Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Validation of molecularly-defined olfactory bulb projection neurons targeting anterior or posterior brain areas

ABSTRACT: To validate different projection targets of already molecularly-defined olfactory bulb projection neurons we used viral targeting specifically into anterior or posterior cortical areas, Fluorescence Activated Nuclei Sorting (FANS) to enrich for olfactory bulb projection neurons, and single-nuclei RNA sequencing (sn-RNA seq) To isolate GFP-labelled nuclei, 1 individual replicate of AON or PCx-injected mice was used. Ipsilateral and controlateral sides were minced separately and placed into two different tubes. The minced tissue was gently homogenized in Nuclei PURE Lysis Buffer and 10% Triton X-100 using an ice-cold dounce and pestle, and filtered two times through a 40 μm cell strainer on ice. After centrifuging at 500 rpm for 5 min at 4 °C, the supernatant was aspirated and gently resuspended in 500 μl of cold buffer (1x of cold Hanks' Balanced Salt Solution HBSS, 1% nuclease-free BSA, RNasin Plus and 1/2000 DRAQ5). Our study identifies molecularly distinct subtypes of mitral cells projecting to anterior or posterior olfactory cortices.

ORGANISM(S): Mus musculus

PROVIDER: GSE176350 | GEO | 2021/06/09

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

Molecular diversity of olfactory bulb projection neurons [single-cell RNA-seq]

Project description:To characterize the molecular diversity of olfactory bulb projection neurons we used viral targeting and Fluorescence Activated Nuclei Sorting (FANS) to enrich for olfactory bulb projection neurons, and single-nuclei RNA sequencing (sn-RNA seq) to comprehensively characterize their transcriptomes. To isolate GFP-labelled nuclei, 3 individual replicates of AON and PCx-injected mice were used. Ipsilateral and controlateral sides were minced separately and placed into two different tubes. The minced tissue was gently homogenized in Nuclei PURE Lysis Buffer and 10% Triton X-100 using an ice-cold dounce and pestle, and filtered two times through a 40 μm cell strainer on ice. After centrifuging at 500 rpm for 5 min at 4 °C, the supernatant was aspirated and gently resuspended in 500 μl of cold buffer (1x of cold Hanks' Balanced Salt Solution HBSS, 1% nuclease-free BSA, RNasin Plus and 1/2000 DRAQ5). Our study identifies molecularly distinct subtypes of mitral and tufted cells.

2020-12-05 | GSE162654 | GEO

Retrograde labelling and RNA deep sequencing of olfactory bulb projection neurons [bulk RNA-seq]

2020-12-05 | GSE162655 | GEO

Snip1 and PRC2 coordinate the intrinsic apoptosis, cell division, and neurogenesis in the developing brain [CUT&RUN]

Project description:Approximately 3 × 105 NPCs sorted for Sox2-eGFP were mixed with 3 × 104 Drosophila S2 cells per reaction. We performed CUT&RUN using the protocol developed by the Henikoff lab (Skene and Henikoff, 2017). In brief, Bio-Mag®Plus Concanavalin-A (Con A) coated beads (Bangs Laboratories BP531) were washed and activated with Binding buffer. Nuclei of NPCs with S2 spike-in were gently prepared (see Subcellular Protein Extraction section). Activated Con A beads and nuclei were then mixed and rotated for 5 min at room temperature. They were then blocked with Digitonin block buffer for 5 min at room temperature. 0.5-1 μg of primary antibody with 0.25 μg Spike-in antibody (Active Motif 61686) diluted in Digitonin block buffer was added to the bead-nuclei mixture and incubated for 3 hours (histone marks) or O/N (the rest) at 4 °C. Beads were washed three times with Digitonin block buffer and incubated with pA-MNase for 1 hour at 4 °C. Beads were washed three times with Wash buffer and incubated in Wash buffer for 10 min on ice. The pA-MNase was activated by incubating the beads with 2 mM CaCl2 for 25 min on ice and quenched by adding the Stop buffer. DNA was released from the beads by incubating them for 30 min at 37 °C and collected by centrifugation at 16,000 x g for 5 min at 4 °C. DNA was then isolated by using a phenol/chloroform extraction method.

2023-07-12 | GSE212444 | GEO

Validation of molecularly-defined olfactory bulb projection neurons targeting anterior or posterior brain areas

Project description:Validation of molecularly-defined olfactory bulb projection neurons targeting anterior or posterior brain areas

| PRJNA735939 | ENA

Molecular diversity of olfactory bulb projection neurons

Project description:Molecular diversity of olfactory bulb projection neurons

| PRJNA682577 | ENA

ChIP-seq of bovine cells (MDBK) to study butyrate-induced histone modification

Project description:Utilizing next-generation sequencing technology, combined with ChIP (Chromatin Immunoprecipitation) technology to analyze histone modification (acetylation) induced by butyrate and to map the epigenomic landscape of normal histone H3, H4 Cells were treated with 10 mM butyrate for 24 hr, The cells were scraped from the flask and homogenized with ice-cold Dounce homogenizer to release the nuclei. The collected nuclei were resuspended in digestion buffer and enzymatic shearing was performed. ChIP with anti H3, H4 and acetyl-H3 and acetyl-H4.

2014-01-24 | E-GEOD-38973 | biostudies-arrayexpress

Real-time quantitative PCR array analysis of primary colon cancer and liver metastasis derived exosomes

Project description:To isolate exosomes from colon tissue, colon tissue were removed and gently disaggregated with tweezers in dissociation buffer, followed by incubation at 37°C for 1 h. The disaggregated samples were centrifuged at 1,000 g for 10 min, 2,000 g for 20 min, 4,000 g for 30 min and 10,000 g for 1 h with the supernatant being retained each time. The tissue exosomes were collected by centrifuging the samples at 100,000 g for 1.5 h at 4°C, and the pellet was suspended in ice-cold PBS. Total RNA was isolated from cells, exosomes and tissue using a miRNeasy mini kit (Qiagen). miRNA expression profiling including exosomes and their donor cells or tissues was performed using the Qiagen miScript miRNA PCR Array Mouse miRBase Profiler. To isolate exosomes from liver tissue, a 20-G catheter was inserted into the portal vein of anaesthetized mice. The perfused liver tissue, as well as colon tissue were removed and gently disaggregated with tweezers in dissociation buffer, followed by incubation at 37°C for 1 h. The disaggregated samples were centrifuged at 1,000 g for 10 min, 2,000 g for 20 min, 4,000 g for 30 min and 10,000 g for 1 h with the supernatant being retained each time. The tissue exosomes were collected by centrifuging the samples at 100,000 g for 1.5 h at 4°C, and the pellet was suspended in ice-cold PBS. Total RNA was isolated from cells, exosomes and tissue using a miRNeasy mini kit (Qiagen). miRNA expression profiling including exosomes and their donor cells or tissues was performed using the Qiagen miScript miRNA PCR Array Mouse miRBase Profiler.

2017-01-04 | GSE93092 | GEO

Single nuclei RNA-seq from adult mouse Hippocampus

Project description:We report RNA-seq of single nuclei isolated from the adult C57BL/6 male mouse Hippocampus region. Majority of the nuclei were isolated from 12 weeks old mice (4 different animal), with an additional set of nuclei from 3 months and 2 years old animals. In addition a set of GFP labeled nuclei driven by a VGAT promoter . Microdissections of dentate gyrus, CA1 and CA2/3 regions of the Hippocampus were placed into ice-cold RNA-later for fixation and stored at 4âc overnight, then stored in -80âc. Nuclei were isolated by sucrose gradient centrifugation and kept on ice until sorting using Fluorescence Activated Cell Sorting (FACS) into 96 well plates containing RNA lysis buffer. Single nucleus RNA was first purified then derived cDNA libraries were generated following a modified Smart-seq2 protocol. For VGAT nuclei: high titer AAV1/2 of pAAV-EF1a-DIO-EYFP-KASH-WPRE-hGH-polyA was injected into dorsal and/or ventral Hippocampus, animals were sacrificed two weeks after injections, and GFP labeled nuclei were sorted into plates and processed as described above.

2016-08-16 | E-GEOD-85721 | biostudies-arrayexpress

Transcriptional responses to deep cerebellar nuclei (DCN) and entorhinal cortex (EC) protein homogenates in cultured microglia

Project description:We hypothesised that enriched cytokines in the microenvironment of the deep cerebellar nuclei (DCN) contribute to the inflammatory gene expression profiles of DCN microglia and thus might induce a similar response in cultured microglia. To test this hypothesis, we treated BV2 microglia cell lines with DCN or entorhinal cortex (EC) protein homogenates and analysed RNA expression 4h post-stimulation, the peak timepoint of LPS-induced inflammatory gene expression (Das et al, 2016). To enrich for extracellular secreted proteins and to prevent effects of extraction buffer components on cell cultures, proteins were gently homogenised in DPBS on ice in the absence of detergent and protease inhibitors. As a positive control for inflammatory response gene induction, we concurrently treated BV2 cultures with LPS.

2023-08-31 | GSE223018 | GEO

Alpha-Synuclein pathology and reduced neurogenesis in the olfactory system affect olfaction in a mouse model of Parkinson's disease

Project description:Parkinson’s disease (PD) is characterized by multiple symptoms including olfactory dysfunction, whose underlying mechanisms remain unclear. Here, we explored pathologic changes in the olfactory pathway of transgenic (Tg) mice of both sexes expressing the human A30P mutant α-synuclein (α-syn; α-syn-Tg mice) at 6–7 and 12–14 months of age, representing early and late-stages of motor progression, respectively. α-Syn-Tg mice at late stages exhibited olfactory behavioral deficits, which correlated with severe α-syn pathology in projection neurons (PNs) of the olfactory pathway. In parallel, olfactory bulb (OB) neurogenesis in α-syn-Tg mice was reduced in the OB granule cells at six to seven months and OB periglomerular cells at 12–14 months, respectively, both of which could contribute to olfactory dysfunction. Proteomic analyses showed a disruption in endocytic and exocytic pathways in the OB during the early stages which appeared exacerbated at the synaptic terminals when the mice developed olfactory deficits at 12–14 months. Our data suggest that (1) the α-syn-Tg mice recapitulate the olfactory functional deficits seen in PD; (2) olfactory structures exhibit spatiotemporal disparities for vulnerability to α-syn pathology; (3) α-syn pathology is restricted to projection neurons in the olfactory pathway; (4) neurogenesis in adult α-syn-Tg mice is reduced in the OB; and (5) synaptic endocytosis and exocytosis defects in the OB may further explain olfactory deficits.

2023-01-13 | PXD039396 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data