Visualizing and Electrophysiologically Characterizing Pyramidal Neurons in vmPFC

mCherry⁺ (Supplementary Fig. 11) or eYFP⁻ (Fig. 7) pyramidal neurons in layer 2/3 of vmPFC were visualized via infrared oblique-illumination (Olympus X51W; Hamamatsu CCD camera C11440) and randomly selected for voltage-clamp recordings. Patch pipettes (4–6 MΩ) were made from borosciliate glass using a micropipette puller (Model P-1000; Sutter Instruments). These pipettes were filled with pipette solution contained (in mM): 132.5 Cs-gluconate, 17.5 CsCl, 2 MgCl₂, 0.5 EGTA, 10 HEPES, 4 ATP, 5 QX-314 (pH 7.3). Evoked inhibitory post-synaptic currents (eIPSCs) were recorded by voltage clamping at +10 mV^{68 (link)} using an Axonpatch-700B amplifier (Axon Instruments). For optogenetic stimulation of ChR2-eYFP⁺ nNOS-expressing neurons in vmPFC, Single light pulse (5 ms duration) was delivered to the slice through an optical fiber from a 465 nm wavelength blue laser. All recordings were performed in the presence of TTX (0.5 μM) and 4-AP (100 μM) to obtain monosynaptic transmission.