Muscarinic signaling sustains delay-period selectivity in prefrontal cortex by preserving thalamic drive

Short-term memory (STM) requires the temporary maintenance of task-relevant information in prefrontal cortex (PFC). Reciprocal interactions between the mediodorsal thalamus (MD) and PFC have been implicated in sustaining this activity, but how thalamofrontal communication is maintained despite the biophysical constraints of repetitive synaptic transmission remains unclear. Here, we show that MD–PFC synapses exhibit profound frequency-dependent short-term depression, causing repetitive thalamic input to progressively lose its ability to drive PFC neurons. During STM performance, acetylcholine (ACh) levels in PFC rose rapidly following cue presentation and remained elevated throughout the delay period. Muscarinic ACh receptor (mAChR) activation did not alleviate synaptic depression, but instead increased the intrinsic excitability of PFC neurons, thereby preserving their responsiveness to weakened thalamic input. Conversely, mAChR blockade impaired STM performance and progressively disrupted delay-period directional representations at both the single-neuron and population levels, while largely sparing initial encoding. These findings identify muscarinic signaling as a postsynaptic gain mechanism that compensates for declining thalamofrontal efficacy and sustains task-relevant prefrontal representations during STM.