The transcriptomics of SLC18A3 positive cells from arcuate nucleus of rats in different pubertal development stages.

Based on our spatial transcriptomics sequencing data, we observed solute carrier family 18 member A3 (SLC18A3) was specificially expressed in kisspeptin-expressed neurons in arcuate nucleus (ARC). SLC18A3 as a surface marker was used to examine the isolation of living kisspeptin-expressed neurons by flow cytometric (FACS) assay. Fresh isolated ARC nucleus tissues from female Sprague-Dawley rats in postnatal day (PND)-25, 35 and 45 (n=10 each group) were harvested, and processed single-cell suspension for FACS assay by SLC18A3. Here, we obtained two populations according to the cell size by FSC/SSC signals, and further harvested a small proportion of Slc18a3+ cells from hypothalamus of PND-25, 35 and 45. Smart-seq was conducted to detect the transcriptome of four population namely large Slc18a3+/- cells as well as small Slc18a3+/- cells.