Ontology highlight
ABSTRACT:
PROVIDER: PRJNA595403 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR10680893_1.fastq.gz | Fastqsanger.gz | |||
| SRR10680893_2.fastq.gz | Fastqsanger.gz | |||
| SRR10680894_1.fastq.gz | Fastqsanger.gz | |||
| SRR10680894_2.fastq.gz | Fastqsanger.gz | |||
| SRR10680895_1.fastq.gz | Fastqsanger.gz |
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Project description:The richness of our somatosensory experience is reflected in the functional diversity of somatic sensory neurons. Single-cell sequencing (scRNA) of sensory neurons has revealed a molecular basis for such diversity1–3. However, sensory neuron diversity has yet to be captured at the level of the proteome. Here, we combined electrophysiology with deep visual proteomics (DVP)4 to quantify over 6,000 proteins from phenotypically-defined sensory neurons in mice and identified proteomic markers of sensory neuron subtypes. Comparative analysis revealed both concordance and meaningful divergence between transcriptomes and proteomes. We further show that up to 3,000 proteins can be quantified from one-fourth of a single neuron, demonstrating subset-specific protein signatures. In culture, nociceptive neurons can be acutely sensitized to mechanical stimuli by nerve growth factor (NGF) which normally drives inflam-matory pain in vivo5. Indeed, overnight exposure of peptidergic nociceptors to NGF and a pro-tein kinase C (PKC) activator produced functional sensitization associated with proteome changes. Functional knockdown experiments identified the up-regulated B3GNT2 enzyme as a potential effector of nociceptor sensitization. In summary, we present a high-resolution proteomic resource linking molecular identity to function, enabling the discovery of mechanisms un-derlying somatic sensation and pain sensitization.
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Project description:Amyotrophic lateral sclerosis (ALS) is primarily characterized by motor neuron degeneration, but accumulating evidence suggests that sensory neurons are also affected. To investigate early sensory neuron dysfunction in ALS, we performed bulk RNA sequencing of dorsal root ganglia (DRG) isolated from SOD1-G93A transgenic mice and wild-type controls at disease onset. This dataset provides transcriptomic profiles that reveal molecular alterations associated with ALS-related sensory neuron pathology.
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Project description:Sensory neurons are distinguished by distinct signaling networks and receptive characteristics. Thus, sensory neuron types can be defined by linking transcriptome-based neuron typing with the sensory phenotypes. Here we classify somatosensory neurons of the mouse dorsal root ganglion (DRG) by high-coverage single-cell RNA-sequencing (10 950 ± 1 218 genes per neuron) and neuron size-based hierarchical clustering. Moreover, single DRG neurons responding to cutaneous stimuli are recorded using an in vivo whole-cell patch clamp technique and classified by neuron-type genetic markers. Small diameter DRG neurons are classified into one type of low-threshold mechanoreceptor and five types of mechanoheat nociceptors (MHNs). Each of the MHN types is further categorized into two subtypes. Large DRG neurons are categorized into four types, including neurexophilin 1-expressing MHNs and mechanical nociceptors (MNs) expressing BAI1-associated protein 2-like 1 (Baiap2l1). Mechanoreceptors expressing trafficking protein particle complex 3-like and Baiap2l1-marked MNs are subdivided into two subtypes each. These results provide a new system for cataloging somatosensory neurons and their transcriptome databases. RNA-seq of mRNA levels in 197 individual DRG neurons We performed RNA-seq on total 203 individual DRG neurons. Six of them were not qualified and thus, were excluded for further analysis. To evaluate the quality of RNA-seq, we randomly devided No.72 neurons into two parts and performed RNA-seq seperately. Thus, we had 204 individual samples from 203 individual DRG and 198 individual qualified samples from 197 individual DRG. To evaluate the homogeneity of RNA-seq data from different mice at the same age just as used, we performed RNA-seq on 5 single DRG from different mice. Here, these data from DRG were also considered as experimental control. The 'DRG_neurons_RNA_Seq.txt' contains processed data for 204 samples and 'DRG_RNA_Seq.txt' for 5 samples.
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Project description:Cultured adult mouse dorsal root ganglia (DRG) cells exhibit glial sensory progenitor properties in vitro. Therefore, they might be a good starter cell for reprogramming into sensory neurons. Here, we infected them with the retroviral vector Neurog2-Neurog1-DsRed to induce sensory neuron development and analyzed by scRNAseq at 14 days post infection whether infected cells show properties of sensory neurons such as nociceptors. After 10x Genomics, data analysis of 4,549 individual cells indicated the generation of neurons, but at an immature cell state.
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Project description:Chronic tendinopathy is typified by persistent tendon-associated pain, transmitted by local nociceptive neurons. However, the function of somatosensory neurons in the development of tendinopathy is entirely unknown. Here, we show that sensory neurons grow into the tendon proper across models of chronic tendinopathy. Three complementary surgical and transgenic mice models of disrupted sensory nerve growth were next utilized. Conditional deletion of Nerve growth factor (NGF) in macrophages (Ngf Csfr1) or inactivation of its high affinity receptor Tropomyosin receptor kinase A (TrkA) on sensory neurons led to severely worsened tendinopathy. A sensory-only sural nerve denervation model phenocopied these results, including heightened macrophage infiltration and tenocyte apoptosis. Single-cell RNA sequencing (scRNA-seq) of tendinous tissue identified defective tenocyte differentiation and altered macrophage migration and polarization with surgical denervation. Retrograde neuronal tracing in combination with scRNA-seq of corresponding dorsal root ganglia (DRG) tissues identified the profile of tendon-specific innervation, which included CGRP+ nociceptors among other neuron types. Finally, neuron-tendon interaction analyses implicated neuron-derived fibroblast growth factor 1 (FGF1) as a potent regulator of tendon repair, a finding experimental confirmed with tendon organ culture. Collectively, our findings demonstrate that peripheral afferent neural networks induce a protective effect in chronic tendinopathy by secreting FGF1, and that targeting this pathway may offer therapeutic strategies to enhance tendon repair.
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Project description:Peripheral Sensory Neuron EC-tagging experiments
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Project description:Identify neuron-specific ubiquitin targets of Infected Cell Protein 0 (ICP0) in adult sensory neurons.
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Project description:Sensory neurons are distinguished by distinct signaling networks and receptive characteristics. Thus, sensory neuron types can be defined by linking transcriptome-based neuron typing with the sensory phenotypes. Here we classify somatosensory neurons of the mouse dorsal root ganglion (DRG) by high-coverage single-cell RNA-sequencing (10 950 ± 1 218 genes per neuron) and neuron size-based hierarchical clustering. Moreover, single DRG neurons responding to cutaneous stimuli are recorded using an in vivo whole-cell patch clamp technique and classified by neuron-type genetic markers. Small diameter DRG neurons are classified into one type of low-threshold mechanoreceptor and five types of mechanoheat nociceptors (MHNs). Each of the MHN types is further categorized into two subtypes. Large DRG neurons are categorized into four types, including neurexophilin 1-expressing MHNs and mechanical nociceptors (MNs) expressing BAI1-associated protein 2-like 1 (Baiap2l1). Mechanoreceptors expressing trafficking protein particle complex 3-like and Baiap2l1-marked MNs are subdivided into two subtypes each. These results provide a new system for cataloging somatosensory neurons and their transcriptome databases.
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