Cognitive studies and direct cell reprogramming protocol for the aging rat.

LOPEZ-LEÓN M; MOREL GR; URIARTE DONATI MAIA; LEHMANN M; GOYA RG

Vancouver

Congreso; 9th Canadian Neuroscience Meeting 2015; 2015

International Brain Research Organization

We use the aging rat (AR) as a model of age-related cognitive decline and plan to use cell reprogramming as a restorative intervention in their hippocampus. We assessed the extent of spatial memory (SM) impairment in 26 (old), 31 (senile) and 5 (young) months female rats as well as the age-related changes in their dorsal hippocampus. Age changes in SM performance were assessed with the Barnes maze test. We employed two probe trials, 1 and 5 days after training, in order to evaluate "short-term" and "middle-term" SM. The results revealed that old rats perform better than senile rats in acquisition trials and young rats perform better than both aging groups. Morphological analysis of the dorsal hippocampus showed a marked decrease (94-97%) in doublecortin neuron number in the dentate gyrus in both aging groups. It has been recently demonstrated that transient induction of the four pluripotency genes oct4, sox2, klf4 and c-myc (the Yamanaka genes), followed by appropriate signalling inputs, can efficiently transdifferentiate fibroblasts into neural stem/progenitor cells (NPCs). We report here the ongoing construction of a helper-dependent recombinant adenovector for the simultaneous expression of hGFP and the Yamanaka genes all under the control of a bidirectional Tet-Off regulatable promoter. This vector will be used to implement direct cell reprogramming in a non-integrative fashion. We describe the direct reprogramming protocol that we plan to use to convert fibroblasts into NPCs, which will be subsequently used for experimental cell therapy in the hippocampus of AR.