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. 2024 Apr 24;15:3443. doi: 10.1038/s41467-024-47877-2

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — A Reporter mice were treated with tamoxifen at 12 weeks of age and exposed to HFD, CD or (HFD-CD switch) for different durations 2 weeks later (t0). B, C Quantification of Pomc+ and Ghost neurons in Pomc^CreERT2;ZsGreen male mice fed with CD or HFD (6 months). Pomc mRNA was analyzed by FISH (B, n = 8 per group) or smFISH (C, n = 6 per group). D Quantification by IHC of Pomc+ and Ghost neurons in Pomc^CreERT2;ZsGreen male mice fed with CD (n = 4), HFD (7 months, n = 4), or exposed to HFD-CD switch (n = 3). E UMAP representation of 76 neurons analyzed by patch-seq in Pomc^CreERT2;tdTomato male mice exposed to CD (n = 5) or HFD (n = 5) for 6 months. We collected n = 38 cells from CD mice and n = 38 from HFD mice. We identified 4 main clusters: a (45 cells), b (13 cells), c (12 cells) and d (6 cells). F Heatmap of the relative gene expression (mean expression one cluster relative to the others) of Typical Pomc+ markers altered in Ghost neurons based on the data in Fig. 1D. G Heatmap of the relative gene expression (mean expression of one cluster relative to the others) of Pomc-neuron specific genes from HypoMAP in the clusters identified in (E). H Change in the % of cells in each cluster between the CD and HFD conditions. I Effects of HFD exposure (6 months) on electrophysiological parameters in the different clusters that were significantly different (HFD vs. CD) by two-tailed t test. RMP resting membrane potential. AP action potential. fAHP fast afterhyperpolarization. mAHP medium afterhyperpolarization. EPSCs: excitatory post synaptic currents. HFD high fat diet, CD chow diet. Data in (B−D) are mean ± s.e.m from 3 (B, C) or 2 (D) independent experiments. Data in (E) were obtained from 3 independent experiments. n indicates the individual biological values. In (B, C) *P = 0.0207 and **P = 0.0087 by two-tailed t test (CD Ghost vs HFD Ghost). In (D), **P = 0.0058 (CD Ghost vs HFD Ghost) and **P = 0.0022 (HFD Ghost vs HFD-CD Ghost) by ANOVA followed by Tukey’s post-test. Data in (H) are shown as a percentage of the total population P = 0,0004 by chi-squared test. Source data are in the Source Data file.