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Fig. 2 ACh depletion from the vMHb impairs neural transmission to the IPN (A) Schematic illustration of preparation for slice containing the entire vMHb-fr-IPN pathway. (B) Distribution of the Hb, the fr, and the IPN in the paracoronal brain slice (red box in A) used for electrophysiological recording and calcium imaging. Scale bar, 200 μm. (C) EPSCs evoked by 20-s 20-Hz tetanic stimulation to the left fr. Six traces are averaged (Cont, black; cCKD, red). Gray traces indicate the superimposition of all six traces. (D) Slow components of EPSCs of Cont (black, n = 6) and cCKD (red, n = 6) slices. Gray (Cont) and pale red (cCKD) traces indicate SEM. Bottom panel indicates the postsynaptic charge transfer (integration of slow EPSCs during 20-s stimulation) of Cont and cCKD slices (p = 0.0309). (E) Fast components of EPSCs of Cont (black) and cCKD (red) slices. Gray (Cont) and pale red (cCKD) traces indicate SEM. Bottom panel indicates the average of fast EPSC amplitude of Cont and cCKD slices (p > 0.05). Bars in (D) and (E), mean. Unpaired t test was used for statistical analyses. Statistical significance was defined as ∗p < 0.05.