Muscarinic-2 and orexin-2 receptors on GABAergic and other neurons in the rat mesopontine tegmentum and their potential role in sleep-wake state control

J Comp Neurol. 2008 Oct 20;510(6):607-30. doi: 10.1002/cne.21803.


Acetylcholine (ACh) plays an important role in the promotion of paradoxical sleep (PS) with muscle atonia through the muscarinic-2 receptor (M2R) in the mesopontine tegmentum. Conversely, orexin (Orx or hypocretin) appears to be critical for the maintenance of waking with muscle tone through the orexin-2 (or hypocretin-B) receptor (Orx2R), which is lacking in dogs having narcolepsy with cataplexy. In dual-immunostained material viewed under fluorescence microscopy, we examined the presence and distribution of M2R or Orx2R labeling on all neuronal nuclei (NeuN)-stained neurons or on glutamic acid decarboxylase (GAD)-stained neurons through the mesopontine tegmentum. Applying stereological analysis, we determined that many neurons bear M2Rs on their membrane ( approximately 6,300), including relatively large, non-GABAergic cells, which predominate (>75%) in the oral and caudal pontine (PnO and PnC) reticular fields, and small, GABAergic cells ( approximately 2,800), which predominate (>80%) in the mesencephalic (Mes) reticular formation. Many neurons bear Orx2Rs on their membrane ( approximately 6,800), including relatively large, non-GABAergic cells, which predominate (>70%) through all reticular fields, and comparatively few GABAergic cells ( approximately 700). In triple-immunostained material viewed by confocal microscopy, many large neurons in PnO and PnC appear to bear both M2Rs and Orx2Rs on their membrane, indicating that ACh and Orx could exert opposing influences of inhibition vs. excitation on putative reticulo-spinal neurons and thus attenuate vs. facilitate activity and muscle tone. A few GABAergic cells bear both receptors and could as PS inhibitor neurons serve under these different influences to control PS effector neurons and accordingly gate PS and muscle atonia appropriately across sleep-wake states.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholine / metabolism
  • Animals
  • Biomarkers / metabolism
  • Cats
  • Dogs
  • Glutamate Decarboxylase / metabolism
  • Humans
  • Male
  • Narcolepsy / metabolism
  • Narcolepsy / physiopathology
  • Neurons / cytology
  • Neurons / metabolism*
  • Orexin Receptors
  • Pons / cytology
  • Pons / metabolism
  • Rats
  • Rats, Wistar
  • Receptor, Muscarinic M2 / metabolism*
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Neuropeptide / metabolism*
  • Sleep / physiology*
  • Tegmentum Mesencephali / cytology*
  • Tegmentum Mesencephali / metabolism
  • Vesicular Acetylcholine Transport Proteins / metabolism
  • Wakefulness / physiology*
  • gamma-Aminobutyric Acid / metabolism*


  • Biomarkers
  • Orexin Receptors
  • Receptor, Muscarinic M2
  • Receptors, G-Protein-Coupled
  • Receptors, Neuropeptide
  • Slc18a3 protein, rat
  • Vesicular Acetylcholine Transport Proteins
  • gamma-Aminobutyric Acid
  • Glutamate Decarboxylase
  • Acetylcholine