Fig. 7: Validation of heterogeneous neuronal populations.
From: HypoMap—a unified single-cell gene expression atlas of the murine hypothalamus
a–c, Glp1r-expressing cell types identified the hypothalamus. a, The Glp1rCre bacTRAP signature is enriched in multiple hypothalamic cell types, mostly corresponding to the Glp1r expression in HypoMap. b,c, RNAscope of Glp1r together with specific markers of neuron clusters identified using Glp1r-bacTRAP in (a). Representative images (b) and quantification shown as the percentage Glp1r-positive cells identified by marker gene expression (c). Points refer to individual sections, in total 4 rostral and 4 caudal ARC sections from 4 mice were included for each experiment (0 rostral and 8 caudal for Tbx19- plus Anxa2 and 16 PVH sections for Oxt). Mean ± s.e.m,: Pomc: 49.03 ± 4.77; Pomc/Anxa2: 14.56 ± 3.89; Sst: 31.39 ± 3.06; Sst/Unc13c: 64.46 ± 3.88; Ghrh: 47.93 ± 5.19; Tbx19/Anxa2: 38.13 ± 8.3; Trh/Nkx2-4: 90.89 ± 8.26; Oxt: 47.94 ± 6.73. d–f, Pnoc-expressing cell types identified in the hypothalamus. d, The PnocCre bacTRAP signature is enriched in multiple hypothalamic cell types, and covers only a subset of Pnoc-expressing cell types in HypoMap. e,f, RNAscope of Pnoc and marker genes of selected ARC neuronal cell types based on Pnoc-bacTRAP and gene expression in (d). e,f, Representative images (e) and quantification (f) of Pnoc and Sst or Crabp1 co-expressing subclusters. Points refer to individual sections, in total 14 ARC sections along the rostral-caudal axis from 4 mice were included for each experiment. Mean ± s.e.m: Sst/Pnoc: 59.9 ± 3.24; Sst/Pnoc/Nts: 6.78 ± 3.56; Sst/Pnoc/Unc13c: 52.28 ± 7.17; Sst/Pnoc/Nts/Unc13: 2.29 ± 1.1; Crabp1/Pnoc: 76.35 ± 2.2; Crabp1/Pnoc/Tmem215: 41.02 ± 4.81; Crabp1/Pnoc/Htr3b: 32.18 ± 3.04; Crabp1/Pnoc/Tmem215/Htr3b: 10.02 ± 1.17. In all dot plots, the red point depicts the mean and red error bars the s.e.m. of all sections. We used a two-sided Wilcoxon rank-sum test (multiple testing correction with Benjamini–Hochberg) to test for differences between the means of relevant groups and added the resulting P values to the quantification in (c) and (f).
