Neurons generated by direct conversion of fibroblasts reproduce synaptic phenotype caused by autism-associated neuroligin-3 mutation

Proc Natl Acad Sci U S A. 2013 Oct 8;110(41):16622-7. doi: 10.1073/pnas.1316240110. Epub 2013 Sep 17.

Abstract

Recent studies suggest that induced neuronal (iN) cells that are directly transdifferentiated from nonneuronal cells provide a powerful opportunity to examine neuropsychiatric diseases. However, the validity of using this approach to examine disease-specific changes has not been demonstrated. Here, we analyze the phenotypes of iN cells that were derived from murine embryonic fibroblasts cultured from littermate wild-type and mutant mice carrying the autism-associated R704C substitution in neuroligin-3. We show that neuroligin-3 R704C-mutant iN cells exhibit a large and selective decrease in AMPA-type glutamate receptor-mediated synaptic transmission without changes in NMDA-type glutamate receptor- or in GABAA receptor-mediated synaptic transmission. Thus, the synaptic phenotype observed in R704C-mutant iN cells replicates the previously observed phenotype of R704C-mutant neurons. Our data show that the effect of the R704C mutation is applicable even to neurons transdifferentiated from fibroblasts and constitute a proof-of-concept demonstration that iN cells can be used for cellular disease modeling.

Keywords: cellular reprogramming; neurexin; postsynaptic density; stem cells; synapse.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution / genetics
  • Animals
  • Autistic Disorder / genetics
  • Autistic Disorder / physiopathology*
  • Cell Adhesion Molecules, Neuronal / genetics*
  • Cell Transdifferentiation / physiology
  • Cells, Cultured
  • Disease Models, Animal*
  • Fibroblasts / cytology*
  • Fibroblasts / physiology
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Membrane Proteins / genetics*
  • Mice
  • Nerve Tissue Proteins / genetics*
  • Neurons / cytology*
  • Patch-Clamp Techniques
  • Phenotype*
  • Real-Time Polymerase Chain Reaction
  • Receptors, AMPA / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Synaptic Transmission / genetics
  • Synaptic Transmission / physiology

Substances

  • Cell Adhesion Molecules, Neuronal
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Receptors, AMPA
  • neuroligin 3