Project description:VMH was dissected from 16-week-old mice with deletion of SIRT1 in SF1 neurons (Sf1-Cre; Sirt1loxP/loxP) and intact controls (Sirt1loxP/loxP) and fed on high calorie diet from 8 weeks of age.
Project description:Microdissected ventromedial hypothalamus (VMH) was profiled to identify transcriptional changes after Nkx2-1 ablation in female mice. Total RNA was extracted from conditional mutants (Nkx2-1 f/f; Sf1Cre) and floxed controls (Nkx2-1 f/f) at postnatal day 10 (P10) and profiled using Mouse ref8 v2.0 Illumina chips and reagents (n=4/group). The tissue profiled is microdissected ventromedial hypothalamus from P10 female mice. Controls and mutants are analyzed in quadruplicate.
Project description:To uncover the cellular architecture of the mouse ventromedial hypothalamus (VMH), we used single nucleus RNA-seq (snRNA-seq) from wild-type mice.
Project description:Analysis of gene expression regulated by FoxO1 in the ventromedial hypothalamus (VMH) of wildtype and knockout mice. Results provide important information of gene expression in the VMH. The transcription factor FoxO1 contributes to leptin and insulin action, including cells in the brain. However, the neurons mediating these effects and the identity of the molecular targets through which FoxO1 exerts metabolic actions remains to be defined.
Project description:Loss of synchrony between geophysical time and insulin action predisposes to metabolic diseases. Yet, the brain and peripheral pathways linking proper insulin effect to diurnal changes in light/dark and feeding/fasting inputs are poorly understood. Here, we show that insulin sensitivity of several metabolically relevant tissues fluctuates during the 24-hour period. For example, in mice insulin sensitivity of skeletal muscle, liver, and adipose tissue is lowest during the light period. Mechanistically, by performing loss- and gain-of-light-action and food-restriction experiments, we demonstrate that SIRT1 in steroidogenic factor 1 (SF1) neurons of the ventromedial hypothalamic nucleus (VMH) conveys photic inputs toentrain the biochemical and metabolic action of insulin in skeletal muscle. These findings uncover a critical light-SF1-neuron-skeletal-muscle axis that acts to finely tune diurnal changes in insulin sensitivity and reveal a light regulatory mechanism of skeletal muscle function.
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.
Project description:Analysis of gene expression regulated by FoxO1 in the ventromedial hypothalamus (VMH) of wildtype and knockout mice. Results provide important information of gene expression in the VMH. The transcription factor FoxO1 contributes to leptin and insulin action, including cells in the brain. However, the neurons mediating these effects and the identity of the molecular targets through which FoxO1 exerts metabolic actions remains to be defined. For the gene expression profiling, total RNA was obtained from the VMH of WT and VMH-specific FoxO1 KO mice using Qiagen RNeasy Lipid Tissue Mini Kit (Qiagen Sciences, Maryland) and Phase Lock Gel (5 Prime Inc., Gaithersburg, MD). To make sure reproducibility and biological significance, triple hybridizations were performed for each genotype with the RNA from three independent VMH samples, each sample contains RNA from three animals (biological replicates). Genomics and Microarray Core Facility at UT-Southwestern (http://microarray.swmed.edu/) checked RNA quality with Bioanalyzer Chip and processed the samples for hybridization with Illumina Mouse-6 V2 BeadChip (Illumina Inc., San Diego, CA). We used Partek Genomics Suite 6.5 (Partek Inc., St Louis, MO) and Ingenuity Pathway Analysis (Ingenuity Systems, Inc., Redwood City, CA) for data analysis.
Project description:To uncover the cellular architecture of the macaque ventromedial (VMH) and dorsomedial hypothalamus (DMH), we used single nucleus RNA-seq (snRNA-seq) from a Rhesus macaque
