Imaging Localisation

GluR1-4 GluR2

5 m

Surface and total distribution of AMPA receptors in cultured hippocampal Neurones. Noel et al.1999, Neuron 23 p.365-376

EGFP-Sindbis virus infection, CA1 neuron hippocampusA. Terashima, A. Griffiths & F. Duprat, 2000

7 days long term culture. Injection in CA3 pyramidal neuronsCourtesy of Akira Terashima, 1999

ElectrophysiologyFunctional properties

Long Term Potentiation: LTP

Long Term Depression: LTD

Frequency (Hz)

Cha

nge

in E

PS

Psl

ope

(%)

+30

0

-30

100

E

PS

C A

mpl

itud

e (%

)

Time (min)-20 0 20 6040

Time (min)-20 0 20 60

100

E

PS

C A

mp

litu

de

(%)

40

From Bear 1996 Proc. Natl. Acad. Sci. USA 93, p.13455

Synaptic plasticity

Spontaneous activities

mEPSP, mEPSC, mIPSP, mIPSC

sEPSP, sEPSC, sIPSP, sIPSC

1) Na+-channel independent: minis= spontaneous neurotransmitter release (TTX)

Evoked activity

EPSP, EPSC, IPSP, IPSC

= activity of N synapses

Time (min)

EP

SC

am

pli

tud

e (p

A)

10 20

-80

-20

EPSC amplitude (pA)

Nor

mal

ised

fre

qu

ency

5

15

25

20 ms

5 pA

-80 -40 0

Depression of EPSCs by pep2mFrom Lüthi et al.1999 Neuron 24, p. 392

2) Na+-channel dependent:spontaneous= spontaneous action potential (no TTX)

Inter-mEPSC Interval (s) mEPSC Amplitude (pA)Cu

mu

lati

ve p

rob

abil

ity

From Noel et al. Neuron 23, p. 2

MRC Centre for Synaptic Plasticity,University of Bristol, England

Dr Fabrice DUPRAT, March 2000

x 4 =(x 5 ?)

GluRx

Molecular properties

Molecular Biology / Biochemistry

IonotropicGlutamate Receptors

MetabotropicGlutamate ReceptorsGABA Receptors

GABAA GABAB

Non NMDA

AMPA Kainate I II III

GluR1,2,3,4

Edited GluR2

GluR5,6,7

KA1,2

mGlu1,5 mGlu2,3 mGlu4,6,7,8

Ca2+ permeable

Ca2+ impermeable

Low affinity, Ca2+ permeable

High affinity

Na+, K+

(Ca2+, Mg2+)Na+, K+

(Ca2+)Gq/11

PLCGi/o

ACGi/o

AC

Su

bu

nit

sS

ign

als

Ago

nis

tsA

nta

gon

ists

Competitive Modulatory

NMDA,L-Glutamate,L-Aspartate,

Ibotenate

Glycine,Serine

D-AAD-AP5,

CGS19755,CGP37849,CGP39551,AP7, CPP

7-chloro kynurenate,

CNQX,NO, H+, Mg2+

GYKI53655*,GYKI52466*,

NBQX, CNQX

LY382884,LY294486,

NS102,NBQX,CNQX

Quisqualate,(S)-DHPG,Glutamate,

ACPD, Ibotenate,

CHPG5

AIDA,LY3673851,

MPEP5*,CPCCOEt1*

MCPG

LY379268,L-CCG-1,DCG-IV,

(2R,4R)-APDC,LY354740 ,Glutamate,

(1S,3S)-ACPD

MCCG,LY341495,PCCG-IV

MCPG

L-AP4 ,L-SOP,

Glutamate,L-CCG-1

LY341495,MAP4,CPPG

NR1 a,b,c,d,e,NR2 A,B,C,D

Ca2+

K+, Na+

NMDA

: 6 s/u,: 4 s/u

Bicuculline,SR95531

GABAB R1a,b,c,dR2

1,2,31 1,2,3 1

GABA

GABA,Isoguvacine,

Muscimol

Flunitrazepam,Zolpidem,Abecarnil

SitesBZ

Cl-

Benzodiazepines, Flumazenil,

ZK93426

Gi/o

AC

L-Baclofen,GABA

PhaclofenCGP35348,

2-OH saclofen,CGP55845

Nam

es

Types:

Str

uct

ure

s

Quisqualate,AMPA,

Domoate,Glutamate,

Kainate

Kainate,Glutamate,Domoate,AMPA,ATPA

Edited GluR5,6

Low affinity, Ca2+ impermeable

(* non competitive, 1,5 mGlu1 or 5 only)

F. Duprat, march 2000

F. Aguila & F. Duprat, 1998

mGluRI

mGluRII

mGluRIII

mGluRII

mGluRI

mGluRI

mGluRII

mGluRIII

Na+

K+

Na+

Ca2+

K+

Cl-

Na+

Ca2+

K+NO

K+

Ca2+

K+

Cl-

Cl-

ICl V,Ca

INa T,P

ICa T,N,L,R,Q,P

IK VA,D,M,C,L,AHPIK leak,ATP,IR,Ca

IQ

GABAA

GABABGABAA

SST,NPY,adenosine,5HT,ACh

Cl-

GABAA

EAAC1EAAT4

GLASTGLT1

Glu

Glu

[Ca2+]in

Gi/oGi/o

Gi/o

Gi/o

Gi/o

Gq/11

Gq/11

Gq/11

Glu

Glu

Glu

GABA

Glu

G

ActionPotential

Nt Release

ActionPotential

[Ca2+]in NM Release

Gi/o Gq/11

+

+

++

+

spine

Postsynapticmembrane

Glial cell

Inhibitory synapse

Excitatory synapse

epsp

slow ipsp

fast ipspV

t

Vt

Na+

(Ca2+)

Ca2+

Na+

AMPA

NMDA

Kainate

PSD

(Ca2+)

+ Na+

GAT1

Glu

Na+

Kainate(Ca2+)

F. Duprat, march 2000

Synapses

Dendrite

Axon

Axon

Network of cortical neurons

Rat CA1 Pyramidal cell labeled with EGFPTwo photon laser scanning microscopyFrom Maletic-Savatic et al. 1999, Science 283, p.1924

3D representation of the torpedo nicotinic acetylcholine receptor

From Green et al. 1998, Neuron 20, p.427-444

Nervous system

Synaptic bouton in rat CA1 stratum radiatumElectron microscopy. From Shepherd et al.1998, J. Neuroscience 18, p.8302

Rat CA3 Pyramidal cell3D reconstruction of multiple spine boutonsFrom Toni et al. 1999, Nature 402, p.423

Axon

Dendrite

Presynapticbouton

Postsynapticspine

Hippocampal neuron in culture (P10-14)

Courtesy of Lisa Pickard, 1999

Blind Patch Clamp

Intracellular recordings

Patch Clamp

High resolution Patch Clamp

EPSC

50 p

A

50 ms

100 200

2

4

EP

SP

slo

pe

(m

V/m

s)

Stimulus duration (s)

100 200

1

Fib

er

volle

y (m

V)

Stimulus duration (s)

2

100 200

4

8

12

Po

pu

latio

n s

pik

e (

mV

)

Stimulus duration (s)

Extracellular recording

Field recording

Adapted from E.J. Coan,PhD thesis 1988, UOB

Somatic extracellular recording

Dendritic extracellular recording

mV

msf.v.

Adapted from C.H. Davies, PhD thesis 1991, UOB

4 m

V

10 ms

fEPSP

Subthreshold Suprathreshold

mV

msp.s.

2 m

V

10 ms

fEPSP

Subthreshold Suprathreshold

Neurons in culture

Preparations

Thalamocortical slices

Hippocampal slices

Adapted from Agmon & Connors 1991,Neurosci. 41, pages 365-379

From G. Paxinos & C. Watson,The Rat Brain, 1998

55°

10°

From Feldmeyer et al. 1999, J. Physiol. 521, page 171

Cytochrome C oxidase stainof a fixed rat barrel cortex

From Agmon & Connors 1991,Neurosci. 41, page 370

Stimulation

Patch clamp Field recording

CA1CA1CA2CA2

CA3CA3

DGDG

Acute brain slices

Overnight slice culture

Long term slice culture

NeuronalNetwork

Neuron

Dendritictree

Receptors

Synapse

DNA

Brain

From H. ELLIS, B. LOGAN & A. DIXON “Human cross-sectional anatomy" 1991, Ed. Butterworth Heinemann

Spine

Glial cell

Dendrite

Axon

PSD

VesiclesMitochondria

Varicosity

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