Extended Data Figure 6: Pomc expression is reduced in pair-fed DBB-ablated animals.

a, Relative Agrp and Pomc expression levels in arcuate nuclei between pair-fed non-ablated and DBB-ablated mice (n = 4 mice per group). Data are represented as mean ± s.e.m. *P < 0.05 by two-sided, unpaired Student’s t-test. b, Acetylcholine does not significantly alter arcuate NPY neuron firing. Data shown as mean ± s.e.m. of spike frequency in 8 acetylcholine-treated cells (1 recorded neuron per slice, 1–2 slices per mouse, n = 6 mice), comparisons made versus baseline values (1–4 min) by repeated measures ANOVA with Holm–Sidak multiple comparison. c, d, Acetylcholine significantly increases arcuate POMC neuron firing. Data shown as mean ± s.e.m. of spike frequency in 7 acetylcholine-treated cells (1 recorded neuron per slice, 1–2 slices per mouse, n = 4 mice), *P < 0.001 versus baseline values (1–4 min) by repeated measures ANOVA with Holm–Sidak multiple comparison. e, Representative voltage clamp recording from POMC-EGFP neuron after local administration of acetylcholine and after pharmacological manipulation of fast synaptic transmission, as well as nicotinic acetylcholine receptor blockade (20-s sweep with an inter-trial interval of 1 min and repeated for 5 sweeps each for baseline, synaptic blockers (CNQX, 2-amino-5-phosphonopentanoic acid (APV), GABAzine and tetrodotoxin (TTX)), and nicotinic blockers (mecamylamine (MEC), methyllycaconitine (MLA) and dihydro-β-erythroidine hydrobromide (DBHE)).