Analysis of NMDAR subunits expression after GluN2A knockdown in mature primary neuronal cultures

ACUTAIN, M. FLORENCIA; BAEZ, MARÍA VERÓNICA; JERUSALINSKY, DIANA ALICIA; VAZQUEZ, CECILIA

Mar del Plata

Congreso; XXXII Congress of the Argentine Society for Research in Neuroscience; 2017

Sociedad Argentina de Neurociencia

For several years, NMDA receptors (NMDAR) havebeen investigated through different approaches because of their role insynaptic plasticity, learning processes and memory. NMDAR are composed by twoGluN1 obligatory subunits and two regulatory subunits: GluN2 (A-D) or GluN3(A-B). In hippocampus and other memory related brain structures GluN2A andGluN2B are the most expressed regulatory subunits, with different expressionpatterns. While GluN2B is expressed in immature synapses, GluN2A is characteristicof mature and stable synapses. In order to understand the role of GluN2A duringmemory acquisition and plasticity induction we built two AAV-eGFP vectors: oneof them codifying a shRNA anti GluN2A (AAV-sh2A), and the other carrying ashRNA scramble as control (AAV-shSc). In this work we analyzed the specifity ofGluN2A knockdown in primary neuronal cultures infected with AAV-sh2A orAAV-shSc. We observed a decrease in GluN2A mRNA by qPCR only in primarycultures infected with AAVsh2A, without changes in GluN1 or GluN2B expression.Interestingly in those cultures, GluN2A decreased expression was accompanied bya significant diminution on GluN1 protein level. On the other hand, GluN2B levelswere similar to control cultures infected with the AAV-shSc. These resultssuggest that GluN2A decrease does not change NMDAR subunit expression at transcriptionlevel. However GluN2A decayed levels could activate some postranscriptionalregulatory mechanisms that change GluN1 protein levels.