Glutamate and Anaesthesia Flashcards

1
Q

How can we see the function of glutamate receptors in cell lines?

A

Typically you would transfect cells which do not express these receptors
And apply pulse of agonists
Use whole cell voltage clamp recordings of the cell

Can be done for AMPA, NMDAR and kainite too

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Why does Mg2+ block the NMDAR?

A

Positive charge attracted to the negative membrane potential

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

What do NMDARs conduct?

A

Ca2+

K+ in the opposite direction

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What can blockage of NMDAR cause?

A

Psychotic state

ie with ket and PCP

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

In addition to glutamate, what else does the NMDAR require to stay open?

A

A coagonist - glycine or D-serine

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

How do we know that NMDAR activation requires agonist and coagonist binding?

A

From whole cell voltage clamp recordings

Noted that in the absence of glycine, glutamate current is not persistent as it does not activate NMDARs

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Where does glutamate bind to the NMDAR?

A

GluN2 subunit

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

Where does glycine bind to NMDAR?

A

GluN1 subunit

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

How do we know that Mg2+ blocks slow NMDAR transmission?

A

Measurement of NMDAR EPSCs in neurons
Cells in solutions containing Mg2+ only demonstrate a fast EPSC which is dampened by a AMPA antagonist (CNQX)
In Mg2+ free solution the EPSCs have a slow and a fast component
The CNQX here has no affect on the slow component

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Which part of the glutamate receptor is the part which makes up the ion channel?

A

TM2

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What are most GluA2 subunits edited to in an adult?

A

GluAR

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

GluAR is calcium impermeable

T/F

A

True

GluA1,3,4 and unedited GluA2(Q) are calcium permeable

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

What is the edit made in GluA2?

A

Adenosine to arginine in the TM2 re-entrant loop

Q607R

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Why don’t polyvalent cations (such as spermine) block GluA2R?

A

It is positive so is repelled by the positive charge of arginine

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

What do AMPARs mediate?

A

Fast synaptic transmission

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

What is the difference between the function of edited and non-edited AMPA subunit GluA2?

A

Q - calcium permeable and blocked by internal polyamines

R - calcium impermeable, not blocked by internal polyamine and reduced single channel conductance

17
Q

What shape of structure do the S1 and S2 domains take on the AMPAR?

A

Clamshell structure which shuts when bound

18
Q

Which subunits are most NMDARs made from?

A

GluN1 and GluN2

19
Q

What is there in the NMDAR equivalent of Q/R site?

A

Asparagine (N)

site of Mg block

20
Q

What does IV ketamine induce?

A

A dissociative state with marked sensory and memory loss

Without complete LOC

21
Q

Describe the effects of ketamine on GABAa receptors?

A

(Little to) none

22
Q

How does ketamine affect NMDAR?

A

Voltage dependent block

23
Q

What is the state of the NMDA channel when ketamine applied?

A

Equilibrium between bound and open (ar) and open but blocked (2ard)

24
Q

What “anaesthetic” competes with glycine for the co-agonist site on the GluN1 subunit?

A

Xenon

25
Q

What is normally used for the induction of anaesthesia and why?

A

IV drugs
Propofol, etomidate, thiopental
They produce a state of unconsciousness within 20 seconds

26
Q

What is used for maintenance anaesthesia?

A

Inhalational agents
Like isoflurane or sevoflurane

Or continuous propofol

27
Q

What is the main difference between nitrous oxide and isoflurane?

A

Isoflurane acts on GABA

Nitrous oxide doesnt

28
Q

Outline AMPAR trafficking in and out of the postsynaptic density

A

Newly made recs transported in vesicle by molecular motors on microtubules

Exocytoses into dendritic shaft (then moves randomly at cell surface)

Stabilised by diffusion trapping via interactions with scaffold proteins

Diffusion receptors internalised at the extrasynaptic endocytic zones by clarithirin dependent endocytosis

These are then recycled back by exocytosis or tagged for breakdown

29
Q

What may cause AMPAR trafficking?

A

Neuronal activity
Stress hormones
Neurodegen disease
Drugs

30
Q

Which drugs may cause AMPAR trafficking?

A

Ketamine
Cocaine
Tianeptine

31
Q

How can LTP be induced?

A

High frequency theta burst stimulation of the Schaffer collateral pathway from CA3

32
Q

What induces LTD?

A

Repetitive LFS

33
Q

Which is associated with an increase in synapse number?

LTP or LTD?

A

LTP

34
Q

What occurs with stabilising AMPARs in the synapse?

A

Interactions of C terminus and various interactor proteins

35
Q

Which ampakine may increase QoL in HD?

A

CX929
Improves LTP in mutant huntingin and improves cognition and performance in NOR in CAG140 mice

(Nekooki-Machida 2009)

36
Q

Ketamine acts on NMDAR, the metabolite HNK acts on…

A

AMPAR

37
Q

How do we know this is the receptor where HNK works?

A

Administration of ketamine and metabolite and monitored the EPSCs of the different receptors