Neuroligin-4 Regulates Excitatory Synaptic Transmission in Human Neurons

Neuron. 2019 Aug 21;103(4):617-626.e6. doi: 10.1016/j.neuron.2019.05.043. Epub 2019 Jun 27.


The autism-associated synaptic-adhesion gene Neuroligin-4 (NLGN4) is poorly conserved evolutionarily, limiting conclusions from Nlgn4 mouse models for human cells. Here, we show that the cellular and subcellular expression of human and murine Neuroligin-4 differ, with human Neuroligin-4 primarily expressed in cerebral cortex and localized to excitatory synapses. Overexpression of NLGN4 in human embryonic stem cell-derived neurons resulted in an increase in excitatory synapse numbers but a remarkable decrease in synaptic strength. Human neurons carrying the syndromic autism mutation NLGN4-R704C also formed more excitatory synapses but with increased functional synaptic transmission due to a postsynaptic mechanism, while genetic loss of NLGN4 did not significantly affect synapses in the human neurons analyzed. Thus, the NLGN4-R704C mutation represents a change-of-function mutation. Our work reveals contrasting roles of NLGN4 in human and mouse neurons, suggesting that human evolution has impacted even fundamental cell biological processes generally assumed to be highly conserved.

Keywords: ASD; Neuroligin-4; autism; embryonic stem cells; induced neuronal (iN) cells; isogenic cell lines; synaptic transmission.

Publication types

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

MeSH terms

  • Animals
  • Autistic Disorder / genetics
  • Cell Adhesion Molecules, Neuronal / genetics
  • Cell Adhesion Molecules, Neuronal / physiology*
  • Cells, Cultured
  • Cerebral Cortex / physiology
  • Embryonic Stem Cells / cytology
  • Excitatory Postsynaptic Potentials / physiology
  • Genes, Reporter
  • Glutamic Acid / physiology
  • Humans
  • Mice
  • Miniature Postsynaptic Potentials / physiology
  • Mutation, Missense
  • Neurogenesis
  • Neurons / physiology
  • Phenotype
  • Receptors, Glutamate / physiology
  • Species Specificity
  • Synapses / chemistry
  • Synaptic Transmission / physiology*


  • Cell Adhesion Molecules, Neuronal
  • NLGN4X protein, human
  • Nlgn4 protein, mouse
  • Receptors, Glutamate
  • alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid subtype glutamate receptor, human
  • Glutamic Acid