Fig. 4: Systemic eNAMPT activity regulates hypothalamic POMC and NPY/AGRP neurons.

a Effects of NAMPT treatment on the SCN clock activity in SCN organotypic slice cultures obtained from PER2:LUC mice. SCN clock activity was determined by bioluminescence monitoring of PER2:luciferase activity. Representative circadian fluctuation curve are presented. The average values of period and amplitude of fluctuations were compared and plotted (n = 4 brain slices). One brain slice was obtained from one mice. Unpaired t-test (two-sided). n.s.: not significant. b Effects of intracerebroventricular (i.c.v.) administration of NAMPT protein (100 ng) on hypothalamic mRNA levels of genes. NAMPT was administered at ZT5, which was 1 h before sacrifice (Clock, Pomc NAMPT, Ox2r, n = 4; other groups, n = 5 mice). Unpaired t-test (two-sided). n.s.: not significant. SCN: suprachiasmatic nucleus, ARC: arcuate nucleus, DMN: dorsomedial nucleus. c Inhibition of the NAMPT (100 ng)-induced changes in hypothalamic Pomc, Npy, and Agrp mRNA levels by FK866 (100 ng) pretreatment (n = 4 mice). One-way ANOVA followed by Fisher’s LSD test. d, e POMC and NPY/AgRP neuronal activities measured with c-Fos expression at ZT6 and ZT18 after i.p. administration of saline or NAMPT (1 mg/kg) at ZT5 or saline or FK866 (10 mg/kg) at ZT17 (NPY c-Fos FK866, n = 4; other groups, n = 3 mice). The percentages of c-Fos⁺ POMC and NPY/AgRP neurons are presented with representative staining images. Unpaired t-test (two-sided) at ZT6 or ZT18. Scale bar = 50 μm. 3 V: third cerebroventricle. f Effects of i.c.v. administration of NAMPT (100 ng) alone or with SHU9119 (1 μg) on LMA and EE (Saline and NAMPT + SHU9119, n = 5; NAMPT alone, n = 4 mice). One-way ANOVA followed by Fisher’s LSD test. Two independent replicates were performed for all studies. Data are presented as mean ± SEM.