The siRNA-mediated knockdown of GluN3A in 46C-derived neural stem cells affects mRNA expression levels of neural genes, including known iGluR interactors

PLoS One. 2018 Feb 13;13(2):e0192242. doi: 10.1371/journal.pone.0192242. eCollection 2018.


For years, GluN3A was solely considered to be a dominant-negative modulator of NMDARs, since its incorporation into receptors alters hallmark features of conventional NMDARs composed of GluN1/GluN2 subunits. Only recently, increasing evidence has accumulated that GluN3A plays a more diversified role. It is considered to be critically involved in the maturation of glutamatergic synapses, and it might act as a molecular brake to prevent premature synaptic strengthening. Its expression pattern supports a putative role during neural development, since GluN3A is predominantly expressed in early pre- and postnatal stages. In this study, we used RNA interference to efficiently knock down GluN3A in 46C-derived neural stem cells (NSCs) both at the mRNA and at the protein level. Global gene expression profiling upon GluN3A knockdown revealed significantly altered expression of a multitude of neural genes, including genes encoding small GTPases, retinal proteins, and cytoskeletal proteins, some of which have been previously shown to interact with GluN3A or other iGluR subunits. Canonical pathway enrichment studies point at important roles of GluN3A affecting key cellular pathways involved in cell growth, proliferation, motility, and survival, such as the mTOR pathway. This study for the first time provides insights into transcriptome changes upon the specific knockdown of an NMDAR subunit in NSCs, which may help to identify additional functions and downstream pathways of GluN3A and GluN3A-containing NMDARs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Gene Expression Profiling
  • Gene Knockdown Techniques*
  • Mice
  • Neural Stem Cells / metabolism*
  • Protein Binding
  • RNA, Messenger / genetics*
  • RNA, Small Interfering / genetics*
  • Receptors, N-Methyl-D-Aspartate / genetics*
  • Receptors, N-Methyl-D-Aspartate / metabolism


  • GluN3A protein, mouse
  • RNA, Messenger
  • RNA, Small Interfering
  • Receptors, N-Methyl-D-Aspartate

Grant support

SP and EE received PhD scholarships from the International Graduate School of Neuroscience, Ruhr University Bochum, Germany ( and the Ruhr University Research School, Ruhr University Bochum, Germany ( The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.