Trafficking of the muscarinic m2 autoreceptor in cholinergic basalocortical neurons in vivo: differential regulation of plasma membrane receptor availability and intraneuronal localization in acetylcholinesterase-deficient and -inhibited mice.

Abstract

In vivo, the abundance of receptors at the neuronal plasma membrane may be critical in the mediation of pre- and postsynaptic responses. Thus, we have studied the membrane availability and intraneuronal distribution of the m2 muscarinic autoreceptor (m2R) in cholinergic neurons of the nucleus basalis magnocellularis (NBM) projecting to the frontal cortex (FC). We have studied the subcellular compartmentalization of m2R at somatodendritic postsynaptic and axonal presynaptic sites in control animals (AChE +/+) and in two animal models: mice displaying acute acetylcholinesterase (AChE) inhibition by treatment with metrifonate, and AChE-deficient mice (AChE -/-). In control animals, m2R was mainly located at the plasma membrane in the somatodendritic field of NBM and in cortical varicosities. Acute AChE inhibition and chronic AChE deficiency induced a dramatic decrease of cell surface m2R in the somatodendritic compartment. This finding was associated with two different intracytoplasmic events: (1). internalization of m2R in endosomes after acute AChE inhibition, (2). exaggerated storage of m2R in the endoplasmic reticulum and Golgi complex in AChE -/- mice. In contrast, the m2R density was higher at the membrane of cortical varicosities in AChE -/- mice but unchanged in acutely AChE-inhibited mice. Our data demonstrate that acute and chronic stimulation provoke, in vivo, depletion of the membrane store of somatodendritic m2R through different intracellular mechanisms: endocytosis of receptors from the plasma membrane to the cytoplasm (acute) or regulation of their delivery from intracytoplasmic stores to the plasma membrane (chronic). The increase of m2R at the membrane of axonal varicosities after chronic stimulation suggest modulation of presynaptic cholinergic activity, including neurotransmitter release.