Synaptic Release of Acetylcholine Rapidly Suppresses Cortical Activity by Recruiting Muscarinic Receptors in Layer 4

Abstract

Cholinergic afferents from the basal forebrain (BF) can influence cortical activity on rapid time scales, enabling sensory information processing and exploratory behavior. However, our understanding of how synaptically released acetylcholine (ACh) influences cellular targets in distinct cortical layers remains incomplete. Previous studies have shown that rapid changes in cortical dynamics induced by phasic BF activity can be mediated by the activation of nicotinic ACh receptors (nAChRs) expressed in distinct types of GABAergic interneurons. In contrast, muscarinic ACh receptors (mAChRs) are assumed to be involved in slower and more diffuse ACh signaling following sustained increases in afferent activity. Here, we examined the mechanisms underlying fast cholinergic control of cortical circuit dynamics by pairing optical stimulation of cholinergic afferents with evoked activity in somatosensory cortical slices of mice of either sex. ACh release evoked by single stimuli led to a rapid and persistent suppression of cortical activity, mediated by mAChRs expressed in layer 4 and to a lesser extent, by nAChRs in layers 1-3. In agreement, we found that cholinergic inputs to layer 4 evoked short-latency and long-lasting mAChR-dependent inhibition of the large majority of excitatory neurons, whereas inputs to layers 1-3 primarily evoked nAChR-dependent excitation of different classes of interneurons. Our results indicate that the rapid cholinergic control of cortical network dynamics is mediated by both nAChRs and mAChRs-dependent mechanisms, which are expressed in distinct cortical layers and cell types. Acetylcholine (ACh) release from basal forebrain (BF) afferents to cortex influences a variety of cognitive functions including attention, sensory processing, and learning. Cholinergic control occurs on the time scale of seconds and is mediated by BF neurons that generate action potentials at low rates, indicating that ACh acts as a point-to-point neurotransmitter. Our findings highlight that even brief activation of cholinergic afferents can recruit both nicotinic and muscarinic ACh receptors expressed in several cell types, leading to modulation of cortical activity on distinct time scales. Furthermore, they indicate that the initial stages of cortical sensory processing are under direct cholinergic control.

Cortical ChAT+ neurons co-transmit acetylcholine and GABA in a target- and brain-region-specific manner

“…The existence of cortical ChAT + neurons requires a reevaluation of studies that globally manipulate cholinergic signaling in cortex. While many studies specifically targeted cortically-projecting basal forebrain neurons, several have used genetic crosses that affect all cholinergic neurons in the brain ( Chen et al, 2015 ; Sparks et al, 2017 ; Kuchibhotla et al, 2017 ; Dasgupta et al, 2018 ), and therefore include confounding effects from local VIP + /ChAT + interneurons. Studies that use ChAT-BAC-ChR2 mice to activate cholinergic neurons not only run the risk of confounding gain-of-function effects due to overexpressed VAChT ( Kolisnyk et al, 2013 ), but also from incidental manipulation of cortical VIP + interneurons, which are known to have profound effects on cortical function even purely through GABA release.…”

Cortical ChAT+neurons co-transmit acetylcholine and GABA in a target- and brain-region specific manner

“…The existence of cortical ChAT + neurons requires a reevaluation of studies that globally manipulate cholinergic signaling in cortex. While many studies specifically targeted cortically-projecting basal forebrain neurons, several have used genetic crosses that affect all cholinergic neurons in the brain 62–65 , and therefore include confounding effects from local VIP + /ChAT + interneurons. Studies that use ChAT-BAC-ChR2 mice to activate cholinergic neurons not only run the risk of confounding gain-of-function effects due to overexpressed VAChT 66 , but also from incidental manipulation of cortical VIP interneurons, which are known to have profound effects on cortical function even purely through GABA release.…”

Enhanced cortical responsiveness during natural sleep in freely behaving mice

“…Waking is characterized by high monoaminergic and cholinergic tone, SWS by low monoaminergic and cholinergic tone, and REM sleep by high cholinergic, but low monoaminergic tone 33 . Acetylcholine (ACh) inhibits principal neurons in layers II to V in the cortex 34 particularly in spiny stellate neurons and their recurrent network 35,36 . This might be a major contributor to the increased responses in SWS compared with waking.…”