Slc17a6 cluster 3 differentially expressed Tac1 (gene encoding substance P), suggesting that these cells may be parapyramidal raphe neurons, whereas cluster four differentially expressed tyrosine hydroxylase (Th) indicative of adrenergic C1 pre-sympathetic neurons. Slc17a6 clusters 1–2 are presumed to be subsets of RTN chemoreceptors based on expression of Phox2b, Nmb, Gpr4, and Kcnk5 that differ in expression of galanin.
Cluster number is noted on the x axis and gene expression (from 0 to 4 counts/cell) on the y axis. ( B) UMAP plot depicting four sub-clusters of glutamatergic neurons and corresponding violin plots showing cluster-specific differential gene expression. A fairly large population of Chat expressing neurons was also detected but since neither glutamatergic ( B) nor Vgat+ neurons express Chat, this population was not analyzed further. Cells expressing either Slc17a6 or Slc32a1 were used for sub-cluster analysis of glutamatergic and inhibitory neurons. Neurons were differentiated from non-neurons (gray) based on expression of Snap25, Syp, Tubb3, and Elavl2. ( A) Through a normalized dispersion analysis for dimension reduction, a t-distributed stochastic neighbor embedding (t-SNE) was created of ventral parafacial single-cell transcriptome, with cells color coded by cluster.
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