Research objectives and content Pax-6 gene is responsible for the small eye (Sey) mouse mutation. Previous studies suggest cortex malformations in the developing brain of Sey/Sey mice. The aim of the project is to investigate the mechanism which is disrupted during cortical neurogenesis in the Sey mouse. I will try to study in vitro the migration of neural cells in cortical development by developing an organ culture system using brain slices from mouse embryos. With this approach, the analysis of neurogenesis may be extended to stages several days after birth and we will be able to follow the fate of neurons in Sey mice. The process of cell migration can be followed directly over time by labelling cells using a marker gene which when expressed is non toxic and detectable in living cells. I will use two reporter genes, the gene encoding the jellyfish green fluorescent protein (GFP) and the bacterial LacZ gene. I will use lipophilic dyes, viral and retroviral vectors to label cells. The premigratory cells of the ventricular zone are a population susceptible to retroviral infection and I will use retroviral vectors to infect these proliferating cells. In addition adenovirus vectors will be use to infect not only mitotic but also postmitotic cells, labelling cells in any part of the brain. Training content (objective, benefit and expected impact)
This methodology will permit examination of the migration pattern of neuronal cells of Sey/Sey as compared to wildtype cells. The proposal try to answer some fundamental questions about the migratory route of affected neurons and the role that Pax-6 has in this deffects. In addition the proposed proyect will give me the opportunity to adquired experience in many embryological techniques required in the analysis of development and background in this area.
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