Glutamate Receptors

Glutamate Receptors:

An unusual Glutamate-activated Cl channel

(parasitic helminths): Cys-loop receptor
Glutamate-activated cation channels:
Glutamate as a neurotransmitter
Glutamate receptor structure: different from
Cys-loop receptors
3 Classes of glutamate receptors:
pharmacology & physiology

Glutamate-activated Cl channels:
a target of anti-helminthic drugs
Roundworms (nematodes) and flatworms are responsible for
parasitic infections, that usually involve complex parasite
lifecycle with multiple hosts
Onchocerciasis (causing river blindness), lymphatic filariasis
(causing elephantitis), and canine heartworm are caused by
filarial nematode infections transmitted by black flies or
mosquitoes
World Health Organization Initiatives have targeted these
conditions, resulting in improvement in worldwide health
Antihelminthic drugs act by preventing muscle contraction in
the parasite larvae or altering metabolism (e.g. interfering with
microtubule function)
Another approach is antibiotics to kill symbiotic Wolbachia
bacteria, rendering the female worms sterile

Approaches to inhibit worm larvae:

Unlike vertebrates that have only
excitatory synapses at the NMJ,
roundworms have both excitatory and
inhibitory (GABA-activated Cl channels)
synapses onto skeletal muscle

Worm Glutamateactivated Cl channel

Also, Glutamate-activated-Cl channels

inhibit muscle in pharnyx directly, and
inhibit skeletal muscle via inhibitory
interneuron
Glutamate-activated Cl channels are
Cys-loop receptors, like nAChR, GABAA
R & Glycine R
Ivermectin is a Glutamate-Cl receptor
allosteric agonist used to kill microfilariae
that cause river blindness, and to lesser
extent to treat elephantitis
Piperazine, a GABA agonist at
invertebrate NMJ, also is effective
Other therapeutics include inhibitors of
microtubule function or arachidonic acid
metabolism (see126C next quarter)

Hibbs & Gouaux 2011

Glutamate
The major excitatory neurotransmitter in mammalian brain
A family of 3 types of ligand-gated cation channels;
permeable to Na+, K+. NMDA subtypes also permeable to
Ca2+
What does glutamate do?
Involved in learning & memory, and long-term potentiation, a
cellular correlate of learning
Buttoo much glutamate causes neural toxicity.
What happens if inhibit glutamate action?
Depress excitation in brain, anesthesia, but has psychotropic
actions
Can glutamate pathways be targeted to enhance
learning/memory, control Alzheimers disease, or inhibit effects
of stroke/brain trauma?

2 Families of Structurally Different Ionotropic Receptors :

Nicotinic ACh R
5-HT3 R
GABAA R
Glycine R
(& Glu-Cl R: worms)

Glutamate R
(mammalian)

5 subunits: N-ACh R, 5-HT3 R,

GABA R, Glycine R, (&
invertebrate Glutamate R)
4 subunits: Glutamate R
(mammalian)

EAAT

How is glutamate action terminated?

Does this require energy?
EAAT (Excitatory Amino Acid Transporter co-transports Na+ and glutamate

Glutamate Receptor Structure

(mammalian):
Ligand binding site:
N terminus & M3-M4 linker
Pore region:
Re-entrant loop M2
& M3
Channel is a tetramer of
homologous subunits

Pore region (shaped like an

upside-down KV channel):
Re-entrant loop M2 forms the
inner part of the pore
M3 lines the outer part of the
pore and forms the gate for
channel opening

Structure of AMPA Glutamate R

4 subunits
Structure is a dimer of dimers
Amino Terminal Domain:
involved in receptor assembly,
trafficking and modulation

Critical site in the pore:

NMDA R have an N
(asparagine) at this site. This
makes a channel that IS Calcium
permeable, blocked by Mg2+

Ligand Binding Domain:

4 binding sites for
agonist/antagonist
one within each subunit
clamshell
Transmembrane domain
Forms the pore

Sobolevsky, Rosconi, Gouaux (2009) Nature 462, 745

NMDA Receptor: Binding Sites

3 Classes of Glutamate Receptors:

Kainate R

AMPA R

A combination of 4 homologous subunits forms each

glutamate receptor.

AMPA R undergo mRNA editing.

Their mRNA encodes a Q
(glutamine). On the GluR2
subunit, this site is edited to
encode R (arginine). QR
editing introduces a positive
charge that makes channels that
are NOT Calcium permeable, not
blocked by Mg2+

NMDA R

Newer Nomenclature:

3 Types of Glutamate Receptors:

NMDA
AMPA
Kainate
NMDA Receptor

Non-Competitive NMDA Antagonists

NMDA Receptor

NMDA receptors
require glycine as
a co-agonist

Non-competitive antagonists:
Ketamine is a veterinary
anesthetic, also used for
children; drug of abuse; currently
being evaluated for treatment of
depression
Phencyclidine (PCP) is a drug of
abuse
Both are dissociative anesthetics

AMPA Receptor

Kainate Receptor

NMDA Receptor:
Glutamate binds to 2 of the subunits (GluN2) and glycine binds to the other two
subunits (GluN1 or GluN3)
Physiological background [glycine] often sufficient to serve as the co-agonist

Synaptic Physiology
Most central excitatory synapses
have both AMPA and NMDA
receptors.
AMPA R
Fast excitatory response

NMDA R
Slightly slower excitatory response
Excitatory action:
Both NMDA and AMPA produce
inward Na currents, which will
depolarize the cell membrane.

How do non-competitive
antagonists inhibit the NMDA R?
How can we learn how they
work?

Mg2+ blocks NMDA Receptor at Negative Potentials

NMDA receptors serve as

a coincidence detector:
They are only open when
the post-synaptic cell is:
a) depolarized and
b) glutamate is present

NMDA receptors cause

calcium influx, which is
important for learning &
memory, but also cause
neuronal death
At negative membrane
potentials, the electrical
driving force drives Mg2+ into
the pore and blocks it

At more positive voltages, Mg2+

is expelled from the channel,
and ions can flow through the
pore

Summary
Glutamate: inhibitory for worm muscle; allosteric agonist for
treatment of filarial infection (e.g. river blindness)
Glutamate: major excitatory neurotransmitter in mammalian brain
Structure: tetramer, cation channel, sites within pore loop
determine Ca2+ permeability & Mg2+ block
3 classes of mammalian Glutamate Receptors:
NMDA R (agonist: glutamate, aspartate, NMDA; co-agonist: glycine;
antagonist AP-5; channel blocker: PCP, ketamine), slow EPSP,
excitotoxicity, Na+, K+ & highly Ca2+ permeable, blocked by Mg2+,
widely distributed
AMPA R (agonist: glutamate, AMPA, Quisqualate; Antagonist:
NBQX, CNQX) fast EPSP, Na+ & K+ permeable but low Ca2+
permeability, widely distributed
Kainate R (agonist: glutatmate, kainate) Fast EPSP, low Ca2+
permeability, limited distribution

Use-dependent block: ketamine & PCP must gain access to

the open pore to block the channel

Calcium

Excitotoxicity