Fig. 1: Distribution of Ox1R and Ox2R in serotonergic neurons of the raphe nucleus of a SERTtdTomato mouse. | Nature Communications

Fig. 1: Distribution of Ox1R and Ox2R in serotonergic neurons of the raphe nucleus of a SERTtdTomato mouse.

From: Orexin receptors 1 and 2 in serotonergic neurons differentially regulate peripheral glucose metabolism in obesity

Fig. 1

a Representative images of RNAscope in situ hybridization in the dorsal raphe nucleus (dorsal (DRD), ventral (DRV)), and median raphe nucleus (dorsal (MRD), ventral (MRV)). b Representative images of RNAscope in situ hybridization in the raphe pallidus (RPa). c, e Percentages of Ox1R or Ox2R positive neurons in serotonergic neurons. d, f The average of relative fluorescence intensity of Ox1R or Ox2R signal in each serotonergic neuron. Fluorescence was normalized by the mean of all regions for each mouse. Blue, tdTomato; red, Ox1R; cyan, Ox2R. Scale bar: 100 µm in (a) and 50 µm in (b). n = 3. Data are represented as means ± SEM. d DRV: p = 0.035, MRD: p = 0.036; e DRD: p = 0.049, MRD: p = 0.0081, RPa: p = 0.028; f DRD: p = 0.010, MRV: p = 0.0008, RPa: p = 0.0099. *p < 0.05, **p < 0.01, ***p < 0.001; compared to DRD (d) or DRV (e, f), as determined by paired two-tailed t-test (c, e) or ratio paired two-tailed t-test (d, f). Analysis of published scRNA-Seq data of serotonergic neurons in dorsal raphe nucleus (DR)30,31: g Scatter plot of raw cell read counts of Ox1R and Ox2 R, g’ percentages of serotonergic neurons only expressing Ox1R (Ox1R-only) or Ox2R (Ox2R-only) and those expressing both or none of Ox1R and Ox2R (Ox1/2R+ or Ox1/2 R−), h scatter plot of raw vesicular glutamate transporter 3 (VGLUT3) counts against raw Ox1R or Ox2R counts, h’ percentages of VGLUT3 positive (+) and negative (−) neurons in Ox1R-only and h” Ox2R-only serotonergic neurons, and i heat map of gene counts for cells with column-wise z-scores. Mean gene abundance was calculated per gene (x-dimension) and group (y-dimension). Column-wise z-scoring (per gene) was applied to make expression visually comparable between groups independent of average gene abundance levels. j Representative images of RNAscope in situ hybridization in the dorsal raphe nucleus (scale bar: 100 µm), and analysis: k percentages of Ox1R-only, Ox2R-only, Ox1/2R+ and Ox1/2R− serotonergic neurons; percentages of VGLUT3 positive neurons in (l) Ox1R-only and (m) Ox2R-only serotonergic neurons. Gray, tryptophan hydroxylase isoform 2 (TPH2); red, Ox1R; cyan, Ox2R; blue, VGLUT3. n = 3. Source data are provided as a Source Data file.

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