Characterization of ena/VASP domains with dominant negative function

HERNÁNDEZ, MV; FRANCO, DL; CERIANI, MF

Huerta Grande

Congreso; XXVII Congreso de la Sociedad Argentina de Neurociencias; 2011

Sociedad Argentina de Neurociencias

Neurodegeneration is a process known to occur in metazoans. In previous work we found that the gene enabled (ena) is implicated in progressive neurodegeneration in Drosophila. Silencing ena ortholog genes (the ena/VASP family) in mouse hippocampal neurons triggered neurite retraction and concominant neuronal cell death through an apoptotic pathway. Since our ultimate goal is to confirm these results in a mouse model of late onset progressive neurodegeneration through deregulation of ena/VASP family members, we characterized potential dominant negative (DN) versions to identify the most effective one. Ena/VASP proteins share three well-defined domains, the EVH1 domain, a central proline-rich region, and EVH2. Overexpression of the DN (EVH1-GFP or EVH2-GFP) in mouse hipocampal neurons led to neurite retraction reminiscent of what had been observed with the RNAi pools. Thus, to begin to dissect potential signaling pathways we seek to define if a specific domain of ENA is responsible for the degenerative phenotype in Drosophila; transgenic fly lines expressing the EVH1 and EVH2 domains were generated. Preliminary results suggest that the functional inhibition of the EVH2 domain could be responsible for the behavioral phenotype observed in the original mutant.