Single-cell analysis reveals cellular heterogeneity and molecular determinants of hypothalamic leptin-receptor cells

Hypothalamic nuclei which regulate homeostatic functions express leptin receptor (LepR), the primary target of the satiety hormone leptin. Single-cell RNA sequencing (scRNA-seq) has facilitated the discovery of a variety of hypothalamic cell types. However, low abundance of LepR transcripts prevented further characterization of LepR cells. Therefore, we perform scRNA-seq on isolated LepR cells and identify eight neuronal clusters, including three uncharacterized Trh-expressing populations as well as 17 non-neuronal populations including tanycytes, oligodendrocytes and endothelial cells. This includes food restriction to measure the response of Lepr positive cells to fasting. Using FACS sorting into 384 well plates and Celseq2 method, we generated a high-quality dataset containing a median of 12,082 unique counts and 4,756 genes per cell. Multiplexed (384) single cell libraries of sorted Lepr cells of the hypothalamus of Lepr-cre;Rosa-LSL-tdTomato animals, library 1 Multiplexed (384) single cell libraries of sorted Lepr cells of the hypothalamus of Lepr-cre;Rosa-LSL-tdTomato animals, library 2 Multiplexed (384) single cell libraries of sorted Lepr cells of the hypothalamus of Lepr-cre;Rosa-LSL-tdTomato animals, library 3 Multiplexed (384) single cell libraries of sorted Lepr cells of the hypothalamus of Lepr-cre;Rosa-LSL-tdTomato animals following 24 of fasting, library 4