Extended Data Fig. 1: Fasting-induced response of c-Fos expression between ArcAgRP and ArcGABA+ neurons and NaChBac expression increases neuron activity.
From: Profound and redundant functions of arcuate neurons in obesity development
a, Matched sections from Arc in serial rostral to caudal Bregma levels (mm) showing c-Fos (red) and GFP reporter for AgRP neurons (left panels) or ArcGABA+ neurons (right panels). The pictures shown in the middle are magnified pictures of the boxed area at each indicated Bregma level. 3 V: the third ventricle. This experiment was repeated in 4 animals/group as indicated in Fig. 1a. b–k, Brain slices from Vgat-Cre::Ai9 reporter mice with stereotaxic injections of Cre-dependent GFP or NachBac vectors to the Arc were used for recording. b, c, Representative traces showing action potential firing by current injections in a step wise increment of 5pA with voltage clamped at -70 mV in control (b) and NachBac expressing neurons (c). d-e, Comparison in Rheobase (minimum current size required to be injected to elicit AP firing) (d, t = 2.523, df=31, *p = 0.0170) and comparison in minimum membrane potential required to elicit AP firing (e, t = 2.436, df=25, *p = 0.0223). f, g, Representative traces showing Na+ current traces with step wise voltage depolarization at the +10 mV increment from -70mV in control (f) and NachBac expressing neurons (g). h–k, Comparison in Na+ currents at the indicated voltages (h, −60 mV: t = 3.507, df=30, **p = 0.0015; −50mV: t = 8.754, df=30, **p < 0.0001; −40 mV: t = 5.459, df=30, **p < 0.0001), in input resistance of the recorded neurons between groups (i, t = 5.200, df=32, **p < 0.0001), in number of recorded neurons showing spontaneous firing between groups (j) and in resting membrane potential (REM) between the two groups (k, t = 2.441, df=32, *p = 0.0204). For b–k, n = 15 (control) or n = 19 (NachBac), all with 2-sided paired Student’s t tests. 3 V: the third ventricle. All data presented as mean + /-sem.
