Ph.D. Projects
Analysis of the Role of Ras-GRF1 and Ras-GRF2 in regulating Ras-ERK-dependent signalling in striatum-dependent behavioural plasticity by lentiviral vector assisted trangenesis and RNA interference
The striatum is a brain region in which a convergent activity of both glutamate- and dopamine- dependent cell signalling is essential for a variety of behavioural responses and neurological disorders, including procedural and motor learning as well as drug addiction, Parkinson’s and Huntington’s diseases. Others and we have shown that the ERK signalling pathway is implicated in the control of gene expression in the striatum, both during procedural learning and in response to drugs of abuse. Particularly important in the striatum appears to be the activity of a specific GDP-GTP exchange factor for Ras, Ras-GRF1, which acts as coincidence detector for both glutamate and dopamine dependent signals, the two main striatal inputs.
In order to investigate the role of Ras-GRF1 and its close homologue, Ras-GRF2, in the striatum, the PhD student will use viral constructs expressing dominant negative signalling mutants for these molecules. Thus, the achieved expression with a tight spatio-temporal control in intact mice, upon stereotactic injection in a striatum and under the control of the fully reversible TET system, will affect the ERK pathway in individual cells on demand, leaving nearby cells unaffected. Moreover, the PhD student will develop new lentiviral vectors to specifically “knock down” by RNA interference the two Ras-GRF forms, either individually or in combination in the striatum, to perform a complementary analysis. Mice will then be subjected to a battery of behavioural tests to evaluate the physiological effect of ERK blockade by Ras-GRF1/2 manipulation in the striatum. Treatments with drugs of abuse will also be included in the analysis.
