Basics of Neuropharmacology: Neurotransmitters I

Dr. Demetrio Sierra Mercado

Catedrático Auxiliar
Departamento de Anatomía y Neurobiología
Universidad de Puerto Rico Recinto de Ciencias Médicas
23-marzo 2017 1
Audio and/or video recording of lecture is not permitted
 General Goal of Neuropharmacology

Apply information about drugs and their mechanisms of

action to develop safer, more effective treatment for nervous
system abnormalities.

Objectives
Further understand:

•  Some neurotransmitters and some of their functions

•  The life-cycle of a neurotransmitter

•  How to manipulate the function of a neurotransmitter

 What is a neurotransmitter?

-  Chemical substance synthesized in a neuron

-  Released at a synapse following depolarization

-  Binds to a postsynaptic receptor and/or a presynaptic

terminal to elicit a response

3
 Synaptic Transmission
100 billion neurons in CNS.
86 billion neurons in CNS (Azevedo et al., J Comp Neurol, 2009).

Chemical synaptic transmission.

Excitation: increases probability that neuron membrane potential
reaches threshold and fires action potential.
Inhibition: decreases probability
Emem (mV)

-60 Threshold -60

-70 EResting -70

E E E I E
 Changes in synaptic transmission
Drugs, patterns of stimulation, disease, toxins can affect synapses
either by altering the amount of transmitter released or by altering
the effect of the transmitter on the postsynaptic receptors
 Basic Terminology
Agonist
Chemical that binds and activates a receptor

Antagonist
Chemical that binds and blocks a receptor
 Types of Ion Channels in CNS

Fast neurotransmitters
(voltage- and ligand-gated)

Vs

Slow neurotransmitters
or neuromodulators

Basic & Clinical Pharmacology,

Katzung, McGraw Hill, 2001, p353.

Synaptic speed determined by receptor, not neurotransmitter

Postsynaptic Mechanisms: Basis of Excitation and Inhibition

Lichtman
 The synaptic cleft

Presynaptic
50 nm

........
. . ..
.
. .
20nm

Postsynaptic

Time for transmitter to diffuse across cleft: < 50µs

Time for transmitter to diffuse out of the cleft: ~ 1ms
Mechanisms needed to stop neurotransmitter action Lichtman
 The lifecycle of neurotransmitter
Released neurotransmitter can be:
1)  Taken up by the presynaptic nerve terminal: Reuptake
2)  Broken down by synaptic cleft enzymes
3)  Taken up by glia
4)  Diffuse
5)  Sometimes combinations: neurotransmitter may be cleaved by enzyme in cleft and
components taken up to make more

Presynaptic

reuptake

glial uptake enzymatic

destruction diffusion away

bind and unbind

to neurotransmitter
Postsynaptic receptor

Lichtman
 Clinical Application
1)  Reuptake inhibitors
2)  Enzyme inhibitors

Presynaptic

reuptake

glial uptake enzymatic

destruction diffusion away

bind and unbind

to neurotransmitter
Postsynaptic receptor
 Sites of Drug Action in CNS

EXAMPLES
1 1) Metyrosine
2 2) Reserpine
3
3) Lidocaine
4) Amphetamine
4
5) Atropine
8
7 6) Tacrine
7) Clonidine

5 6 8) Imipramine; TCA

Pharmacology, Brenner, WB Saunders (2000), p177.

 Vesicles
Synaptic vesicles are the
structural basis of quanta: they
contain neurotransmitter which
they release by fusion with the
presynaptic membrane
(exocytosis).

Synaptic vesicle
Neurotransmitter
Release
apparatus

Fusion
(Exocytosis)
Neurotransmitter
release
Sanes
 The SNARE complex: Synaptic vesicle proteins
Interactions between SNARE proteins on the vesicle and on the
presynaptic membrane, modulated by calcium, lead to vesicle fusion.

Takamori et al.,
Cell, 2006 Sanes
G protein-coupled receptors (GPCRs)

Extracellular domain binds transmitter,

hormone, other molecule
~2000 GPCR genes each with different
binding preference….Nearly 10% of all
genes!
Intracellular domain binds a set of three
proteins - α, β,and γ- together called G
proteins because they bind GTP
Binding of ligand outside affects G
proteins inside. For example, α can split
from β and γ, move along membrane,
interact with other proteins, and activate
or inactivate them.
 Second messengers
Frequently, the activated proteins are
enzymes, which generate small
molecules that affect channels (or
something else). These are called
"second messengers.”

A particularly important second

Cyclic AMP, a
second messenger messenger is “cyclic AMP,” which is
generated from the energy source,
ATP, by a G protein-activated
ATP → cAMP + PPi
→ enzyme, adenylyl cyclase.

CyclicAMP activates another

Inactive → active protein kinase A
→ enzyme, protein kinase A, which can
activate many proteins, including
channels.
Inactive channel → Channel-P
→
(active)
 Versatility of GPCR signaling
A. Amplification Few molecules exert
a big effect.
 Versatility of GPCR signaling
A. Amplification Few molecules exert
a big effect.
B. Multiple effects on a single
target. Different GPCRs exert
different effects on the same
enzyme.

Gs Gi

Used in the autonomic system…allows norepinephrine

and acetylcholine to exert opposite effectd (“fight or
flight” and “rest and digest” respectively) on the same
processes.
 Versatility of GPCR signaling
A. Amplification Few molecules exert
a big effect.
B. Multiple effects on a single
target. Different GPCRs exert
different effects on the same
enzyme.
C. Multiple targets. Messengers
can affect channels in the
membrane, metabolites in the
cytoplasm. They can also affect
gene expression in the nucleus,
leading to long-lasting changes
(like memory???)
 Drugs Can Interact with Multiple Receptors

Tricyclic Antidepressant

Histamine
NE Reuptake Muscarinic Receptor
Transporter Receptor

Decrease Antimuscarinic Sedation

Depression Effects
 Adaptation to Chronic Drug Treatments
Antagonist
Compensation ⇒ up-regulation
Example
Chronic Antipsychotics which block dopamine receptors
induces the production of more dopamine receptors &
produces hyperkinetic disorders.
Also there is a delayed onset of therapeutic effects.

Agonist
Compensation ⇒ down-regulation
Example
There is a delayed onset of therapeutic effects of
antidepressants which block serotonin uptake.
Probably involves changes in rates of receptor synthesis.

Pharmacology, Brenner, WB Saunders (2000), p184.

 Many Classes of Neurotransmitters
(each with different functions)
Amino acids Purines
Excitatory Adenosine
Glutamate ATP
Inhibitory
γ-amino-butyric Neuropeptides
acid (GABA) Endorphins (opiods)
Glycine Substance P

Biogenic Amines NO Generally not stored in

synaptic vesicles;
Acetylcholine Generated in
Catecholamines Endocannabinoids response to increases
anandamide in intracellular Ca2+
Norepinephrine
and freely diffuse out
Dopamine of neurons
Serotonin
Histamine
We will focus on one class for now…
 Glutamate Receptors

Major excitatory transmitter at many synapses

Cortico-cortical
Thalamocortical: Cortico-thalamic
Cortico-striatal (Extrapyramidal pathway)
Cortico-spinal (Pyramidal pathway)
1º sensory neurons

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 Glutamate’s Lifecycle

Glu H+
2 1
Glu Glu 1) Synthesis from
Glucose via Krebs cycle
Glu 2 Na+
3 2) Vesicular storage via Glu
G Glu 5 transporters
Glu
Glu Glu
3) Ca2+-dependent release
Ca2+
Na+ Na+ Na+
Glu Glu 4) Activates receptors
4
G
5) Reuptake into nerve terminals
NMDA AMPA Kainate & astrocytes
mGluR Na+ Ca2+ Glutamine by glutaminase
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 Glutamate Receptor Subtypes

Fast Transmission Slow Transmission

1) NMDA 4) Metabotropic (G-protein)

Agonist: N-methyl-D-aspartate G-protein coupled
↓ cAMP
Cation Channel: Na+, Ca2+, K+
↑ IP3 & Ca2+

2) AMPA
Agonist: AMPA Postsynaptic excitation
Cation Channel: Na+, Ca 2+ , K+ Presynaptic inhibitory
Autoreceptors

3) Kainate
Agonist: kainate
Cation Channel: Na+, Ca , K+
2+

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 Both NMDA & AMPA Receptors @ Most Glutamatergic Synapses

AMPAR NMDAR

Lissin et al.; J. Neurosci.;19(4); 1999.

Why have both NMDA & AMPA receptors at single synapses?

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Mg2+ Block of NMDA Receptors at resting membrane potential

What force expels Mg2+?

AMPA receptors provide

initial depolarization
needed to expel Mg2+
from NMDA receptors.

Ca2+ influx through NMDA

receptors activates
many intracellular
enzymes.

28
Neuroscience Exploring the Brain;Bear, Connors, & Paradiso: Lippincott Williams & Wilkins; 2001; p 153.
 OJO: NMDA Receptors also require Glycine

Important because activation of some receptors may

29
require more than one chemical (co-agonist)
 Importance of glutamate receptors (GluRs)

1) Learning & Memory formation

Some memories are stored by enhancing glutamatergic
synaptic transmission. Often requires influx of Ca2+
through NMDA receptors.

Glutamate and NMDA receptors are necessary for synapse 30

plasticity (e.g. thought to occur in learning).
 Some types of learning require NMDA and AMPA
Pairing a neutral
stimulus (a tone)
and an aversive one
(foot shock)
Foot shock
Eventually the tone
alone gives the
animal all the
emotional
expression of fear
(high blood
pressure and
Tone
“freezing”)

Long-term potentation
(Hebbian synaptic
mechanism?)
Lichtman
• NMDA receptors are ligand (glutamate
and voltage gated releasing)
Strong pain
• NMDA receptors blocked by
input
Mg2+; glutamate opens Ca2+
channel only when the
postsynaptic membrane is AMPA R
already depolarized, in this
case by the pain input
Depolarize post-
synaptic cell
• Thus only weak
inputs that are
synchronously active with
the pain input allow local
Ca++ entry
NMDA R
• The Ca++ entry causes new Postsynaptic AMPA R
AMPA receptors to insert into
NMDA Rs
the postsynaptic site -
potentiating the formerly
weak synapse (e.g. long-
term potentiation LTP)
Lichtman
• NMDA receptors are ligand (glutamate
and voltage gated releasing)
Strong pain
• NMDA receptors blocked by
input
Mg2+; glutamate opens Ca2+
channel only when the
postsynaptic membrane is AMPA R
already depolarized, in this
case by the pain input
Depolarize post-
synaptic cell
• Thus only weak
inputs that are
synchronously active with
the pain input allow local
Ca++ entry
NMDA R
• The Ca++ entry causes new Postsynaptic AMPA R
AMPA receptors to insert into
NMDA Rs
the postsynaptic site -
potentiating the formerly Synchronously
weak synapse (e.g. long-
term potentiation LTP)
STRONG
active weak
Lichtman
tone input
¿Other Clinical Applications of NMDA Receptors and
Plasticity?

34
 Importance of GluRs

2) Epilepsy: imbalance in excitation and inhibition

Some antiepileptics block glutamate receptors.

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 Importance of GluRs

3) Excitotoxicity
Excessive stimulation ⇒ excessive Ca2+ influx ⇒ neuronal
damage ⇒ neurodegeneration
Stroke
Amyotrophic lateral sclerosis (ALS; Lou Gehrig’s disease)
Multiple sclerosis

Development of NMDA receptor blockers to prevent

excitotoxicity
Memantine (Namenda)

36
 Importance of GluRs continued
4) Dissociative anesthesia (amnesia, catatonia, and
analgesia)
Ketamine blocks NMDA receptors.

37
 Importance of GluRs continued

5) Drug abuse
PCP (angle dust) is NMDA receptor antagonist. Reducing
NMDA receptor activation can cause hallucinations.

38
Glutamate and schizophrenia

(Gordon et al., 2010) review on Belforte et al., 39

 Classes of Neurotransmitters
Amino acids
Excitatory
Glutamate

Inhibitory
GABA*
Glycine*

40
 γ-aminobutyric acid (GABA)

Major inhibitory neurotransmitter in Brain

2 groups of neurons
Interneurons:
local circuit neurons (ie axons do not extend to other brain areas)
neocortex, thalamus, striatum, hippocampus, cerebellum, spinal cord

Projecting neurons
Striatum ⇒ globus pallidus ⇒ thalamus/ substantia nigra

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 GABA’s Lifecycle

GABA
H+ Glu
2 1 1) Synthesis from
GABA GABA Glutamate by glutamic acid
decarboxylase (GAD)

GABA 2 Na+ Ojo: synthesized from amino

3
G GABA 5 acid
GABA 2) Vesicular storage via GABA
GABA GABA
transporters

Cl- Cl- 3) Ca2+-dependent release

GABA GABA

4
4) Activates receptors
G
Cl- 5) Reuptake into nerve terminals
GABAB GABAA & astrocytes

6) Vesicular storage or 42
metabolism
 GABAA Receptors

Fast transmission
↑ Cl- channel
hyperpolarizes
neuron

ECl= -65 mV

43
Neuroscience Exploring the Brain;Bear, Connors, & Paradiso: Lippincott Williams & Wilkins; 2001; p 116.
 GABAA receptor has many modulatory sites

Allosteric enhancers

Disorders including Epilepsy, Anxiety Disorders, Insomnia, alcohol

withdrawal can be treated by enhancing the effect of GABA at
GABAA receptors (ie more Cl- influx) using benzodiazepines such
as diazepam, & barbiturates such as phenobarbital.
44
Principles of Neural Science; 4th Ed; Kandel, Schwartz, Jessell; McGraw Hill;p338.
 GABAB Receptors

1. Postsynaptic ↑ K+; inhibitory

Slow transmission
GABA G-protein coupled

GIRK
βγ + β
αi γ

GDP GTP

2. Presynaptic ↓ Ca2+; inhibitory

Baclofen is an agonist of GABAB receptors and reduces muscle

spasms by increasing inhibition in spinal cord.
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 Glycine
Spinal interneurons
Some brain stem interneurons
Glycine receptors
-Ligand-gated ion channels
-Fast inhibitory transmission by
allowing Cl- influx
Hyperekplexia
-Mutations in glycine receptor
-Not enough Cl- passes
-insufficient inhibition in spinal cord
and generalized stiffness and
excessive startle reflexes.
46
 Glycine

Strychnine Opisthotonos
-antagonist of glycine receptors

-used to kill rodents causing muscular

convulsions and eventually death
through asphyxia by blocking glycine
receptors in spinal cord

Tetanus toxins
-Cleaves vesicle proteins
preventing release of glycine and
GABA in spinal cord

-Cause muscle rigidity

47
 Glycine’s Lifecycle

Glycine
H+ serine 1) Synthesis from
2 1
Serine
Glycine Glycine
2) Vesicular storage via Glycine
transporters
Glycine 2 Na+
3
Glycine 5
3) Ca2+-dependent release
Glycine
Glycine
4) Activates receptors
Cl- Cl-
Glycine
5) Reuptake into nerve terminals &
4 astrocytes
Cl-
48
 Briefly, one more class of neurotransmitter
Purines: Adenosine Receptors
ATP (Adenosine Triphosphate)
Metabolized to Adenosine upon release

Clinical importance
Adenosine has inhibitory effect on CNS
What happens if Adenosine Receptors are blocked?
Clinical importance
Xanthines (caffeine, theophylline, theobromine) block adenosine
receptors producing arousal.