Figure 5

VGF is expressed in neural pathways that innervate brown adipose tissue (BAT). In panel A, levels of VGF-derived peptides were quantified in BAT from C57Bl6 female mice fed standard chow or a high fat, high carbohydrate diet for 14 days. BAT was removed and RIA analysis carried out to quantify C-terminal VGF-derived peptides (NAPP129, TLQP62 and AQEE30, but not full length VGF), using polyclonal anti-AQEE30 (VGF_588-617) antiserum as previously described [38]. VGF peptide levels in BAT were significantly reduced in mice fed the high-energy diet in comparison to control mice (p < 0.05). Comparisons by ANOVA [mean ± SEM; high fat diet (n = 4), standard diet (n = 3)]. In panels B-C, Vgf gene expression was visualized in neural pathways that innervate tissues including BAT using a lacZ reporter gene that was knocked into the mouse Vgf locus in VGF_R mice (see Methods). In panel B, β-galactosidase expression (blue staining) in the dorsal root ganglia (white arrows) and intercostal nerve (black arrow) is seen in a whole mount preparation from a heterozygous Vgf _R +/- mouse. A dorsal root ganglion section is shown at higher magnification in panel C (scale bar is 100 μm in length).