Analysis of the prion mechanism using immortalized prionless neuronal cell

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Prion is a cellular protein (PrPc) which has been implicated in transmissible neurodegenerative disease This protein is found on the outer surfaces of neurons of the brain. Although the PrPc function has still not been completely clarified, it seems to be physiologically important because its gene has been found in all mammal examined so far. The establishment of progenitor cell lines from Prn-p gene-target knock out mice embryos and from the wild type would be a valuable model system to clarify if PrPc is related with putative differences in the development of those two cell system and also to verify if there is presence of compensatory gene control mechanisms early in embryogenesis.Mice homozogous for disrupted Prn-p genes have been generated (Bueler et al. 1987).From this type of gene-targeted mice, hippocampal cells have been immortalized and clones were obtatined. The hippocampus has been dissected from brain of mouse embryos at embryonic day 14, as the PrPc is predominantly a neuronal protein, presented in a high proportion of hippocampus nerve cell, and that the neurogenesis in vivo are prefferentially occuring at early development.Twenty eight clones were obtained and has been characterized as prionless and neuronal origin.