Psych 215

Lecture 5a: Serotonin

 Lecture 5a: 5-HT Objectives
• Explain the derivation of the name “serotonin”
• Describe the chemical features of 5-HT that distinguish it from catecholamines
• Describe the two general 5-HT systems in the brain in terms of their projections
• Distinguish between the projections of the D and M ascending 5-HT systems based on
morphology and sensitivity to toxins
• Outline the pathway through which 5-HT is made from tryptophan, indicate which enzyme is the
rate-limiting step in 5-HT synthesis and highlight known regulatory mechanisms
• Explain why altering dietary tryptophan levels can affect 5-HT levels
• Describe the mechanisms of action of PCPA, fenfluramine, methamphetamine, MDMA (ecstasy),
LSD, reserpine, ondansetron, fluoxetine, paroxetine and sertraline as they relate to 5-HT
transmission
• Explain how one could demonstrate that serotonin releasers such as fenfluramine act on non-
vesicular stores of 5-HT
• Explain the effects of agonists and antagonists at 5-HT1A and/or 5-HT1B/1D receptors upon
extracellular levels of 5-HT
• Describe the intracellular signaling pathway for 5-HT1 receptors and relate this signaling to their
role in regulating 5-HT release.
• Describe the intracellular signaling pathway for 5-HT2 receptors and relate this signaling to their
hallucinogenic profile.
• Describe some of the oddities observed regarding the regulation of 5-HT2 receptor expression by
agonists and antagonists
• Distinguish 5-HT3 receptors from other 5-HT receptors in terms of structure, function, and
localization in brain
• Explain the neuronal consequences of the repeated administration of 5-HT releasers and predict
which brain regions/behaviors would be most likely affected
Indolamine-Serotonin
catecholamines

Isolated from blood and induced muscle contraction in heart=“serum” +

“tone”=serotonin
Isolated from intestinal mucosa and induced muscle
contraction=enteramine (secreted by enterochromaffin cells in GI tract)
-90% in GI; 8%-10% in blood platelets & 1-2% in brain (but several 100,
000 5-HT neurons in your brain)
-cannot cross the BBB
5-HT projections in brain

Nine 5-HT containing cell body groups located in the brain stem (B1-9)
Two systems: caudal versus rostral systems

Caudal system: B1-4; medulla and central pons; project down spinal cord in
several pathways (ventral horn, dorsal horn, preganglionic sympathetic cellsÆrole
in sensory, motor and autonomic functioning)
Rostral system: B5-9; rostral pons and mesencephalon (dorsal and medial
raphe, B6-B8, provide 80% of forebrain 5-HT)Æinnervate cortex, basal ganglia,
habenula, thalamus, hypothalamus, limbic system
Synthesis Tryptophan
hydroxylase=rate-limiting
step
-made in cell bodies and
transported to terminal for
5-HT synthesis
-tryptophan crosses BBB
and can be regulated by
diet
-regulated by PKA, CaMKII
-gene regulated by
cAMP/CREB although no
CRE site
-no end-product feedback
-PCPA=irreversible
antagonist (2 weeks)

AADC=very rapid enzyme

(same as for converting
DOPA to dopamine; found
in terminal)
 5-HT metabolism:
2 processes:
1-deamination & 2-oxidation

1. MAO-A: oxidative
deamination of catecholamines
-enzyme localized in synapse
as well as in terminal; cytosolic
MAO responsible for majority of
break-down after re-uptake

2. Aldehyde dehydrogenase:
Æ5-HIAA which diffuses out of
terminals and into CSF
-spinal tap for index of 5-HT
release/reuptake

MAO-A
inhibitors
5-HT projections in brain
2 types of 5-HT fibers in the brain:

1-Thin; very numerous; many varicosities

the “D” system (from dorsal raphe)
More vulnerable to toxins (e.g., PCA & MDMA)
Make less direct synaptic contact
(varicosities)Æmodulatory effects upon frontal
cortex, striatal structures

2-Thick, large varicosities

the “M” system (from medial raphe)
Make more conventional synaptic contacts
Æhippocampus and septum
Terminals & Release:
Resirpine: Blocks VMAT
Depletes 5-HT stores

5-HT releasers: PCA and

fenfluramine, MDMA,
methamphetamine and at very high
doses amphetamine

-non-vesicular mechanism of releasing action because not blocked by reserpine

-involves transporter because blocked by re-uptake inhibitors
 5-HT toxicity:
“D” fibers are particularly sensitive to releaser-type neurotoxins:

Terminal degeneration; cell body sparing; get both M and

D systems; Regrowth; Super-sensitivity to agonists

5,7-DHT MDMA or fenfluramine: Selective for D system;

Regrowth?

Releaser Releaser
Auto-inhibition: Gi-coupled 5-HT receptors

-cell body level: 5-HT1A

-terminal level: 5-HT1B/1D
 Re-uptake:

SSRI

Non-selective re-uptake inhibitors

-bind DAT, NET and SERT
Paroxetine=Paxil, Aropax, Oxetine, Aroxat, Cebrilin, Deroxat, Motivan,
Paroxetina, Optipar, Paroxat, Pondera, Seroxat, Posivil, Pexot, Paraxyle,
Plasare, Paradise CR

Sertraline=Zoloft, Sertralin, Lustral, Apo-Sertral, Asentra, Gladem, Serlift,

Stimuloton, Xydep, Serlain, Concorz

Fluoxetine=Prozac
5-HT receptor subtypes:
5-HT1 receptors:
Anti-depressant Rx’s can reduce 5-HT1A and 5-HT1D/1B autoreceptor function
5-HT2 receptors:
Antagonists Agonists

AgonistsÆvery rapid down-regulation; chronic state of super-

sensitivity???
AntagonistsÆdown-regulation of receptor function???inverse agonists?
 5-HT3
receptors:

Antagonists
 MDMA @ somatodendritic
methamphetamine */* 5-HT1A receptors
fenfluramine
PCPA

reserpine

*/*
MAO-A I’s

MDMA
SSRI methamphetamine
fenfluramine

LSD/anti-psychotics */* */*

ondansetron
Lecture 5b: Acetylcholine
 Lecture 5b: ACh Objectives
• Describe how Acetylcholine (ACh) is synthesized.
• Describe the sources for the Acetyl CoA and Choline used for ACh
synthesis and discuss what limits ACh synthesis in brain.
• Explain the role of acetylcholinesterase (AChE) in terminating ACh activity,
describe the factors regulating it and describe the neurochemical and
behavioral effects of inhibition
• Compare and contrast muscarinic and nicotinic receptors in terms of their
effectors systems and distribution and provide examples of agonists and
antagonists of each
• Explain the structure of the nicotinic receptor and how it can give rise to at
least 9 different subtypes.
• Explain and illustrate how drugs may interfere with ACh activity by
effecting its synthesis, release, degradation or receptor activity.
• Explain why atropine can ameliorate the effects of AChE inhibitors.

Lecture 5c: Histamine Objectives

• Describe how histamine can be synthesized
• Compare and contrast the 3 histamine receptors and explain (1) why H1
antagonists make you drowsy and (2) why H2 compounds cannot be used
currently to treat neuropsychiatric disorders
 Acetylcholine (ACh)

The first neurotransmitter to be discovered.

Otto Loewi, 1921

– All muscular movement is accomplished

by the release of acetylcholine.

– Appears to be involved in regulating REM

sleep, perceptual learning, and memory.
 ACh Synthesis
• ACh is synthesized in the cytoplasm from
Acetyl CoA and choline in a reaction
catalyzed by the enzyme choline
acetyltransferase (ChAT).
Sources of Acetyl CoA
and choline
 Choline
• Obtained from foods such as egg yolks, kidney, liver, seeds, and
various vegetables. It is also naturally produced by the liver.

• Free Cholie and lipid-bound choline circulating in blood plasma readily

crosses the BBB.

• Cholinergic nerve terminals uptake choline by a either a high affinity or

a low affinity choline transport system.

• The high affinity system is sodium-dependent and carrier mediated and

is unique to cholinergic cells. It is inhibited when the cell is
depolarized.

• Low affinity choline transport operate by passive diffusion; provides

many non-neuronal cells with the choline they require to carry out
processes such as the synthesis of various choline-containing
phospholipids
 ACh synthesis

• Factors regulating ACh synthesis:

• negative feedback: ACh binds to ChAT and inhibits its catalytic ability.

• Law of mass action: ACh concentration is proportional to the availability of

Acetyl CoA, choline and choline acetylcholietransferase.

• The most important limiting factor is the rate of choline high affinity uptake.

• Drugs inhibiting Ach synthesis

• Juglone: ChAT inhibitor

• Triethylcholine (TEC): Inhibits high affinity choline uptake

• The latter is more effective, reflecting the major function of choline uptake in
synthesis
 Termination of ACh activity

-Upon release, acetylcholine is hydrolyzed into choline and acetate by

acetylcholinesterase (AChE), and, to a much lesser extent, by other
nonspecific cholinesterases.
• AChE has an anionic subsite which attracts the positive charge of ACh and
an estertic site which possess the serine residue that breaks the ester bond.
• Extremely efficient enzyme, hydrolyzes 5000 molecules of ACh per
molecule of enzyme per one second.
• BUT expresses “excess substrate inhibition”-inhibited by high concentration
of ACh
 Irreversible acetylcholinesterase
inhibitors =“nerve gas”
Autonomic Central
Neuromuscular Nervous Nervous
Effects System System
Effects Effects

Twitching
• Reduced
• Headache
•

Weakness
• Vision Convulsions
•

Paralysis
• Small pupil
• Coma
•

Respiratory failure
• size Respiratory
•

Drooling
• arrest
Sweating
• Confusion
•

Diarrhea
• Slurred
•

Nausea
• speech
Abdominal Depression
organophosphorous compounds –which • •

pain Respiratory
form highly stable phosphorlyated complexes
•

Vomiting
• d
with AChE that persist for hours or more

Treatment
PAM: (2 pyridine aldoxime methiodide): displaces the inhibitor from the active site.
•

However ,doesn’t cross the BBB.

Atropine: doesn’t effect the inhibitor at all, but balances the effect of AChE by
•

blocking the muscarinic ACh receptors. Currently the most commonly used.
 Other pharmacological goodies
• Botulinum toxin A:
prevents ACh
releaseÆparalysis

• α-latrotoxin (black widow

spider venom): promotes
massive vesicular ACh
release

• Hemicholinium and
vesamicol: prevent
choline reuptakeÆlowers
ACh synthesis and stores
 Cholinergic pathway: CNS
Acetylcholine is widely distributed in the peripheral and central nervous
system. In the CNS, cholinergic neurons can be divided to several groups.
:

Main groups of ACh groups

and pathways in the CNS

1. Interneurons within striatal

complex; regulated by
glutamate and dopamine

2. Basal forebrain (including

nucleus basalis, medial
septal nucleus, substantia
inominata); source of
projection neurons to
limbic system and cortex
 Cholinergic pathway: PNS
Motor systems
Cholinergic neurons are found in the spinal ventral horn and
in cranial nerves 3-7 and 12. Thus, ACh is secreted in all
neuromuscular junctions and stimulates muscle
contraction.

Autonomic nervous system

The principle transmitter by the parasympathetic and
sympathetic pregangalionic fibers.

Released by most of the parasympathetic post ganglionic

innervating glands and smooth muscles which will be
excited or inhibited depending on the type of receptor they
present.
 Cholinergic receptors
Cholinergic receptors can be divided into 2
major categories. Muscarinic receptors
originally were distinguished from
nicotinic receptors by the selectivity of
the agonists muscarine and nicotine
respectively.
 Basic differences between
muscarinic and nicotinic receptors
Agonist Muscarine Nicotine

Response time Slow (100-250 ms) Fast (10<ms)

Receptor type Metabotropic Ionotropic

Receptor subtype M1, M3, M5 =Gq coupled 9 subtypes

M2, M4=Gi coupled

Excitatory/inhibitory Mixed Excitatory

Typical antagonists Atropine, Scopolamine Tubocurarine

distribution 9Parasympathetically 9All striated muscles

innervated cardiac and smooth
muscles (heart, iris, stomach,
bronchitis bladder etc).
9Salivary, tear and sweat
glands
9CNS
9Postganalionic
parasympathetic and
sympathetic neurons
9CNS
 Nicotinic receptors antagonists:
Curare

Curare was used as an arrow poison by indigenous forest peoples of

South America to paralyze their prey.
The main active ingredient was tubocurarine-antagonists at the nicotinic receptor.
Nicotinic ACh Receptor

-alpha forms ACh and nicotine

binding site
-cation channel
-rapidly desensitizes upon
agonist stimulation
 Nicotinic Acetylcholine Receptors
Receptor Skeletal muscle Autonomic CNS CNS
ganglion

Subunits α1,β1,δ,γ(ε) α3,α5,α7,β2,β4 α3,α4,β2,β4 α7,α8,α9

Antagonists α-Bungarotoxin Hexamethonium Dihydro-β- α-Bungarotoxin

erythroidine Mecamylamine

Mecamylamine

Agonists Epibatidine Epibatidine Epibatidine

ABT-418
 Muscarinic receptors
Muscarinic Acetylcholine Receptors

M1 M2 M3 M4 M5

Distribution Cortex, Heart Exocrine Neostriatum Substantia

hippocampus glands, GI nigra
tract

Selective Pirenzepine AF-DX 116e pF-HHSiD

Antagonists
Selective CDD-0097
Agonist Xanomelin
G protein Gαq/11 Gi/o Gαq/11 Gi/o Gαq/11

Intracellular PLCβ Adenylyl PLCβ Adenylyl PLCβ

response cyclase cyclase
inhibition inhibition
 vesamicol

botulinum-A
α-latrotoxin Nerve gas
PAM

Muscarine
Nicotine
Atropine
Curare
Scopolamine
Mecamylamine
 AADC

H1 antagonists: “anti-histamines”Ædrowsiness; H2 antagnoists=anti-nausea;

H3 antagonistsÆincrease wakefulness