Evidence for M muscarinic receptor modulation of axon terminals and dendrites in the rodent basolateral amygdala: An ultrastructural and electrophysiological analysis.

The basolateral amygdala receives a very dense cholinergic innervation from the basal forebrain that is important for memory consolidation. Although behavioral studies have shown that both M and M muscarinic receptors are critical for these mnemonic functions, there have been very few neuroanatomical and electrophysiological investigations of the localization and function of different types of muscarinic receptors in the amygdala. In the present study we investigated the subcellular localization of M muscarinic receptors (MRs) in the anterior basolateral nucleus (BLa) of the mouse, including the localization of MRs in parvalbumin (PV) immunoreactive interneurons, using double-labeling immunoelectron microscopy. Little if any MR-immunoreactivity (MR-ir) was observed in neuronal somata, but the neuropil was densely labeled. Ultrastructural analysis using a pre-embedding immunogold-silver technique (IGS) demonstrated MR-ir in dendritic shafts, spines, and axon terminals forming asymmetrical (excitatory) or symmetrical (mostly inhibitory) synapses. In addition, about one-quarter of PV+ axon terminals and half of PV+ dendrites, localized using immunoperoxidase, were MR+ when observed in single thin sections. In all MR+ neuropilar structures, including those that were PV+, about one-quarter to two-thirds of MR+ immunoparticles were plasma-membrane-associated, depending on the structure. The expression of MRs in PV+ and PV-negative terminals forming symmetrical synapses indicates MR modulation of inhibitory transmission. Electrophysiological studies in mouse and rat brain slices, including paired recordings from interneurons and pyramidal projection neurons, demonstrated MR-mediated suppression of GABA release. These findings suggest cell-type-specific functions of MRs and shed light on organizing principles of cholinergic modulation in the BLa.