Project description:We report phosphoRiboTrap experiments result which aim to explore the nature of the cell types in the Median Eminence (ME) regulated as a consequene of chemogenetic MCH neuron activation. Control and MCH-hM3Dq mice were 12 hrs-fasted and i.p. injected with CNO (3 mg/kg), Arcuate (ARC) and median eminence expat were extracted for preciptation of S6-marked ribosomes from both groups of mice. Extracted RNA from Immunoprecipitated ribosomes (IP) and total tissue (ARC+ME) from each mouse were subjected to deep mRNA sequencing. By analyzing the overlap of genes enriched in the IP/Input of MCH neuron activated mice with previously identified cell types using single cell mRNA sequencing of cells in the mediobasal hypothalamus (Campbell et al., 2017), we identified gene clusters which were activated upon MCH neuron activation.
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: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:Single cell sequencing of Ins2-Cre;ptdTomato+ FACS sorted cells from micro-dissected hypothalamic arcuate nucleus and median eminence of wildtype and Irx3/5 double heterozygous mice.
Project description:The median eminence (ME) of the hypothalamus is a structure that rapidly adapts to nutrient availability. We have used single-cell RNA sequencing to characterize the transcriptional profiles of cells specifically in the median eminence in the fasted and refed states. In our study, we focus on characterizing the oligodendrocyte population of the ME, and identifying how subpopulations respond to nutrient availability.
Project description:Here we aim to decipher the actions of Irx5 in the regulation of obesity and metabolic abnormalities. We employed a mouse model homozygous for an Irx5-knockout (Irx5KO) allele and conducted droplet based single-cell RNA sequencing (Drop-seq) in the hypothalamic arcuate-median eminence (ARC-ME), which is the main control center for sensing and integrating feeding regulatory signals.
Project description:The hypothalamus is a functionally and cellularly complex tissue controlling many developmental processes, including puberty. While key hormonal aspects of puberty regulation in the hypothalamus are well established, understanding the genes, cell-types, and epigenetic mechanisms underlying and regulating puberty is limited. Here, we performed 3’-UTR-seq on the hypothalamus from both sexes of C57BL/6J mice at 5 ages spanning pubertal transition (postnatal days 12, 22, 27, 32, 37) (4-5 replicates per sex at each age) to examine genome-wide age- and sex-biased trends in gene expression in a cell-type aware manner. Sample collection, RNA extraction, and sequencing was completed using the same protocols and on the same mice as PMID: 3622112 (E-MTAB-9459). QuantSeq 3’mRNA-seq libraries were constructed from total RNA using an automated method with Agilent NGS Workstation. The resulting single-end libraries were sequenced at SickKids TCAG core on the Illumina v4 flow cell with SR50 bp cycles extended to 68 bp. A customized pipeline was developed and used for the analysis of reads obtained (PMID: 36221127 for details). Processed reads were mapped to mouse genome (mm10).
Project description:Brain-derived serotonin favors appetite in mice following its binding to the Htr1a and Htr2b receptors in arcuate neurons of the hypothalamus. In this study, we identified that CREB is the transcriptional effector of brain-derived serotonin control of appetite in arcuate nuclei. In this dataset, we identified the downstream genes of CREB in arcuate neurons of the hypothalamus controling appetite. We isolated hypothalami of wild type and Creb-pomcCre-/- (deleted for Creb selectively in arcuate neurons of the hypothalamus) mice and performed microarray experiments.
