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

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

Project description: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.

2021-06-09 | GSE176350 | GEO

High-throughput mapping of single-neuron projection and molecular features by retrograde barcoded labelling [bulk RNA-seq]

Project description:Deciphering patterns of connectivity between neurons in the mammalian brain is a critical step toward understanding brain function. Conventional imaging based neuroanatomical tracing methods identify area-to-area or sparse neuron-to-neuron connectivity patterns, but with extremely limited throughput. Recently developed barcode-based connectomics methods can efficiently map large numbers of single-neuron projections, but linking these data to single-cell transcriptomics remains a challenge. Here, we established a retro-AAV barcode-based multiplexed tracing method called MERGE-seq (Multiplexed projection neuRons retroGrade barcodE sequencing), which is capable of simultaneously characterizing the projectome and transcriptome at the single neuron level. We uncovered dedicated and collateral projection patterns of ventromedial prefrontal cortex (vmPFC) neurons to five downstream targets (AI, DMS, BLA, MD and LH). We found that projection-defined vmPFC neurons are molecularly heterogeneous, which are composed of different neuronal subtypes. We further identified transcriptional signatures of various dedicated and bifurcated vmPFC neurons, and verified Pou3f1 as the marker gene of neurons sending collateral axons to DMS and LH. Finally, we fitted our single-neuron connectome/transcriptome data into a machine learning-based model and revealed groups of genes that were predictive of certain projection pattern. In summary, we have developed a new multiplexed technique whose paired connectome and gene expression data can help reveal organizational principles that form neural circuits and process information.

2024-02-25 | GSE210173 | GEO

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

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

| PRJNA682579 | ENA

High-throughput mapping of single-neuron projection and molecular features by retrograde barcoded labelling [scRNA-seq]

2024-02-25 | GSE210172 | GEO

Pathological consequences of chronic olfactory inflammation on neurite morphology of olfactory bulb projection neurons

Project description:The effects of chronic olfactory inflammation on the olfactory bulb (OB) were examined. Inflammation was induced in the olfactory epithelium by repeated unilateral intranasal administration of lipopolysaccharide (LPS). After 4 weeks, gene expression profiles were compared between ipislateral and contralateral OBs using RNA sequencing analysis.

2022-04-06 | GSE185945 | GEO

Pathological consequences of chronic olfactory inflammation on neurite morphology of olfactory bulb projection neurons

Project description:Pathological consequences of chronic olfactory inflammation on neurite morphology of olfactory bulb projection neurons

| PRJNA771410 | ENA