Figure 4. Characterisation of Gipr^EYFP+ and Glp1r^EYFP+ neurons.

A) Neurons were subsetted based on positive expression of Syt1 and Snap25. Cells expressing high levels of Mustn1, Acta2, or Mal were excluded, yielding 1341 Gipr^EYFP+ and Glp1r^EYFP+ neurons for downstream analysis. B) UMAP showing 12 separate clusters of Gipr^EYFP+ and Glp1r^EYFP+ neurons following dimensionality reduction and unsupervised clustering. Cluster-specific markers were identified using negative binomial regression analysis (see Table 1). The top 15 cluster-specific markers were cross-referenced with published brain region-specific transcriptional markers and the Allan Brain Atlas for region-specific mapping of clusters (see Supplementary Fig 5). Percentages of neurons from either Gipr^EYFP+ or Glp1r^EYFP+ neurons for each cluster are represented in pie charts. Total cell number per cluster is expressed at the top of each pie chart. C. Violin plots of neurotransmitters, secreted products, and cell surface receptors enriched in each neuronal cluster as determined by negative binomial regression analysis. Data are plotted in counts per million (CPM). ARH: arcuate hypothalamic nucleus, VMH: ventromedial hypothalamic nucleus, SCN: suprachiasmatic nucleus, LH: lateral hypothalamic nucleus, PM: premammillary nucleus, MTu: medial tuberal nucleus.