Project description:Purpose: To characterize the estrogen receptor α (ESR1)-dependent transcriptome of neurons in the arcuate nucleus that regulate female bone density
Project description:From the arcuate nucleus of mice, we have dissociated neurons that express Proopiomelanocortin(POMC) and captured them on the Fluidigm C1 platform. These neuronsplay important roles in the regulation of appetite and sexual behavior. Here we propose tosequence RNA from 96 single POMC-expressing neurons to understand the variance in theirtranscriptomes.This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description: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.
Project description:We report RNA sequencing of single Agrp neurons isolated from the arcuate nucleus of the hypothalamus. Analyses of Agrp neuron transcriptomes reveals differential expression of receptors for multiple neuromodulators. Neuroanatomical mapping of known ligands of the receptors define subsets of neurons directly upstream of AgRP neurons in specific brain areas.
Project description:Drop-seq and single cell sequencing of mouse arcuate nucleus and median eminence. Please see below link for searchable cluster-based gene expression.
Project description:Background: Brain glucose-sensing neurons detect glucose fluctuations and prevent severe hypoglycemia, but mechanisms mediating functions of these glucose-sensing neurons are unclear. Methods: We combined mouse genetics, electrophysiology, neural tracing, optogenetics and Patch-RNA-seq to demonstrate how glucose-sensing neurons in the ventrolateral VMH regulate glucose balance. Results: Here we report that estrogen receptor-α (ERα)-expressing neurons in the ventrolateral subdivision of the ventromedial hypothalamic nucleus (vlVMH) are glucose-sensing neurons to a 5-1-5mM glucose fluctuation, being glucose-inhibited neurons (GI-ERαvlVMH) or glucose-excited neurons (GE-ERαvlVMH). Hypoglycemia activates GI-ERαvlVMH neurons via the anoctamin 4 channel, and inhibits GE-ERαvlVMH neurons through opening the ATP-sensitive potassium channel. Further, we show that GI-ERαvlVMH neurons preferentially project to the medioposterior arcuate nucleus of the hypothalamus (mpARH) and GE-ERαvlVMH neurons preferentially project to the dorsal Raphe nuclei (DRN). Activation of ERαvlVMH-mpARH circuit and inhibition of ERαvlVMH-DRN circuit both increase blood glucose. Conclusions: Our results indicate that ERαvlVMH neurons detect glucose fluctuations and prevent severe hypoglycemia in mice.
Project description:Drop-Seq single cell RNA sequencing of hypothalamic arcuate nucleus and median eminence from wildtype and Irx3/5 double heterozygous mice
Project description:We aimed to identify genes enriched in Nkx2-1-positive neurons in the dorsomedial hypothalamus (DMH). Using Nkx2-1-CreERT2 mice crossed with Rosa-ZsGreen mice after tamoxifen induction, we collected ZsGreen-marked Nkx2-1-positive neurons from the DMH, ventromedial hypothalamic nucleus (VMH), and arcuate nucleus (Arc) by laser microdissection and compared their gene expression profiles.
