0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

HSR.1 – Effects of laquinimod on synaptic neurodegenerative damage in experimental MS
a) By studying if Laquinimod treatment is able to decrease the loss of
myelin and viable axons in the forebrain and in the spinal cord of EAE mice.
b) By studying if Laquinimod treatment is able to restore disrupted SVZ
and change the fate of endogenous NSC interfering with their differentiation
program during ongoing EAE.
c) By studying if Laquinimod treatment is able to prevent early synaptopathy
in EAE mice.
d) By investigating whether Laquinimod is able to affect excitatory and
inhibitory synaptic transmission independently of its action on microglia or T
cells.
The final goal of the study is to provide a reliable mechanism at the basis
of the putative neuroprotective action of Laquinimod in experimental MS. We
will also be in the position to understand whether the neuroprotective effects
of Laquinimod are immuno-mediated or are secondary to a direct effect on
neuronal elements of the CNS.
METHODS
We will induce EAE in C57BL/6 mice by immunization with MOG35-55
in complete Freund’s adjuvant [Pluchino et al. 2003]. Laquinimod will be
administered to EAE mice according to published protocols [Brunmark et al.
2002; Aharoni et al. 2005]. Details of Laquinimod administration are specified
in Table 1.
Table 1
N. Treatement Schedule Route Dose
10 + 10 Laquinimod every other day from day +1 oral 1mg/kg
10 + 10 Laquinimod every other day from day +1 oral 5mg/kg
15 + 15 Laquinimod every other day from day +1 oral 10mg/kg
15 + 15 PBS every other day from day +1 oral 200microl/mice
EAE score and body weight will be recorded daily.
Analyses will include:
Aim a – Transcardially perfused mice will be processed for neuropathological
studies including semi-quantitative evaluation of inflammatory infiltrates,
demyelination, and axonal loss on both spinal cord and fore-brain,
namely the striatum.
Aim b – The dating of neural stem/precursor cells will be studied using
labeling-retaining techniques able to detect cells undergoing DNA synthesis
(e.g. BrdU, IddU) [Takahashi et al. 1992] combined with techniques able to
detect cell markers (e.g. immunofluorescence). If positive results will be
obtained, we will further dissect the fate of NSCs in a second set of experi-
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