Molecular Changes in GluN2A Knockdown of Mature Primary Neuronal Cultures

CECILIA A VAZQUEZ; MA. VERONICA BAEZ; MARIA FLORENCIA ACUTAIN; DIANA A. JERUSALINSKY; ANNA SALVETTI

Cordoba

Congreso; XXXIII Reunión Anual de la Sociedad Argentina de Investigacion en Neurociencias; 2018

Sociedad Argentina de Investigacion en Neurociencias

NMDA receptors (NMDAR) are glutamatergic receptors involved in synaptic plasticity, learning, and memory processes as well as in several neuropathologies. NMDAR are composed by two GluN1 obligatory subunits and two regulatory subunits: GluN2 (A-D) or GluN3 (A-B). In cognitive-related brain structures, GluN2A and GluN2B are the most expressed regulatory subunits, that undergoes a tightly regulation at transcriptional and translational level. Whereas GluN2B expression is characteristic of immature synapses, GluN2A is present in mature and stable synapses. In order to better understand the role of GluN2A in synapsis, we transduced mature neuronal cultures with AAV-eGFP vectors: one codifying a specific shRNA anti GluN2A, AAVsh2A, and the other carrying a shRNA scramble as control, AAV-shSc. As we verified that AAVsh2A knockdown GluN2A mRNA and protein levels (GluN2A KD), we analyzed the other NMDAR subunit expression in this cultures, as well as REST, a transcription factor that regulates GluN2A/GluN2B relationship. The GluN2A KD induced a decrease in REST levels without significant changes in GluN2B expression. On the other hand, GluN1 protein levels were significantly low in GluN2A KD cultures in spite of control mRNA levels. Furthermore, GluN1 splicing variants proportion was altered. These results suggest that GluN2A KD induce a rearrangement of NMDAR and REST expression similar to those observed in more immature states at neuronal differentiation.