Project description:We create catalogues of genes showing significant strain, parent-of-origin, dominance, sex effect in inbreds and reciprocal F1 hybrids of three wild-derived strains (CAST, PWK, WSB) across 4 different tissues (brain, kidney, liver, and lung) We used microarrays to validate the Brain results of RNAseq from inbred and F1 of 3 by 3 diallel. Brain, liver, kidney and lung RNA from the same mice used for RNAseq were hybridized to Affymetrix Mouse Gene 1.1 ST 96-Array Plate arrays using a GeneTitan instrument from Affymetrix.
Project description:To elucidate the function of Nova1 in neurons, especially in inhibitory neurons, gene expression profiling analysis (RNAseq, TRAP-seq) was performed using Nova1 conditional knockout mice and their primary neurons.
Project description:RNAseq of Adult Brain in ADAR1 E861A, ADAR2 knock out, ADAR1 E861A x ADAR2 knock out het, and ADAR1 E861A x ADAR2 knock out mice generated by deep sequencing.
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:Proteolytic ectodomain shedding of membrane proteins is a fundamental mechanism to control the communication between cells and their environment. A key protease for membrane protein shedding is ADAM17, which requires a non-proteolytic subunit, either inactive Rhomboid 1 (iRhom1) or iRhom2 for its activity. While iRhom1 and iRhom2 are coexpressed in most tissues and appear to have largely redundant functions, the brain is an organ with predominant expression of iRhom1. Yet, little is known about the spatio-temporal expression of iRhom1 in mammalian brain and about its function in controlling membrane protein shedding in the nervous system. Here, we demonstrate that iRhom1 is expressed in mouse brain from the prenatal stage to adulthood with a peak in early postnatal development. In the adult mouse brain iRhom1 was widely expressed, including in cortex, hippocampus, olfactory bulb and cerebellum. Proteomic analysis of the secretome of primary neurons and of cerebrospinal fluid (CSF), obtained from iRhom1-deficient and control mice, identified several membrane proteins that require iRhom1 for their shedding in vitro or in vivo. One of these proteins was the ‘multiple-EGF-like-domains protein 10’ (MEGF10), a phagocytic receptor in the brain that is linked to the removal of amyloid and apoptotic neurons. MEGF10 was further validated as an ADAM17 substrate using ADAM17-deficient mouse embryonic fibroblasts. Taken together, this study discovers a role for iRhom1 in controlling membrane protein shedding in the mouse brain, establishes MEGF10 as an iRhom1-dependent ADAM17 substrate and demonstrates that iRhom1 is widely expressed in murine brain.
Project description:We have performed microarray expression profiling of mouse primary neurons (cortical neurons and granule cell neurons) to model molecular networks and define whether distinct antiviral IFN responses occurred in neurons corresponding to different brain regions. Primary mouse cortical neurons and granule cell neurons were left untreated, treated with 100IU/mL of IFN-M-NM-2, or infected with West Nile virus (MOI of 1) and RNA was harvested after 24 hours. Three independent experiments were performed using a balanced design.
Project description:TRAP translational profiling is a method that allows investigators to genetically characterize specific cell types in complex tissues such as mouse brain. Using this technique we obtained RNA-Seq data from actively translating transcripts present in neurons in the hypothalamus of adult Lhx5-EGFP/Rpl10a (DT154) mice.
Project description:TRAP translational profiling is a method that allows investigators to genetically characterize specific cell types in complex tissues such as mouse brain. Using this technique we obtained RNA-Seq data from actively translating transcripts present in neurons in the thalamus of adult Stard8-EGFP/Rpl10a (ES2342) mice.