IBCN   20355
INSTITUTO DE BIOLOGIA CELULAR Y NEUROCIENCIA "PROFESOR EDUARDO DE ROBERTIS"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Development of amplicon vectors as tools for silencing NMDA receptor subunits
Autor/es:
AGUIRRE AI; BAEZ MV; MELENDEZ M; ADROVER M; EPSTEIN AL; JERUSALINSKY DA
Lugar:
CABA
Reunión:
Simposio; 2do Simposio Franco Argentino de Neurociencias; 2012
Resumen:
N-Methyl-D-aspartate receptors (NMDAR) are involved in the physiology of synaptic plasticity, learning and memory and in the pathology of several CNS diseases. They are tetramers formed by two obligatory GluN1 subunits and different regulatory subunits. GluN2A and GluN2B are the main regulatory subunits in the forebrain. We have developed amplicon vectors derived from herpes simplex virus type-1 (HSV-1) to study the role of NMDAR, particularly those carrying GluN2A and GluN2B, in synaptic plasticity, learning and memory. Amplicon vectors were chosen as tools because of their tropism, large transgene capacity (up to 150 kb) and ability to maintain transgene expression. We have demonstrated that amplicon vectors bearing sequences for GluN1 subunit overexpressed it in vitro in primary cultures of cortical neurons. We have also shown that viral vectors carrying antisense sequences for GluN1 decreased its expression at the hippocampus in vivo and impaired performance of rats in hippocampal-depending behavioural tasks. Same vectors have been used to analize the binding of A-oligomers to neurons (Decker et al., 2010). We built up amplicon vectors carrying sequences encoding short-hairpin RNAs (shRNAs) against GluN1 or GluN2B, or antisense sequences for GluN2B, to produce a temporal and selective decrease in their expression. Their capacity to decrease the expression of the correspondent subunit was shown in primary cultures of hippocampal neurons. In the near future, we plan to assay these vectors in vivo, expecting to modify the expression of different GluN2 subunits in the rat hippocampus and to test, through behavioural tasks, the role of each receptor subtype in memory and in some early events of pathological processes.Supported by LIA DEVENIR (CONICET-UBA/CNRS-UCBL) and ECOS Sud-MinCyT A08S04.