Extended Data Fig. 7: Inflammation drives ECM remodeling within the ARC to promote metabolic dysfunction.
From: Pathogenic hypothalamic extracellular matrix promotes metabolic disease
a) C57BL/6 J mice were fed a HFHS diet for 12 weeks and brains were extracted for assessment of pro-fibrotic gene expression. b) 10-week-old C57BL/6 J mice received bilateral intraARC injections of anti-inflammatory AAV vectors expressing soluble receptors for TNFR1α and TGFβR or control, before being placed on a HFHS for 8-weeks. Brains were extracted and processed for c,d) ARC WFA immunofluorescence and e) CS-GAG abundance. f) Body weight, g) fasting induced 2-hour food intake, h) energy expenditure and i) glycemic control were assessed. j) 10-week-old C57BL/6 J mice received bilateral injections into the ARC to administer inflammatory AAV vectors expressing TNFα and TGFβ ligands, or control. 6-weeks later mice received bilateral intraARC injections of vehicle or chABC to enzymatically digest the ARC PNN and k,l) brains were processed for WFA immunostaining or m) quantitative analysis of CS-GAG abundance. n) Body weight, o) adiposity, p) fasting induced 2-hour food intake, q) energy expenditure and r) glycemic control was assessed. Results are mean ± SEM; significance determined using o, p, q, r) one-way ANOVA with Tukey’s multiple comparisons, f, n) two-way ANOVA with multiple comparisons, a, d, e, g, h, i, m) two-tailed t-tests. n = biologically independent samples and significance is representative of two independent experiments. Scale bar, 100 µm. Abbreviations: III, 3rd ventricle; ARC, arcuate nucleus of the hypothalamus; AUC, area under the curve; BW, body weight; chABC, chondroitinase ABC; CS-GAG, chondroitin sulfate glycosaminoglycans; GFP, green fluorescent protein; HA, hyaluronan acid; HFHS, high-fat high-sugar diet; IF, inflammatory; WFA, Wisteria Floribunda Lectin.
