What is an adrenergic response?

The fight or flight response

 Lecture outline

• Catecholamine
- Synthesis
- Storage/release
- Termination
Reuptake/Metabolism/Drug interventions
- Receptor classification
Locale/Physiological effects
- Molecular Mechanisms
- Factors determining receptor activity
Nerve Endings
 Tyrosine Hydroxylase

• Tyrosine Hydroxylase is rate limiting

• An increase in sympathetic n.s. activity leads to:
  in NE release
  in NE Pool and release of feedback inhibition
  in Feedback inhibition (NE, PKA, PKC)
  in TH activity
  in NE synthesis
 Dopamine β-hydroxylase

• Found exclusively in vesicles that store NE or EPI

• Lack of substrate specificity
• Converts a broad range of phenylethylamines
(PEA’s) to β-hydroxylated products that can be
stored.
 Storage and Release

Sympathetic Nervous System (SNS) activation generates nerve

action potentials and influx of Ca++ at the nerve terminus that
stimulates

-NE and EPI (100:1) release from the presynaptic postganglionic

neuron into the synaptic compartment: mechanism is termed Stimulus
Secretion-Coupling (or exocytosis).

-EPI and NE release (85:15) from the Adrenal Medulla Chromaffin

Granules (remember that stimulation of preganglionic nerves cause release
of Ach that activates nicotinic AChR on the adrenals). Mechanism of
release is similar to stimulus secretion coupling and catecholamines are
emptied into the blood stream.

Note: ATP is present in vesicles at high concentration and plays

significant but ill understood roles following activation of purinergic
receptors (there are many).
STORAGE
INTERACTION WITH EFFECTOR CELL
 Termination of NE Action

• Reuptake into adrenergic neurons: Two step

uptake; uptake I (PM) and vesicular uptake
• NE feedback inhibition on α2-adrenergic
receptors; blocks release of NE
• Diffusion from site of action and uptake at non-
neuronal sites (uptake II)
• Inactivation by catechol-o-methyl transferase
(COMT) and monoamine oxidase (MAO)
• Desensitization of NE action on the
effector cell
Inhibitory feedback loop blocks
NE release through 2-adrenergic
receptors
TERMINATION OF ACTION

Vesicular
uptake Uptake 2

Uptake1

diffusion
 NE transporter (uptake 1)
• Requires Na+ and ATP
• Active process
• Steriospecific (l-NE); Km about 1.0 M
• Works against a concentration gradient
• Many compounds are good substrates (EPI, NE,
DA, tyramine (TA), PEA, guanethidine,
amphetamine
REUPTAKE BLOCKERS
 NE Uptake 1 Inhibitors potentiate
sympathetic stimulation
• Cocaine:
-competitive inhibitor of NE uptake
-relatively non-specific; blocks serotonin
(5HT) and dopamine reuptake
• Tricyclic antidepressants
desipramine (NE selective)
imipramine (tofranil)
amitryptyline (elavil), relatively 5HT
selective
floxetine (Prozac), blocks DA, NE and
serotonin uptake
 Vesicular uptake system and
inhibitors
• Vesicular uptake an active process requiring Mg and ATP
• Uptake of catechols is non-specific; works for NE, EPI,
DA as well;
• Inhibitor: Reserpine, an alkaloid from the roots of
Rauwolfia serpentina originally used as an anti-psychotic
– Blocks the ability of vesicles to take up and store
biogenic amines NE, dopamine, 5HT
– Essentially irreversible in action
– Once used to treat hypertension
Guanethidine: somewhat similar drug is taken up and
concentrated in nerve endings and interferes with
stimulus secretion coupling-bad side effects
 Monoamine Oxidase and Inhibitors
• Converts catechols to aldehydes non-selectively
• Found in sympathetic nerve endings, liver and intestinal
mucosa
• Protects body against phenylethylamines in diet like
tyramine (found in beer, cheese, wines, yeast, soy sauce)
• Inhibitors include:
-clorgyline for MAO-A (5HT selective)
-selegiline for MAO-B (dopamine selective
therapy for Parkinson’s)
isocarboxazid-rarely used
Question: What happens if a patient on a MAO inhibitor eats a
large quantity of PEA’s?
 COMT Inhibitors
• Entacapone and Tolcapone
– USE: As adjunct therapy with DOPA (l-dopa)
in Parkinson’s Disease
– MOA: inhibits COMT and prolongs DOPA
action
 Excretion of Catecholamines in Humans
Amount in Urine/ 24 hours

mg
3-Methoxy-4-hydroxymandelic acid(VMA) 3.8
3-Methoxy-4-hydroxyphenylethylene glycol (MOPEG) 1.5
Normetanephrine 0.2
Metanephrine 0.1
Norepinephrine 0.03
Epinephrine 0.005
 Drugs can Enhance Adrenergic
Neurotransmission by Different Mechanisms

Mechanisms Example
• Tyramine : displacement
1. Increase release of NE
• Yohimbine : blocks 2 receptors

2. Mimic Interaction with
postjunctional receptors
3. Inhibit termination of NE
• Phenylephrine : 1 agonist
• Dobutamine : 1 agonist
• Cocaine, desipramine : blocks
neuronal uptake
• Pargyline, clorgyline, selegiline :
MAO inhibitors
Drugs Can Inhibit Adrenergic Neurotransmission
by Different Mechanisms
Mechanisms Example
Inhibit synthesis of NE  - Methyltyrosine : inhibits tyrosine
hydroxylase

Inhibit storage of NE Reserpine : blocks vesicle transport

Inhibit release of NE
Inhibit NE interactions with
postjunctional receptors
system
Guanethidine : blocks Ca++ dependent
exocytosis
Clonidine : agonist at presynaptic 2
receptors
Prazosin : 1 antagonist
Metoprolol : 1 antagonist
 Classification of Adrenergic
Receptors
• Discovery
• Subtypes of alphas and betas
• Functions
• Localization
 Discovery/Classification of Adrenergic
Receptors
• Originally based on complex and often
paradoxical physiological responses to NE, EPI
and ISO: led to concept of  and  receptors
• Synthesis of new adrenergic agonists and
antagonists allowed distinction of  subtypes
first, and then  subtypes
• Cloning further clarified the subtypes and allowed
their independent study after individual expression
in cells grown in vitro
Comparison of agonist potency on - and -
adrenergic receptors

: EPI>NE>>ISO
: ISO>EPI>>NE

1: ISO>EPI = NE
2: ISO>EPI>>NE
 Adrenergic Receptor
Classification
 -Adrenergic  -adrenergic
1 1A
2 1B
3 1D

2A
2B
2C
 Adrenergic Receptors
Alpha () receptors are generally excitatory
(constriction/contraction) except in gut
Mechanism 1 :  Ca++

Beta () receptors are inhibitory (e.g. 2 lung

relaxation;) or stimulatory (heart, liver and
fat cells)
Mechanism:  cAMP
“Fight or Flight”

FAT

Epi
NE
 Factors determining the actions of
catecholamines on ARs
• Reuptake • Desensitization
• Catabolism • Down regulation
• Density of receptors • Removal of 2nd
• Relative proportion of messenger
and locale of • Pharmacogenetics
adrenergic receptors • Reflexes
• Potency of agonist
• Efficiency of agonist
 -Adrenergic Receptors
1 2
Heart (stimulation) Arterioles of liver and
Fat ( lipolysis; 3 also) skeletal muscle (relax)
Kidney (↑ renin release) Coronary and cerebral
vessels (relax)
Amylase secretion (thick) Bronchioles (relax)
Uterine smooth muscle
(relax)
Eye: Increase in aqueous
humor; relaxes ciliary Metabolic (liver, muscles)
muscle Skeletal muscle
(↑contraction)
Mast Cell granule release
(inhibition)
 -Adrenergic Receptors
 1-receptors
– Constriction of smooth
muscle
• Arterioles of skin, mucosa,
abdominal viscera
• arterioles of liver and
skeletal muscle
• Coronary/cerebral vessels
• Veins
• bladder sphincter, seminal
tract, iris radial SM
Relaxation intestinal wall
SM
 2-receptors
– Presynaptic inhibition of
NE release
– Constriction in arterioles of
kidney, abdominal viscera
and veins
– Platelet aggregations
– Decreased secretion of
insulin
Different blood vessels contain different
densities of / receptors

Renal, skin vessels, mucosa mainly 1

Abdominal viscera, /

Skeletal muscle vessels, mainly 2

(*Think about blood flow in Fight or

Flight)
REDISTRIBUTION OF BLOOD FLOW
 ANS AMAZING FACT

Cardiovascular reflexes may OVERRIDE the

direct effects of a drug

Example: slowing of heart after injection of

Norepinephrine
Homeostatic reflexes complicate drug effects
Example: what are the consequences of slowly infusing NE into a patient?

NE directly produces TACHYCARDIA and vasoconstriction.

However, the net effect of NE infusion is often a large increase in TPR,
a moderate increase in MAP, and BRADYCARDIA. Why?
 Pharmacogenetics

 2-adrenergic receptor is highly polymorphic (altered

amino acid sequence)
• Altered structure may affect agonist activation or
desensitization
– E.g.; Thr  Ile 164 impairs agonist activation

– Patients with this variant of the 2-adrenergic receptor

appear to have increased risk of dying from congestive heart
failure and go to the front of the line for transplant
Mechanisms of Desensitization of the β2-Adrenergic
Receptor
Homologous PKA-mediated
Heterologous GRK/Arrestin- mediated

1.Agonist binding (NE or EPI) rapidly activates adenylyl

cyclase and PKA and PKA phosphorylates the β2AR
2.Agonist binding to the β2AR also rapidly activates GRKs
3.GRK phosphorylation of the β2AR leads to Arrestin binding
and uncoupling
3. Arrestin binding leads to clathrin coated pit internalization
of the β2AR
4. Internalized receptor is recycled to the plasma membrane
(dephosphorylation occurs both internally and at the plasma
membrane resenstizing the β2AR )
5. A small fraction of the β2AR is downregulated (degraded)
slowly (t1/2 = 3-4 hrs)
Billington and Penn, Respir. Res 2003,4:2