Ph.D. Projects
Project (P.I. Gianvito Martino). Neural stem/progenitor cells (NPCs) are central in the regenerating processes occurring within the central nervous system (CNS) as a consequence of inflammatory and degenerative insults. In this project we aim at investigating the putative reparative role of NPCs in inflammatory and neurodegenerative disorders of the CNS by using a recently developed transgenic tool in which the selective killing of NPCs is possible. We generated a NestfloxGFPfloxTK-IRES-LacZ transgenic mouse line by using the lentiviral technology; the construct consists in a loxP flanked GFP gene placed under the control of Nestin regulatory regions. In these mice three stop codons were placed downstream the GFP gene, so that the TK gene were translated only after a removal of the GFP segment. In order to perform the selective killing of NPCs NestfloxGFPfloxTK-IRES-LacZ transgenic mouse line were crossed to CMVCre transgenic mice. Using this mouse line, we aim at: (i) understanding the physiological role of NPCs; (ii) understanding the functional role of NPCs in inflammatory and neurodegenerative disorders affecting the CNS, such as experimental multiple sclerosis and stroke.
Project (P.I. Luca Muzio). Microglia play a key role in both detrimental and reparative processes occurring within the central nervous system (CNS) as a consequence of inflammatory and degenerative insults. In this project we aim at investigating the molecular mechanisms underlying the detrimental role exerted by microglia on synaptic activity within the CNS which has been recently described in mice affected by experimental autoimmune encephalomyelitis (EAE), a mouse model of a CNS-confined disease encompassing inflammatory as well as degenerative pathological features. A genetic screening will be first performed in EAE mice by comparing CNS affected vs. non affected CNS areas so to assess the precise composition of the putative detrimental pro-inflammatory milieu. Genes coding for soluble pro-inflammatory molecules released by microglia and their relationship with genes coding for molecules regulating the synaptic strength will be then identified. Once candidate molecules will be identified, their effects on neuronal function will be assayed using Micro Electrode Array (MEA) devices in order to identify molecular patterns concurring to neuronal degeneration. Finally, the relevance of such molecules in controlling the neuronal derangement will be functionally proved in vivo using lentiviral mediated transgenesis.
