Fig. 3: Identification of CCN3 as a brain-derived osteoanabolic factor.
a, Trabecular and cortical fractional BV, mechanical strength (three-point bend) and BMAT levels in long bones of Esr1fl/fl and Esr1Nkx2.1-cre females fed standard diet (SD) or HFD for 17 weeks (N = 4–6 per group). b, Representative images of tibia from female mice (aged 27 weeks) fed SD or HFD for 17 weeks labelled for calcein and Alizarin Red (top, white arrows and magnified from Extended Data Fig. 6c) and osmium stained with lipid droplets (bottom, yellow arrows). c, Heatmap of top DEGs changed in the ARC of Esr1Nkx2.1-cre females at 12 weeks of age (adapted from ref. 2) and at 27 weeks of age fed SD or HFD. Scale is log2 fold change. d, Transcript levels of Ccn3 and Penk in the ARC of 3.5-week-old mutant females, measured by quantitative PCR (qPCR). N = 2–3 per group. e, Ccn3 and Penk expression by RNAscope of the ARC in mutant female Esr1Nkx2.1cre mice fed either SD or HFD. Scale bar, 100 µm. ME, median eminence. f, Staining for ERα (pink) and CCN3 (green) in brain sections from posterior ARC and SCN regions of Esr1fl/fl female (10-week-old) and Esr1Nkx2.1-cre female and male (12-week-old) mice. Scale bar, 200 µm. oc, optic chiasm. g, CCN3 and KISS1 overlapping expression in Esr1Nkx2.1-cre female medial basal posterior ARC (yellow arrowheads). Scale bar, 100 µm. h, Ccn3 (green) Esr1 (cyan) and Kiss1 (red) transcripts from posterior ARC brain sections in control and mutant Esr1Nkx2.1-cre females (Kiss1 only, yellow arrowheads; Ccn3 only, white arrowheads). Scale bar, 50 µm. One-way ANOVA in a (Tukey’s and Šidák’s multiple-comparisons test). Unpaired Student’s t-test, two-tailed for d. **P < 0.01, ***P < 0.001, ****P < 0.0001. Error bars ± s.e.m.
