Acetylcholine controls GABA-, glutamate-, and glycine-dependent giant depolarizing potentials that govern spontaneous motoneuron activity at the onset of synaptogenesis in the mouse embryonic spinal cord

J Neurosci. 2014 Apr 30;34(18):6389-404. doi: 10.1523/JNEUROSCI.2664-13.2014.


A remarkable feature of early neuronal networks is their endogenous ability to generate spontaneous rhythmic electrical activity independently of any external stimuli. In the mouse embryonic SC, this activity starts at an embryonic age of ∼ 12 d and is characterized by bursts of action potentials recurring every 2-3 min. Although these bursts have been extensively studied using extracellular recordings and are known to play an important role in motoneuron (MN) maturation, the mechanisms driving MN activity at the onset of synaptogenesis are still poorly understood. Because only cholinergic antagonists are known to abolish early spontaneous activity, it has long been assumed that spinal cord (SC) activity relies on a core network of MNs synchronized via direct cholinergic collaterals. Using a combination of whole-cell patch-clamp recordings and extracellular recordings in E12.5 isolated mouse SC preparations, we found that spontaneous MN activity is driven by recurrent giant depolarizing potentials. Our analysis reveals that these giant depolarizing potentials are mediated by the activation of GABA, glutamate, and glycine receptors. We did not detect direct nAChR activation evoked by ACh application on MNs, indicating that cholinergic inputs between MNs are not functional at this age. However, we obtained evidence that the cholinergic dependency of early SC activity reflects a presynaptic facilitation of GABA and glutamate synaptic release via nicotinic AChRs. Our study demonstrates that, even in its earliest form, the activity of spinal MNs relies on a refined poly-synaptic network and involves a tight presynaptic cholinergic regulation of both GABAergic and glutamatergic inputs.

Keywords: embryo; giant depolarizing potential; motoneuron; mouse; spinal cord; synaptogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholine / metabolism*
  • Acetylcholine / pharmacology
  • Action Potentials / drug effects
  • Action Potentials / physiology*
  • Animals
  • Cholinergic Agents / pharmacology
  • Embryo, Mammalian
  • Excitatory Amino Acid Agents / pharmacology
  • Female
  • Gap Junctions / drug effects
  • Gap Junctions / metabolism
  • Gap Junctions / physiology*
  • Glutamic Acid / metabolism*
  • Glutamic Acid / pharmacology
  • Glycine / metabolism*
  • Glycine / pharmacology
  • Homeodomain Proteins / genetics
  • In Vitro Techniques
  • Mice
  • Mice, Transgenic
  • Motor Neurons / drug effects
  • Motor Neurons / physiology*
  • Nerve Net / drug effects
  • Nerve Net / physiology*
  • Pregnancy
  • Spinal Cord / cytology*
  • Tetrodotoxin / pharmacology
  • Transcription Factors / genetics
  • gamma-Aminobutyric Acid / metabolism*
  • gamma-Aminobutyric Acid / pharmacology


  • Cholinergic Agents
  • Excitatory Amino Acid Agents
  • Homeodomain Proteins
  • Transcription Factors
  • Hb9 protein, mouse
  • Glutamic Acid
  • Tetrodotoxin
  • gamma-Aminobutyric Acid
  • Acetylcholine
  • Glycine