1

1
Presynaptic terminal
Glial
Postsynaptic neuron
2
GABA and GABA receptors
Lecture 1. GABA
A
receptors
Lecture 2. GABA
B
receptors
Lecture 3. GABA homeostasis
Lecture 4. Modulation of GABAergic
synaptic transmission
2
3
What is GABA?
Inhibitory neurotransmitter.
~1/3 of synaptic transmission in the brain is mediated by
GABA.
Neurons that synthesize and release GABA is called
GABAergic neurons.
-aminobutyric acid
4
GABA receptors
GABA
A
receptors
Ligand-gated ion channels
Fast synaptic inhibition
GABA
B
receptors
GTP-binding protein coupled receptors
Slow synaptic inhibition
GABA
C
receptors
3
5
molecular structures
location and function
single channel recording
pharmacology
two types of inhibition
Lecture 1. GABA
A
receptors
6
Benzodiazepines
Barbiturates
Neurosteroids
Anesthetics
4
7
GABA
A
R is ligand-gated ion channel
Cytoplasmic side
Extracellular side
Channel
Pore
Receptor
Transmitter
Gate
8
Molecular structure of GABA
A
receptors
1-3
Moss & Smart
2001
5
9
subunits are part of GABA
A
receptor family
1-3
Moss & Smart
2001
Dendrogram of the deduced amino
acid sequences of GABA
A
R subunits.
from Cherubini and Conti 2001
structure
function
pharmacology
10
Distribution of GABA
A
R -subunit
mRNA in rat brain
6
11
Multiple subunits: 16, 13, 13, , , , ,
13
Each subunit contains 4 putative
transmembrane domains, TM2 is believed to
form the lining of the channel.
Hetero- or homo-oligomeric proteins.
Pentamer with :: at a ratio of 2:2:1
subunit composition determines functional
properties and pharmacology.
Molecular structure of GABA
A
receptors
12
Location
Cl
-
Presynaptic terminal
Glial
GABA R
A
Postsynaptic neuron
Synaptic cleft
IPSP
Cl
-
and HCO3
-
7
13
Nernst equation
Cl
10 mM
-
Cl
125 mM
-
E =
Cl
RT [Cl]
o
zF [Cl]
i
ln
E =
Cl
[Cl] o
[Cl] i
log
-60
E =
Cl
-66 mV
z = charge of diffusible ion (Cl
-
= -1)
R = universal gas constant
T = absolute temperature
F = Faraday's Constant
14
recording
recording
Inhibitory
interneuron Motor
neuron
Current
passing
Current
passing
AP
IPSP
8
15
Reversal potential of IPSP
-55
-35
-74
-99
(mV)
E -
Cl
Postsynaptic
potential
Postsynaptic
current
Cl
flux
-
Current clamp
Voltage clamp
16
IPSP reduces cell excitability
Membrane
hyperpolarization
drive membrane potential
away from the threshold
potential.
Reduction in membrane
resistance
reduce the excitatory input.
This is known as shunting
inhibition.
Threshold potential
Threshold potential
E -
Cl
E -
Cl
9
17
Single channel recording
GABA
Closed Open Bound Bound
Bound Open Closed
Open
Closed
18
ligand binding sites on GABA
A
receptors
benzodiazepine
picrotoxin
10
19
GABA
A
receptor pharmacology
Agonists
GABA, muscimol
Antagonists
Bicuculline, picrotoxin,
gabazine
Modulators
Zn
2+
Neurosteroids
Benzodiazepines
Anesthetics
Barbituates
Alcohol
10 1 1000
100
0
R
e
s
p
o
n
s
e

(
%
)
Dose
50
100
0
R
e
s
p
o
n
s
e

(
%
)
Dose
50
100
0
R
e
s
p
o
n
s
e

(
%
)
Dose
50
K
D
Maximum Response
affinity
affinity
efficacy
efficacy
10 1 1000 100
10 1 1000 100
20
Phasic and tonic inhibitions
high agonist dose ~1
mM
quantal release
Action potential-
dependent IPSPs
Action potential-
independent mIPSPs
synaptic receptors
sensitive to gabazine
low agonist dose ~1M
unknown mechanisms
reverse uptake
spill over
extracellular matrix
channel spontaneous
open
extrasynaptic receptors
insensitive to gabazine
20 pA
40 ms
Bicuculline
5
0

p
A
1 min
11
21
GABA
A
R and disease
GABA
A
R is a major target for developing
therapeutics.
pain
epilepsy
anxiety
depression
sleeping disorders
Mutations in GABA
A
Rs are found to be linked to
epilepsy.
22
GABA
A
R Summary
GABA-gated anion channels.
The primary inhibitory receptors in the
mammalian brain.
Pre-, post-synaptic and extrasynaptic area,
mediating inhibitory postsynaptic potentials
(IPSPs) and tonic inhibition.
Important targets for therapeutic agents.
Mutations in the genes encoding GABA
A
receptor subunit correlate with certain type of
epilepsy.
12
23
Questions
24
Lecture 2. GABA
B
receptors
molecular structures
location and function
modulation
13
25
GABA
B
Rs are G-protein coupled receptors
Extracellular side
Cytoplasmic side
NH2
COOH
Receptor
Transmitter
P
P
P
GTP
G protein
P
Channel
Gate

26
Molecular structure of GABA
B
receptors
Heterodimer linked
by coiled-coil
domain
GABA
B1a-f
GABA
B2
, 35%
homology with
GABA
B1
coupled to G
i
/G
o
Marshall, FH et al, 1999
14
27
Function of GABA
B
Rs
P
P
P
GTP

Ca
K
2+
+
GABA R
B
Adenylyl cyclase
28
GABA
B
Rs - postsynaptic
GABA R
B
fast IPSP
slow IPSP
E
E
Cl
-
K
+
15
29
GABA
B
Rs - presynaptic
GluRs
EPSP
Threshold
GABA R
B
GABA Rs
A
IPSP
GABAergic
Glutamatergic
GABA R
B
30
GABA
B
R function
opening K
+
channels in the postsynaptic
membrane.
closing Ca
2+
channels in the presynaptic
terminal.
GABAergic: autoreceptor
glutamatergic: heteroreceptor
16
31
GABA
B
R pharmacology
Agonists
GABA, (-)baclofen, APPA
Antagonists
saclofen, phaclofen, CGP35348, CGP55845A
Modulators
CGP7930
32
GABA
B
R and disease
GABA
B
R agonist
antispasticity
antinociceptive
suppression of drug craving
GABA
B
R antagonist
suppress absence seizure in animal models
17
33
GABA
B
R Summary
G-protein coupled receptors.
Heterodimer with GABA
B1
and GABA
B2
.
Mediate slow IPSP via opening K
+
channel at
postsynaptic membrane.
Decrease synaptic release via inhibit Ca
2+
channels in the presynaptic terminal.
targets for therapeutic agents.
34
Function of GABA
A
Rs and GABA
B
Rs
Under several conditions GABAR-
mediated response can be excitatory.
GABA
A
Rs
E
IPSC
higher than the threshold (Cl
-
or HCO
3
-
).
Disinhibition
GABA
B
Rs
Presynaptic inhibition on inhibitory neurons
Activating K
+
channel may recruit T-type Ca
2+
channel to induce oscillation in thalamus.
Disinhibition
18
35
Reference books
Principles of
neuroscience
4
th
Edition
Eric R. Kandel
Jame H. Schwartz
Thomas M. Jessell
New York: Elsevier
From neuron to
brain
4
th
Edition
John G. Nicholls
Robert Martin
Bruce G. Wallace
Paul A. Fuchs
Sunderland: Sinauer
Associates