Presynaptic terminal

Glial

Postsynaptic neuron

GABA and GABA receptors

Lecture 1. GABAA receptors Lecture 2. GABAB receptors Lecture 3. GABA homeostasis Lecture 4. Modulation of GABAergic synaptic transmission

What is GABA?

-aminobutyric acid Inhibitory neurotransmitter. ~1/3 of synaptic transmission in the brain is mediated by GABA. Neurons that synthesize and release GABA is called GABAergic neurons.

GABA receptors
GABAA receptors
Ligand-gated ion channels Fast synaptic inhibition

GABAB receptors
GTP-binding protein coupled receptors Slow synaptic inhibition

GABAC receptors
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Lecture 1. GABAA receptors

molecular structures location and function single channel recording pharmacology two types of inhibition

Benzodiazepines Barbiturates Neurosteroids

Anesthetics

GABAAR is ligand-gated ion channel

Receptor Transmitter Pore Channel

Extracellular side

Gate

Cytoplasmic side

Molecular structure of GABAA receptors

1-3

Moss & Smart 2001

 subunits are part of GABAA receptor family

1-3
Moss & Smart 2001

structure function pharmacology

Dendrogram of the deduced amino acid sequences of GABAAR subunits. from Cherubini and Conti 2001

Distribution of GABAAR -subunit mRNA in rat brain

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Molecular structure of GABAA receptors

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.
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Location
Presynaptic terminal Glial Synaptic cleft

GABAAR IPSP

Cl

Postsynaptic neuron

Cl- and HCO312

Nernst equation
Cl 10 mM
-

Cl125 mM

RT ln [Cl]o zF [Cl]i [Cl] o ECl = -60 log [Cl] i ECl = -66 mV ECl =

z = charge of diffusible ion (Cl- = -1) R = universal gas constant T = absolute temperature F = Faraday's Constant
13

Current passing

recording

Current passing

recording

Inhibitory interneuron

Motor neuron

IPSP AP

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Reversal potential of IPSP

Postsynaptic potential -35 -55 -74 -99 (mV) Current clamp Voltage clamp
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Postsynaptic current

Cl flux

ECl-

IPSP reduces cell excitability

Membrane hyperpolarization
drive membrane potential away from the threshold potential.
Threshold potential

Reduction in membrane resistance

reduce the excitatory input. This is known as shunting inhibition.

ECl-

Threshold potential

ECl16

Single channel recording

Open GABA Closed

Closed

Bound

Open

Bound

Open

Bound

Closed
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ligand binding sites on GABAA receptors

benzodiazepine

picrotoxin

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GABAA receptor pharmacology

Response (%)
100

Maximum Response

Agonists
GABA, muscimol

50

Antagonists
Response (%)

Bicuculline, picrotoxin, gabazine

10

KD

1000

Dose

100

Modulators
Zn2+ Neurosteroids Benzodiazepines Anesthetics Barbituates Alcohol

affinity
50

affinity
1 10 100 1000

Dose

Response (%)

100

efficacy efficacy
1 10 100 1000

50

Dose

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Phasic and tonic inhibitions

Bicuculline
20 pA 40 ms
1 min 50 pA

high agonist dose ~1 mM quantal release

Action potentialdependent IPSPs Action potentialindependent mIPSPs

low agonist dose ~1M unknown mechanisms

reverse uptake spill over extracellular matrix channel spontaneous open

synaptic receptors sensitive to gabazine

extrasynaptic receptors insensitive to gabazine

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GABAAR and disease

GABAAR is a major target for developing therapeutics.
pain epilepsy anxiety depression sleeping disorders

Mutations in GABAARs are found to be linked to epilepsy.

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GABAAR 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 GABAA receptor subunit correlate with certain type of epilepsy.
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Questions

23

Lecture 2. GABAB receptors

molecular structures location and function modulation

24

GABABRs are G-protein coupled receptors

Transmitter Receptor G protein
Channel

GTP NH2
P P P

Gate

Extracellular side

Cytoplasmic side COOH

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Molecular structure of GABAB receptors

Heterodimer linked by coiled-coil domain GABAB1a-f GABAB2, 35% homology with GABAB1 coupled to Gi/Go

Marshall, FH et al, 1999 26

Function of GABABRs
Ca GABABR
2+

GTP P P
P

Adenylyl cyclase
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GABABRs - postsynaptic
GABABR

fast IPSP ECl EK

slow IPSP
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GABABRs - presynaptic
GABAergic
GABAARs

Glutamatergic GluRs

GABABR IPSP

GABABR
Threshold

EPSP

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GABABR function
opening K+ channels in the postsynaptic membrane. closing Ca2+ channels in the presynaptic terminal.
GABAergic: autoreceptor glutamatergic: heteroreceptor

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GABABR pharmacology
Agonists
GABA, (-)baclofen, APPA

Antagonists
saclofen, phaclofen, CGP35348, CGP55845A

Modulators
CGP7930

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GABABR and disease

GABABR agonist
antispasticity antinociceptive suppression of drug craving

GABABR antagonist
suppress absence seizure in animal models

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GABABR Summary
G-protein coupled receptors. Heterodimer with GABAB1 and GABAB2. Mediate slow IPSP via opening K+ channel at postsynaptic membrane. Decrease synaptic release via inhibit Ca2+ channels in the presynaptic terminal. targets for therapeutic agents.

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Function of GABAARs and GABABRs

Under several conditions GABARmediated response can be excitatory.
GABAARs GABABRs
EIPSC higher than the threshold (Cl- or HCO3-). Disinhibition Presynaptic inhibition on inhibitory neurons Activating K+ channel may recruit T-type Ca2+ channel to induce oscillation in thalamus. Disinhibition
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Reference books
Principles of neuroscience
4th Edition Eric R. Kandel Jame H. Schwartz Thomas M. Jessell New York: Elsevier Sunderland: Sinauer Associates
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From neuron to brain

4th Edition John G. Nicholls Robert Martin Bruce G. Wallace Paul A. Fuchs