0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

Sezione III: Attività per progetti
with a component that modulates it. One such protein is hexokinase, which
associates with the PTP, maintaining it closed. We will investigate whether
mutant Htt displaces hexokinase from the PTP, leading to its unwanted opening.
A relatively simple experiment will be to use specific antibodies to test the
amount of hexokinase bound to mitochondria in cells transfected with the 82-
Q Htt construct compared to control cells. The work will be then extended to
other approaches, such as the direct demonstration of the huntingtinhexokinase
interaction. Although hexokinase will be the preferred target in the first
phase of the study, we also plan to explore other known modulators of the
PTP, e.g., the adenine nucleotide translocator.
Furthermore, we will use the 82-Q construct to transfect the Stdh-Q7 control
cells. These cells, even if not yet mature neurons, are immortalized, and
will thus allow to study all developmental phases of the disease, from the presymptomatic
stage to the stage of apoptotic cell demise. Finally, the last part
of the HD project – which will be developed in parallel with the work
described above in the 2 nd and 3 rd year – will be carried out using the mouse
line YAC 128 that has now become commercially available. This line expresses
a full-length mutated Htt (all other HD mouse models express only the first
exon) and develops disease in a period of months, up to the death of the mice.
The experimental work described above will be replicated on primary cultures
of striatal neurons obtained from the YAC 128 line at all stages of disease progression.
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618 2009