A natural ligand that activates a receptor.
(2) A drug that has properties similar to a natural ligand in activating the same
receptor.

Antagonist: (1) A receptor-specific blocker. (2) A molecule, such as a drug (e.g., enzyme
inhibitor) or a physiologic agent (e.g., hormone), that diminishes or prevents the action of
another molecule.

Mode of Action:
Direct-acting: Molecule that physically binds to the target for its effect.
Example: carbachol activates cholinergic receptors,atropine blocks them

Indirect-acting: Molecule that exerts effect on the target by interacting with

another non-target site.
Example: neostigmine blocks AchE, causing Ach accumulation.

Mode of action and agonism are different concepts. For example, a direct-
acting molecule can be either agonistic or antagonistic.
 Otto Loewi (Nobel Laureate, 1936)

Discovered that stimulation of the vagus (parasympathetic nerve) of a

frogs heart causes release of a substance that, when applied to a second
heart, could slow heart rate. He called this substance Vagus stoff,
demonstrating the chemical basis of neurotransmission.

He also found that atropine can prevent the inhibitory action, but not the
release, of Vagus stoff.

Exposure of Vagus stoff to frog heart homogenate inactivates it.

Physostigmine enhances the effect of vagus stimulation on the heart, and

prevents the destruction of Vagus stoff.
 Synthesis of acetylcholine:

Choline Acetylcholine O
CH3 CH3
Choline CH3 N + -CH2CH2O- CCH3
CH3 N + CH2CH2OH acetyltransferase
CH3
CH3 + +
O
CoASCCH3 CoA-SH
Acetyl-CoA CoA
(from mitochodria)
 Synthesis, storage and release of acetylcholine:

Na+
Choline
(10 M)

Choline
Ac-CoA ChAT Synaptic
cleft
Antiporter
Ach
Nerve Ach choline
impulse Ach+ acetic acid
Ach Ach
NN
Pre-synaptic Ca2+Ach AchE
cell
Recognition
Ca2+ by receptors
NM
CAT = choline acetyltransferase Post-synaptic
AchE
AchE = acetylcholinesterase cell
Degradation of acetylcholine:

H2O

O Choline Acetic acid

AchE
(CH3)3 N+CH2CH2O CCH3 (CH3)3 N+CH2CH2OH+ + CH3COOH
(-) OH
AchE
Glu202 Ser203
Tyr337 Glu334
His447

Steps involved in the action of acetylcholinesterase:

1. Binding of substrate (Ach)
2. Formation of a transient intermediate (involving -OH on Serine 203, etc.)
3. Loss of choline and formation of acetylated enzyme
4. Deacytylation of AchE (regeneration of enzyme)
Drug intervention in Cholinergic transmission

(Rate-limiting) Precursor transport Hemicholinium

: Stimulatory
Synthesis : Inhibitory
Cholinergic antagonists Solid: Agonistic
Dotted: Antagonistic
Atropine (anti-M)
d-tubocurarine Storage Vesamicol
(anti-NM)
Trimethophan
(anti-NN)
Release Botulinum toxin
Cholinergic agonists
(direct acting) AntiChE
Carbachol Ach Reversible (neostigmine)
Pilocarpine
Irreversible (organo-
Receptor Degradation phosphate)
+ action by AchE
 An example of indirect agonism:

Physostigmines effect on acetylcholine receptor is indirect.

This effect is mediated through the inhibition of cholinesterase,
which causes an increase in the local concentration of
acetylcholine. The net effect is agonistic on acetylcholine
receptor.
 Julius Axelrod (Nobel Laureate, 1970)
His discoveries concern the mechanisms which regulate the formation of
norepinephrine in the cells and the mechanisms which are involved in the
inactivation of this important neurotransmitter

Synthesis of Catecholamines Tyrosine hydroxylase

1
3 CH2 HO CH2
TH
HC COOH HC COOH Dopa decarboxylase
Phenylethanolamine-
HO HO
(L-amino acid
NH2 NH2
N-methyl transferase Tyrosine DOPA decarboxylase)
OH DD (L-AAD)
OH
HO
CH
HO CH2
HO CH PNMT
CH2 DBH CH2
CH2 HO
HO
HO
NH2 NH2
NHCH3
Dopamine
Epinephrine Adrenal medulla Norepinephrine

Dopamine -hydroxylase
Regulation of Norepinephrine Synthesis, Release and Metabolism:

Na+
Tyrosine

Tyrosine R

TH
Dopamine
Signal Dopa DD (DA)
DBH NE Uptake-1
ATP
NE
(-)
R
DBH Post-synaptic
Pre-synaptic Ca2+ NE
ATP
NE
NE
Cellular messengers
R
and effects
CO
M T
Ca 2+

Diffusion, Normetanephrine (NMN)

metabolism
Drug intervention -- Adrenergic transmission

: Stimulatory Tyrosine
: Inhibitory
Solid: Agonistic (Rate-limiting) TH Metyrosine
Dotted: Antagonistic
DopaDA
Reserpine
Adrenergic antagonists
Vesicle (DANE)
Phentolamine (-blocker)
Propranolol (-blocker) Amphetamine, tyramine,
ephedrine
Release
Adrenergic agonists Bretylium, guanethidine
(direct acting)
Cocaine
Isoproterenol Tricyclic antidepressants
Albuterol NE (e.g. imipramine)

Receptor Recapture
+ action by Uptake-1
Autonomic nervous system:

Receptor Functions 2/3/2016

ANS Receptors - Pharmacological Classification:
M1, M3, M5 (Gq coupled)
Muscarinic R
(mAChR)
M2, M4 (Gi coupled)
Cholinergic R
NM (neuromuscular, or muscle type ion channels)
Nicotinic R
(nAChR)
NN (neuronal,or ganglion type,ion channels)

1, 2( G protein coupled)
Adrenergic R
(all G protein 1, 2,
coupled)

Dopamine R D1, D2, D3, D4, D5 (G protein coupled)

(all G protein
coupled)
Other receptors (receptors for NANC transmitters,
e.g. nitric oxide, vasoactive intestinal peptide, neuropeptide Y)
 CNS Pre-ganglionic Ganglion Post-ganglionic

Cranial Effectors

Ach Ach Cardiac & smooth

Parasympathetic
muscles, gland cells,
(III,IIV,IX,& X) Nicotinic Muscarinic
nerve terminals
Ach NE
Sympathetic Cardiac & smooth
Adrenergic muscles, gland cells,
Nicotinic nerve terminals
()
Thoracolumbar

Ach Ach
Sympathetic
Sweat glands
Nicotinic Muscarinic

Ach D
Sympathetic Renal vascular
Nicotinic Dopaminergic smooth muscle
(D1)
Ach
Sympathetic (adrenal medulla)
Epi,NE Released into
Nicotinic blood
Sacral

Ach
Motor (somatic) Skeletal muscle
Nicotinic

Ach = acetylcholine D = dopamine Epi = epinephrine NE = norepinephrine

Adrenergic receptors
Classification of adrenergic receptors by agonist potency

-- NE Epi > Iso NE = norepinephrine

Epi = epinephrine
-- Iso > Epi > NE Iso = isoproterenol
OH
OH OH
CH CH
HO HO CH HO
CH2 CH2
HO CH2 HO
HO NH
NHCH3 NH2 Iso
Epi NE CH(CH3)2
Signaling properties of adrenergic receptors

Norepinephrine Norepinephrine Isoproterenol

Epinephrine Methyl NE Albuterol (2)
Phenylephrine Clonidine Dobutamine (1)
Agonist Agonist Agonist
1 2 1,2,3

Gq Gi Gs

Inositol phosphates cAMP cAMP

(IP3) Ca++
Calcium channels
Diacyl glycerol (DAG)
K+ conductance

Mostly excitatory Mostly inhibitory Mostly excitatory

 Gs and Gi proteins have different functions

Agonist Agonist
Beta1 receptor Alpha2 receptor(Gi)
(Gs)
AC
s i
s i

Gs = stimulatory G protein
Gi = inhibitory G protein
AC = adenylyl cyclase (converts ATP to cAMP which gives the effect)
Distribution & functions of adrenergic receptors:
1: postsynaptic effector cells, especially smooth muscle
Vasoconstriction, hepatic glycogenolysis

2: presynaptic adrenergic nerve terminals (autoreceptor), platelets, lipocytes,

smooth muscle.
Inhibition of transmitter release, platelet aggregation, relaxation of
GIT smooth muscle

1: postsynaptic effector cells: heart, lipocytes, brain, presynaptic adrenergic /

cholinergic terminals
Increased cardiac rate & force, relaxation of GIT smooth muscle
2: postsynaptic effector cells: smooth muscle, cardiac muscle
Bronchodilation, vasodilation, relaxation of visceral smooth muscle, hepatic
glycogenolysis
postsynaptic effector cells: lipocytes
Lipolysis
Dopaminergic receptors
 Tyrosine hydroxylase

3 1 CH HO CH2
2
TH HC COOH
HC COOH HO
HO
Phenylethanolamine- NH2
NH2 DOPA
N-methyl transferase Dopa decarboxylase
Tyrosine
DD (L-AAD) (L-amino acid
OH OH
decarboxylase)
HO CH HO CH
PNMT DBH HO CH2
CH2 CH2
HO HO
HO CH2
NHCH3 NH2
NH2
Dopamine

Epinephrine Norepinephrine Dopamine -hydroxylase

 Dopaminergic receptors in the periphery
Dopamine receptors play important roles in CNS.
Notably, dopamine neurotransmission is involved in
several diseases including Parkinsons disease,
schizophenia, and attention deficiency disorder.

There are 5 types of dopamine receptors (D1 D5). In periphery,

D1 dopamine receptors increase cAMP synthesis, mediate
renal vasodilation, and increased myocardial contractility.

Agonist D2,3,4 Agonist

D1,5

Gs Gi

cAMP in cardiac cAMP in smooth

muscle muscle
 IV dopamine in low dose promotes vasodilation of renal,
splanchnic, coronary,cerebral and other resistance
vessels via stimulation of D1 receptors,and
accummulation of cAMP in smooth muscle cells.
Activation of presynaptic D2 receptors inhibits release of
norepinephrine(similar to 2 receptors).
Dopamine also activates 1 receptors in heart.
In low doses, it decreases peripheral resistance, but at
high doses it stimulates vascular 1-adrenoceptors leading
to vasoconstriction including renal vasculature
Thus dopamine in high doses mimics action of epinephrine
 Receptor Regulation:
Adrenoceptor responses are not fixed and static.
Number and function of adrenoceptors on cell surface and responses may be
regulated by catecholamines themselves, other hormones and drugs, age and
diseases.These changes may modify the magnitude of physilogic response.
Examples of receptor regulaton is desensitization that occurs after exposure
to catecholamines or sympathomimetics for long periods. Other terms such
as tolerance, refractoriness and tachyphylaxis are also used to denote
desensitization
Desensitization may be due to transcriptional and translational changes in
the receptor protein level, or its migration to the cell surface, these occur
slowly over hours or days.Other mechanisms occur rapidly within minutes
due to receptor phosphorylation.
Cholinergic receptors
 Cholinergic receptors: Nicotinic(ion channels)

Nicotiana tabacum
(cultivated tobacco)

Nicotinic actions -- similar to those induced by nicotine.

Actions mediated by nicotinic cholinergic receptors include:
stimulation of all autonomic ganglia (NN receptors)
stimulation of voluntary muscle (NM receptors)
secretion of epinephrine from the adrenal medulla (N N)
 Nicotinic acetylcholine receptor: Function

Ligand-gated (Na+) ion channel

- an Ionotropic Receptor

Acetylcholine binds to the -subunits of the receptor making the

membrane more permeable to cations (sodium) and causing a local
depolarization. The local depolarization spreads to an action potential,
or leads to muscle contraction when summed with the action of other
receptors. The ion channel is open during the active state.
Nicotine in small doses stimulates autonomic ganglia and adrenal medulla.
When large doses are applied, the stimulatory effect is quickly followed by a
blockade of transmission(depolarization blokade).
 Nicotinic receptor antagonists
Competitive vs. depolarizing

Competitive Depolarizing
Physically blocks Binds and locks the receptor
Ach binding open
INHIBITOR
Cholinergic receptors: Muscarinic
Muscarinic actions -- reproduced by injection of
muscarine, from Amanita muscaria (mushroom).
Similar to those of parasympathetic stimulation
Multiple muscarinic cholinergic receptors
distributed in different tissues. Therefore, the
muscarinic actions are dependent on the
receptor types in different tissues and cells.
Neural/enteric (M1): CNS, ENS, gastric parietal cells (excitatory; Gq)
Cardiac (M2): atria & conducting tissue; presynaptic (inhibitory; Gi)
Glandular/endothelial (M3): exocrine glands, vessels (excitatory; Gq)
Neural (M4): CNS (inhibitory; Gi)
Neural (M5): CNS (excitatory; Gq)
(Sites of primary expression are listed; all are found in CNS)
 Muscarinic acetylcholine receptors
G Protein-Coupled Receptors (Metabotropic Receptors)

M1(enteric,neural) M2(cardiac)

M3(glandular,vascular) M4(CNS)
Agonist Agonist
M5(CNS)

Gq Gi
Mostly excitatory cAMP
CNS excitation Mostly inhibitory Ca2+ channel
IP3, DAG Gastric acid secretion Cardiac inhibition (Inhibition)
Gastrointestinal motility Postsynaptic
Intracellular Ca2+ inhibition K+ conductance
(Stimulation) Glandular secretion Neuronal inhibition (Slow IPSP)
Contraction of visceral
K+ conductance smooth muscle
(Depolarization) Vasodilation (via NO)
Clinical manifestation of excessive cholinergic effects

(DUMBELS)

D Defecation
U Urination
M Miosis
B Bradycardia
E Emesis
L Lacrimation
S Salivation
 Effects of muscarinic antagonists:
DRY AS A BONE, RED AS A BEET, MAD AS A
HATTER.
Decreased sweating, salivation and lacrimation
Reflex peripheral (cutaneous) vasodilation to
dissipate heat (hyperthermia)
CNS effects of muscarinic inhibition --
restlessness, delerium, hallucination
ALSO:
Bronchodilation
Tachycardia
Mydriasis (pupil dilation) and Cycloplegia (loss of focus)
GI and bladder atony leading to urine retension
Physiological Effects of ANS
Stimulation and Inhibition
Receptor distribution and effects in the autonomic nervous system:

Organ Sympathetic Receptor Parasympathetic Receptor

Heart Rate
SA node Rate M2
Force
Atrial muscle Force M2
Automaticity
AV node Conduction velocity M2
AV block
Automaticity
Ventricular muscle
Force
Blood vessels
Arterioles Contraction
Coronary Relaxation
Skeletal muscle Contraction M3
Viscera Contraction
Skin Contraction
Brain Contraction Relaxation M3
Erectile tissue Contraction Relaxation M3
Salivary gland Contraction
Veins Relaxation


(Continued, next page)
 Organ Sympathetic Receptor Parasympathetic Receptor
Viscera
Bronchiolar SMC Relaxation Contraction M3
Glands Secretion M3
GI track
Smooth muscle Motility Motility M3
Sphincters Contraction Relaxation M3
Glands Secretion M3
Uterus M1
Contraction Gastric acid secretion
Relaxation Variable

Skin
Pilomotor SMC Contraction (piloerection)

Salivary glands Secretion Secretion M3

Lacrimal glands Secretion M3
Kidney Renin release
Liver Glycogenolysis
Gluconeogenesis

Fat Lipolysis

From: Rang et al. Pharmacology, 6th Ed. p. 169. Also, see Katzung, Basic & Clinical Pharmacology, 10th Ed. p.86.
Cardiovascular Pharmacology
(Blood Pressure)
Cardiovascular effects of IV infusion of epinephrine, norepinephrine,& isoproterenol in man:
Norepinephrine (predominantly -agonist) causes vasoconstriction and increased systolic
and diastolic BP, with a reflex bradycardia.
Isoproterenol (-agonist) is a vasodilator, but strongly increases cardiac force and rate.Mean
arterial pressure falls.
Epinephrine combines both actions,since it stimulates both receptors equally).
 Sir Henry Hallett Dale
(Nobel laureate, 1936)

(Arterial pressure of an
anesthetized cat was
Two kinds of effects produced by Ach. measured)
A. Ach causes a fall in BP due to vasodilation.
B. A larger dose of Ach also produces bradycardia, further reducing BP.
C. Atropine blocks the effect of Ach in lowering BP.
D. Still under the influence of atropine, a much larger dose of Ach causes a rise in BP& tachycardia.
A, B: Muscarinic effects of Ach (M3, M2)
C: Muscarinic antagonistic effect (M)
D. Stimulation of sympathetic ganglia (NN)
 Mechanism of action of Ach
Ach modifies organ function by two mechanisms:
-Ach released from parasympathetic nerves binds to
muscarinic receptors on effector cells and alter their
function.
-Released Ach binds to muscarinic receptors on nerve
terminals and inhibits release of neurotransmitters.
All muscarinic receptors are G protein coupled
Binding of muscarinic agonists to receptors activates IP3
and DAG cascade.
 DAG opens smooth muscle calcium ions channels.
IP3 releases Ca++ from endoplasmic & sarcoplasmic
reticulum.
Muscarinic agonists increase cellular cGMP concentration
Activation of muscarinic receptors also increases potassium
flux across cardiac cell membranes and decreases it in ganglia
and smooth muscle cells.This effect is mediated by the
binding of an activated G protein subunit to the channel
Muscarinic agonists attenuate activation of adenylylcyclase
and decrease cAMP levels,this reduces physiological response
to stimulatory hormones
 Cholinergic action(molecular mechanisms):
Intracellular signalling triggered by Ach in smooth muscle
(contraction) main players:m3,Gq,PLC,IP3,DAG,Ca++
Nitric oxide (NO) signaling pathway for SMC relaxation

Second
messenger

Inhibition of PDE causes sustained increase in level

of cGMP that maintains smooth muscle relaxation.

Sildenafil (Viagra)
is an inhibitor for
PDE 5.
Intracellular signalling triggered by Ach in heart muscle: results in
bradycardia. Main molecular players: M2, Gi, adenylylcyclase
Pulmonary Pharmacology
(Asthma and COPD)
Ocular Pharmacology
(Glaucoma)
 Cholinergic effects:
Contraction of pupillary constrictor muscle
-- miosis
Contraction of ciliary muscle - bulge of lens
-- near vision, outflow of aqueous humor
Pupillary dilator muscle ()
Pupillary constrictor muscle
(M3)
Adrenergic effects:
Contraction of pupillary dilator muscle
-- mydriasis
Stimulation of ciliary epithelium
-- production of aqueous humor

Trabecular meshwork
(opened by pilocarpine)

Lens

(M3)
Secretion of aqueous humor ()