Development of novel animal models

of glutamatergic central nervous system

disorders using in vivo siRNA and
transgenic approaches.

Sixth Framework Programme for research and technological development,

Capacities programme, Regions of knowledge.
CONTEXT
Mental disorders are an increasing medical problem but research into their neurobiology and the development of novel
drug treatments is restricted by the lack of suitable animal models.

AGLAEA is developing and characterising models of selective partial knockdown of specific components of the glutamatergic
system in mice. These models will provide better understanding of the important role of glutamate neurotransmission in
disorders such as schizophrenia, anxiety and cognitive dysfunction.

AGLAEA will provide the

pharmaceutical industry and
the academic environment
with new models suitable
for both testing novel thera-
peutic compounds and the
investigation of the neuro-
biological and neurochemical
basis of specific psychiatric
disorders.

shRNA using a viral vector

Construct 1:
Lentiviral construct which dri-
ves the rtTA gene under the
control of CaMK II ∝ promoter LTR CaMKII rtTA2 IRES mTan LTR
(Neuronal Specific Protein).
IRES (Internal Ribosome
NSP
Entry Site) and mTan reporter
are also incorporated.
loxP loxP

Construct 2:
pSico system to develop Pol III LTR Pol III CMV GFP shvGLUT LTR
driven CMV-GFP shVGLUT. The
integration of this construct Cre-Recombination Cre-Recombination
following transduction will be
monitored by green fluores-
cence. + Dox
+ LTR Pol III shvGLUT LTR
Mouse ES will be transdu-
rtTA/ TRE-Cre
ced simultaneously with the shVGLUT Tet inducible neuronal specific shRNA knock-down
two constructs and successful
transduction will be identified
by yellow fluorescence. Experimental approach adopted to produce inducible neuronal specific
shRNA knockdown.

Transgenic vectors for the development of inducible transgenic models

EXPERIMENTAL
APPROACH
The aim of the collaboration
is to produce, by use of RNA
WP1: siRNA/shRNA development
interference of the glutama- and in vitro validation
te membrane and vesicular
transporters, inducible mouse
models of hyper- and hypo- WP2: In vivo siRNA/shRNA
knockdown of glutamate WP5:
glutamatergic states. These transporters Management,
models will be validated using exploitation,
relevant behavioural, neuro- WP3: Functional characterisation dissemination
of the siRNA/shRNA-mediated
chemical and immunohisto- knockdowns
chemical tests including fur-
ther evaluation by functional WP4: Generation and
magnetic resonance imaging characterisation of transgenic mice
(fMRI).

1 Fabrication of
the glutamate
microsensor.

Functional
2
Magnetic
Resonance
Imaging 1 2
System (fMRI)

PROGRESS REPORT
• In vitro studies have identified suitable siRNA and shRNA
constructs for the glutamate membrane transporter (EAAT2/
GLT-1) and the vesicular membrane transporter (VGLuT1)
respectively.
• Behavioural studies indicate that disruption of EAAT2 re-
sults in an anxiogenic behavioural profile while shRNA direc-
ted at the VGLuT1 impairs specific aspects of cognition.
• Further validation of the two models is in progress together
with functional characterisation of the mechanisms involved
in the behavioural effects.
• The final aim is to produce an inducible transgenic mouse
showing partial loss of VGLuT1 expression as a model of the
cognitive deficits observed in schizophrenia.
CONSORTIUM
The AGLAEA consortium
brings together three
high tech SMEs:
FR (psychopharmacology
and behaviour),
HU (transgenic mice),
NL (neurochemistry) and
two academic groups:
NL (neurochemistry),
UK (molecular biology,
neuropharmacology
and behaviour) together
with management
provided by ALMA (FR).

ACKNOWLEDGMENT
The AGLAEA project is supported by the European Commission
through the Sixth Research Framework Programme for Research
and Technological Development (FP6, 2002-2006). The AGLAEA
project addresses the thematic area “Life sciences, genomics and
biotechnology for health”. The project started on 1st January 2007
and will last 36 months.

CONTACT
Project Coordinator With the support of ALMA Consulting Group
ADDEX Pharmaceuticals - Dr. Emmanuel Le Poul: Dr. Raffaella Catena:
Emmanuel.LePoul@addexpharma.com rcatena@almacg.com

Dissemination Manager
UNOTT University of Nottingham - Prof. Charles Marsden:
Charles.Marsden@nottingham.ac.uk

www.aglaea.eu