Fig. 1: Lineage tracing of Pomc neurons in adult chow-fed mice reveals subtypes with atypical molecular identity. | Nature Communications

Fig. 1: Lineage tracing of Pomc neurons in adult chow-fed mice reveals subtypes with atypical molecular identity.

From: Single cell tracing of Pomc neurons reveals recruitment of ‘Ghost’ subtypes with atypical identity in a mouse model of obesity

Fig. 1

A Detection and quantification of Pomc+ and Ghost neurons in PomcCreERT2;ZsGreen male mice by analyzing reporter cells (ZsGreen) positive (Pomc + ) or negative (Ghost) for Pomc mRNA (FISH, n = 11 mice) or Pomc protein (IHC, n = 5 mice). B Detection and quantification of Pomc+ and Ghost neurons in PomcCreERT2;tdTomato male mice by analyzing reporter cells (tdTomato) positive (Pomc + ) or negative (Ghost) for Pomc mRNA (FISH, n = 3 mice) or Pomc protein (IHC, n = 4 mice). C Detection and quantification of Pomc+ and Ghost neurons in PomcCreERT2;tdTomato;Pomc-eGFP male mice (n = 4) by analyzing reporter cells (tdTomato) positive (Pomc + ) or negative (Ghost) for endogenous eGFP protein by IHC. D Quantification of Pomc+ and Ghost neurons co-expressing detectable mRNA or protein levels of several Pomc neuronal functional markers in PomcCreERT2;ZsGreen or PomcCreERT2;tdTomato male mice. Cart mRNA was assessed by smFISH (n = 5 mice), Tbx3 by IHC (n = 11 mice), hormone receptors by smFISH (n = 4 mice), and neurotransmitter-related markers by smFISH (n = 6 mice). E Pomc neurons in the hypothalamus by the scRNAseq atlas HypoMap. Left: Global Uniform Manifold Approximation and Projection (UMAP) with Pomc-cluster position highlighted. Middle: Summarised Pomc expression levels within the POMC-cluster. Right: Pomc neuron subclusters. F Representative image and quantification of the percentage of Pomc+ and Ghost neurons co-postive for Glipr1 mRNA by smFISH in PomcCreERT2;tdTomato male mice (n = 6). Pomc: pro-opiomelanocortin, AgRP: agouti-related protein, Npy: neuropeptide y, Kiss: kisspeptin, Cart: cocaine-and amphetamine-regulated transcript, Tbx3: T-box gene 3, Lepr: leptin receptor, Glp1r: glucagon-like peptide-1 receptors Gad1/2: Glutamate decarboxylase 1 and 2, Slc17a6: vesicular glutamate transporter 2, IHC: immunohistochemistry, FISH fluorescence in situ hybridisation, smFISH single molecule fluorescence in situ hybridisation, NT neurotransmitter. Glipr1: Glioma pathogenesis-related protein 1. Pomc+ neurons are indicated by arrowheads, Ghost neurons by arrows. Data in (A−D, and F) are mean ± s.e.m. from 3 independent experiments. n indicates the individual biological values. Data in (A−D and F) were obtained 2 weeks after adult onset (12-week-old mice) administration of tamoxifen. In D: ***P = 0.0004 (Cart), *P = 0.0453 (Tbx3), *P = 0.0129 (Lepr), *P = 0.049 (Glp1r), **P = 0.0117 (Insr),*P = 0.016 (Slc17a6) by two-tailed t test. In F, *P = 0.0154 by two-tailed t-test. Source data are provided as a Source Data file.

Back to article page