Supplementary Figure 4: Neuropeptide-defined subtypes of ARCVglut2 neurons and ARCVglut2 single-neuron gene expression of synapse-related genes.
From: A rapidly acting glutamatergic ARC→PVH satiety circuit postsynaptically regulated by α-MSH
a, Schematic of single-cell RNA-seq clustering analysis based on expression of 90 neuropeptide genes (see Supplemental Table 1). b, Two-dimensional representation of ARCVglut2 neurons (dots) by t-distributed stochastic neighbor embedding (tSNE). Subtypes are annotated according to neuropeptide gene markers. One subtype was not specifically marked by expression of a single neuropeptide (or other) gene and is annotated as “Other.” c, tSNE plot re-colored to show expression of prominent neuropeptide marker genes. Colors of dots indicate the level of expression relative to the maximal level for each gene. c, Expression of synapse-related genes in Pomc-positive (Pomc+) versus Pomc-negative (Pomc-) subsets of ARCVglut2 neurons. Sample size (cells): Pomc-positive/Pomc-negative, n = 10/13. Slc17a6: Pomc-positive (mean = 21.63, s.e.m. = 5.38) versus Pomc-negative (mean = 62.86, s.e.m. = 11.2): Unpaired two-tailed t-test; t(21) = 3.016, **P = 0.0066. Syt1: Pomc-positive (mean = 0.37, s.e.m. = 0.209) versus Pomc-negative (mean = 1.23, s.e.m. = 0.2662): Two-tailed Mann-Whitney test; U = 21, **P = 0.0052. Cplx1: Pomc-positive (mean = 3.412, s.e.m. = 2.054) versus Pomc-negative (mean = 20.26, s.e.m. = 5.574): Two-tailed Mann-Whitney test; U = 20, **P = 0.0041. Data are presented as mean ± s.e.m.
