Neuroscience essay title:

1. Describe the structures, mechanisms and function of neurotransmission at

a glutamatergic synapse.

Glutamate serves to be one of the primary excitatory neurotransmitter in

the brain and spinal cord. It is vastly abundant in about 70% of cortical
neurons, which signifies its importance in mediating important brain
functions such as cognition, particularly memory, coordination, sensory
transmission and emotion. It is vital to comprehend the processes
underlying a glutamatergic synapse from synthesis to degradation; its
dysfunction could be detrimental and therapeutics to counter this could
be particularly beneficial. Moreover, excess accumulation of extracellular
glutamate is toxic. This phenomenon is present in some significant
neurodegenerative diseases, so understanding this mechanism is crucial.
Glutamate, a non-essential amino acid, cannot cross the blood-brain
barrier, so any production of glutamate must take place from precursors
within the brain. Glutamate can be synthesised from two primary sources.
The first source relies on the glucose metabolism within the brain. This is a
reliable source because of the constant activity of aerobic respiration to
produce ATP in the brain. The precursor arises from the TCA cycle as an
intermediate: a-ketoglutarate. A-ketoglutarate is transaminated into
glutamate in the mitochondria of the nerve terminals through the enzyme
GABA-transaminase. is an intermediate of the TCA cycle: a-oxoglutarate.
The second process of synthesising glutamate involves the conversion of
glutamine to glutamate in the astrocytes. This is a recycling process,
because glutamate is taken up by the astrocytes, and is converted to
glutamine by the glial-specific enzyme glutamine synthetase. Glutamine
diffuses out into the neurons, where it is converted back to glutamate by
glutaminase. This method of synthesis is not completely stoichiometric
because it is mere recycling, so the quantity of glutamate lost by glial
cells cannot be replenished.
Once glutamate is synthesised, it is essential to store it so that its
extracellular quantities are tightly regulated to avoid any unnecessary
disturbances in brain function. Glutamate is actively taken up into vesicles
in the presynaptic nerve terminals through the transporters vGlut1-vGlut3.
These secondary active transporters use the electrochemical gradient of
protons to transport glutamate against its concentration gradient. The
maintenance of the proton gradient is mediated by the H+ pump, which
uses ATP to pump protons into the vesicle so that protons can diffuse out
down its gradient and glutamate can enter through one of the Vglut
transporters. Furthermore, since glutamate is negatively charged, the high
positive potential within the vesicles would potentiate the accumulation of
glutamate inside. There are about 1000 glutamate molecules present in
each vesicle, which highlights the high concentration of this
neurotransmitter in vesicles at the nerve terminal.

The release of these vesicles are now dependent on the stimulation of the
nerve terminals. When an action potential arrives, the membrane
depolarisation subsequently opens voltage-gated calcium channels to
enable the entry of calcium ions. These ions stimulate the migration of the
vesicles to the presynaptic membrane. Here, the vesicles are anchored by
SNARE proteins, and glutamate is released from the vesicles through
exocytosis onto the synaptic cleft.
Glutamate diffuses across the synaptic cleft and binds to one of the
multiple post-synaptic receptors. This propagates an action potential in
the post synaptic terminal, provided that the signal crosses the threshold
of the post-synaptic neuron. Glutamate is then removed from the synaptic
cleft to terminate the excitatory signal. Glutamate is taken up primarily by
the astrocytes through excitatory amino acid transporters (EEAT1 and
EEAT2). Here, the glutamate is converted into glutamine in the recycling
process described above. Glutamate is also taken up into nerve terminals
through EEAT3 and EEAT4, although this is less significant. EEAT
transporters function by co-transporting Na+ and H+ ions with the
glutamate, and counter-transporting K+ out of the cells. Finally, glutamate
can diffuse away to act in other synapses.
The receptors that glutamate bind to in the post-synaptic membrane have
unique properties that complement their role in excitatory transmission.
These receptors can be either ionotropic (ion channels) or metabotropic
(G-protein coupled receptors). Ionotrophic are characteristic for fast
transmission, and involve the agonist binding to the channel, which would
open the channel. The three glutamate ionotropic channels are NMDA,
AMPA and Kainate, named after the agonist that binds to them. However,
glutamate is a transmitter common to all 3 receptors. Each of these
receptors are tetrameric; they have 4 subunits, with each subunit have 3
transmembrane spanning domains.
AMPA and NMDA receptors co-exist in essentially all neurons in the CNS,
because their functions are inter-linked in depolarising the post-synaptic
membrane. Kainate receptors are located pre-synaptically on GABA
secreting nerve endings and post-synaptically at various sites. Kainate
and AMPA receptors can also be found on glial cells, but NMDA receptors
are found only in neurons.
AMPA has GluA1-GluA4 subunits, and is known for its fast depolarisation
and decaying. NMDA has 2 GluN1 and 2 GluN2 subunits. Additionally,
NMDA receptor requires the binding of glycine or D-serine to GluN1
subunit for activation. The unique structural feature of NMDA also includes
the blockage of the channel by Mg2+ ion like a plug in a hole. Mg2+ is
expelled only when the membrane potential depolarises past -50mV.
Initially, when glutamate arrives at the post-synaptic membrane, it binds
to both AMPA and NMDA. AMPA channels open immediately, which allows
an influx of Na+ ions and an efflux of K+ ions, triggering the fast EPSP.
Very few Ca2+ ions can flow through AMPA receptors. When the same

glutamate binds to NMDA receptors, due to the small depolarisation at the

membrane, the Mg2+ block is present and the ions cannot flow through.
AMPA is rapidly decaying, so frequent stimulation increases the membrane
depolarisation until it reaches the threshold of expelling Mg2+ out of
NMDA. NMDAs slow activation and decaying enable this phenomenon of
summation to occur. Once the channel is completely open, there is a large
depolarisation due to large influxes of Na2+ and Ca2+. Ca2+ plays a vital
role intracellularly as it triggers a phosphorylation cascade to activate
various kinases. It is very important to ensure that NMDA channels have
these protective barriers in stabilising its inactive state, because excess
influxes of Ca2+ can damage the neurons in detrimental ways.
Kainate receptors are also tetromeric with homomeric or heteromeric
GluK1-GluK5 subunits. The roles of kainate receptors are less well
understood in the CNS, but it is believed that it is involved in the
modulation of neurotransmitters of excitatory and inhibitory synapses in
the pre-synaptic terminal, and the excitation of post-synaptic neurons
through ambiguous G-protein coupled effects.
Metabotrophic 3 types. Group 1 Gq G-protein, Ip3-Dag excitatory
Group 2 Gi G-protein, decrease adneylate cyclase
Transmembrane, no subunits. MGlu1-8.
Function: presynaptically function as inhibitory autoreceptors.
Postsynaptically, function variably according to cell type can inhibit K+
channels which increase excitability, or vice versa.
They are also said to be involved in the production of synaptic plasticity,
the concept of increasing or decreasing the strength of a synapse over
time. This is promiment in the hippocampus and cerebellum. Defects in
Mglu1 can lead to deficits in spatial learning and severe motor
incoordination.
Long term potentiation is a concept * Prolonged activation of AMPA
receptors leads to strengthening of the synapse via altered gene
transcription and structural changes to the synapse. Sustained activation
of AMPA receptors can influence the activation of calmodulin kinases,
along with GTPases, to play a part in actin cytoskeleton and gene
expression. It also increases the conductance and number of AMPA
receptors, so that a small level of glutamate can trigger a larger
depolarization. Underlies the mechanism of long-term memory and
learning in hippocampus.
* Relates to Ca2+ entry into the cell
* If too much glutamate is around, NMDA receptors are activated and
Ca2+ enters the cell
* Too much intracellular Ca2+ leads to activation of Ca2+-dependent
apoptosis pathways (e.g. activation of caspases)

* Occasions where there is too much glutamate include stroke, where

inability of partially ischemic cells to maintain Na+ electrochemical
gradient. So cannot remove glutamate from extracellular fluid. So
glutamate accumulates excitotoxicity and cell death in penumbra.
Excessive glutamate receptor activation can also contribute to
pathophysiology of neurodegenerative diseases ALS, PD, alzhemiers.
The glutamatergic synapse, apart from holding great relevance in causing
diseases, can also be used in the development of therapeutics. Riluzole
antagonizes NMDA receptors through its voltage gated channel blocking,
and can slow progression and improve life expectancy of ALS patients.
Memantine is another NMDA receptor antagonist used to slow down
progressive decline of alzhemiers. Amantadine has been used as an
adjunct with Levdopa to slow down PD.
Many clinical trials to explore these receptors further, but so far they have
been disappointing. Recently, a novel binding mode has revealed 2
subclasses of NMDAreceptors antagonists, which can be further analysed
to explore future therapeutics ((Stroebel et al., 2016).