Nitric Oxide Signaling Strengthens Inhibitory Synapses of Cerebellar Molecular Layer Interneurons through a GABARAP-Dependent Mechanism

J Neurosci. 2020 Apr 22;40(17):3348-3359. doi: 10.1523/JNEUROSCI.2211-19.2020. Epub 2020 Mar 13.


Nitric oxide (NO) is an important signaling molecule that fulfills diverse functional roles as a neurotransmitter or diffusible second messenger in the developing and adult CNS. Although the impact of NO on different behaviors such as movement, sleep, learning, and memory has been well documented, the identity of its molecular and cellular targets is still an area of ongoing investigation. Here, we identify a novel role for NO in strengthening inhibitory GABAA receptor-mediated transmission in molecular layer interneurons of the mouse cerebellum. NO levels are elevated by the activity of neuronal NO synthase (nNOS) following Ca2+ entry through extrasynaptic NMDA-type ionotropic glutamate receptors (NMDARs). NO activates protein kinase G with the subsequent production of cGMP, which prompts the stimulation of NADPH oxidase and protein kinase C (PKC). The activation of PKC promotes the selective strengthening of α3-containing GABAARs synapses through a GΑΒΑ receptor-associated protein-dependent mechanism. Given the widespread but cell type-specific expression of the NMDAR/nNOS complex in the mammalian brain, our data suggest that NMDARs may uniquely strengthen inhibitory GABAergic transmission in these cells through a novel NO-mediated pathway.SIGNIFICANCE STATEMENT Long-term changes in the efficacy of GABAergic transmission is mediated by multiple presynaptic and postsynaptic mechanisms. A prominent pathway involves crosstalk between excitatory and inhibitory synapses whereby Ca2+-entering through postsynaptic NMDARs promotes the recruitment and strengthening of GABAA receptor synapses via Ca2+/calmodulin-dependent protein kinase II. Although Ca2+ transport by NMDARs is also tightly coupled to nNOS activity and NO production, it has yet to be determined whether this pathway affects inhibitory synapses. Here, we show that activation of NMDARs trigger a NO-dependent pathway that strengthens inhibitory GABAergic synapses of cerebellar molecular layer interneurons. Given the widespread expression of NMDARs and nNOS in the mammalian brain, we speculate that NO control of GABAergic synapse efficacy may be more widespread than has been appreciated.

Keywords: GABA receptor; GABARAP; cerebellum; electrophysiology; inhibitory synapse; plasticity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins / metabolism*
  • Cerebellum / drug effects
  • Cerebellum / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Excitatory Amino Acid Antagonists / pharmacology
  • Female
  • Interneurons / drug effects
  • Interneurons / metabolism*
  • Male
  • Mice
  • Microtubule-Associated Proteins / metabolism*
  • Neural Inhibition / drug effects
  • Neural Inhibition / physiology
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase Type I / antagonists & inhibitors
  • Patch-Clamp Techniques
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Synaptic Transmission / drug effects
  • Synaptic Transmission / physiology


  • Apoptosis Regulatory Proteins
  • Enzyme Inhibitors
  • Excitatory Amino Acid Antagonists
  • GABARAP protein, mouse
  • Microtubule-Associated Proteins
  • Nitric Oxide
  • Nitric Oxide Synthase Type I