Titolo della tesi: Investigating the extrinsic niche molecular pathways regulating mouse neural/stem progenitor cell proliferation and and survival
In my PhD course I was focused basically on the study of neural stem/progenitor cells (NSPCs) derived from the mouse cerebral cortex at the embryonic 13.5 day of development (Ctx NSPCs), when a peak in neurogenesis occurs 1. In particular, I was interested in the mechanisms that regulate NSPC cell cycle. The main theme of my research was the characterization of the effects of PF3845, an irreversible inhibitor of fatty acid amide hydrolase (FAAH) on NSPCs. FAAH is the main degrading enzyme of Anandamide (AEA), one of the most characterised endocannabinoids produced in the brain, and of its analogues, Palmithoylethanolamide (PEA) and Oleoylethanolamide (OEA), two endocannabinoid-like compounds. These three compounds belong to the N-acylethanolamine family, a class of bioactive lipids well-known for acting on inflammation, acute and chronic pain, obesity and central nervous system diseases but their role in neurogenesis needs to be deeply investigated. I studied the effects of the inhibition of FAAH in Ctx NSPCs grown in proliferating culture conditions. During my PhD course, I also participated in a project of my laboratory team aimed at investigating the dose-dependent and time-dependent response of Ctx NSPCs to X-ray irradiation. For both projects, we took advantage of a well-established in vitro NSPC culture system. Morphological, immunofluorescent and molecular analyses and flow cytometry assays were utilised to evaluate cell viability and proliferation capacity. As part of my PhD, I spent four months in a foreign laboratory at the Biomedical Research Institute of Málaga (IBMA), where I improved my technical skills in protein analysis. In particular, I studied the endocannabinoid system in an amyloid mouse model of AD (5xFAD) submitted to a western diet, enriched in fat content.