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ScRNA-seq of electrophysiologically characterized DRG neurons reveals subtype-specific and maturational differences in gene expression

ABSTRACT: scRNA-seq of electrophysiologically characterized DRG neurons reveals subtype-specific and maturational differences in gene expression

PROVIDER: PRJNA645297 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12192100.fastq.gz	Fastqsanger.gz
	SRR12192101.fastq.gz	Fastqsanger.gz
	SRR12192102.fastq.gz	Fastqsanger.gz
	SRR12192072.fastq.gz	Fastqsanger.gz
	SRR12192073.fastq.gz	Fastqsanger.gz

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scRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals maturational differences in gene expression

Project description:scRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals maturational differences in gene expression

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scRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals maturational differences in gene expression

Project description:Cultured dorsal root ganglia neurons of 4 and 12 weeks old C57Bl/6 mice were first characterized as mechanosensitive (showed rapidly adapting mechanically actived currents in whole-cell votage clamp) and as mechanoreceptors (evoked action potential waveform in whole-cell current clamp). Next, cells were subjected to RNA-seq and sequences were analyzed bioinformatically. We found maturatinal changes comprising age-dependent differential gene expression in these somatosensory mechanoreceptors. The found differences call for tight age-matching of young mice in research and underline found electrophysiological and behavioural maturational changes in somatosensation, see Michel, Niklas and Narayanan, Pratibha and Shomroni, Orr and Schmidt, Manuela, Cutaneous Touch and Piezo2-Mediated Mechanotransduction Decline Throughout Mouse Maturation Until Adulthood. CELL-REPORTS-D-20-00425. Available at SSRN: https://ssrn.com/abstract=3536401 or http://dx.doi.org/10.2139/ssrn.3536401

2020-07-21 | GSE154159 | GEO

scRNA-seq of electrophysiologically characterized DRG neurons reveals subtype-specific and maturational differences in gene expression

Project description:Cultured dorsal root ganglia neurons of 4 and 12 weeks old C57Bl/6 mice were first characterized as mechanosensitive (showed rapidly adapting mechanically actived currents in whole-cell votage clamp or not (non-responders)) and as either mechanoreceptors or nociceptors (evoked action potential waveform in whole-cell current clamp). Next, cells were subjected to RNA-seq and sequences were analyzed bioinformatically. We found maturatinal changes comprising subtype-specific and age-dependent differential gene expression in these somatosensory neurons. See Michel, Niklas and Narayanan, Pratibha and Shomroni, Orr and Schmidt, Manuela, Cutaneous Touch and Piezo2-Mediated Mechanotransduction Decline Throughout Mouse Maturation Until Adulthood. CELL-REPORTS-D-20-00425. Available at SSRN: https://ssrn.com/abstract=3536401 or http://dx.doi.org/10.2139/ssrn.3536401

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scRNA-sequencing reveals subtype-specific transcriptomic perturbations in DRG neurons of PirtEGFPf mice in neuropathic pain condition

Project description:Functionally distinct subtypes/clusters of dorsal root ganglion (DRG) neurons may play different roles in nerve regeneration and pain. However, details about their transcriptomic changes under neuropathic pain conditions remain unclear. Chronic constriction injury (CCI) of the sciatic nerve represents a well-established model of neuropathic pain, and we conducted single-cell RNA-sequencing (scRNA-seq) to characterize subtype-specific perturbations of transcriptomes in lumbar DRG neurons on day 7 post-CCI. By using PirtEGFPf mice that selectively express an enhanced green fluorescent protein in DRG neurons, we established a highly efficient purification process to enrich neurons for scRNA-seq. We observed the emergence of four prominent CCI-induced clusters and a loss of marker genes in injured neurons. Importantly, a portion of injured neurons from several clusters were spared from injury-induced identity loss, suggesting subtype-specific transcriptomic changes in injured neurons. Moreover, uninjured neurons, which are necessary for mediating the evoked pain, also demonstrated cell-type-specific transcriptomic perturbations in these clusters, but not in others. Notably, male and female mice showed differential transcriptomic changes in multiple neuronal clusters after CCI, suggesting transcriptomic sexual dimorphism in DRG neurons after nerve injury. Using Fgf3 as a proof-of-principle, RNAscope study provided further evidence of increased Fgf3 in injured neurons after CCI, supporting scRNA-seq analysis, and calcium imaging study unraveled a functional role of Fgf3 in neuronal excitability. These findings may contribute to the identification of new target genes and the development of DRG neuron cell-type-specific therapies for optimizing neuropathic pain treatment and nerve regeneration.

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Project description:In this study, we constructed a tissue-engineered DRG-CSC (corneal stromal cells) co-cultured model (DS model), a CSC monoculture model (S model), and a DRG monoculture model (D model). Chicken DRGs were extracted from E7-E10 chicken embryo. Human CSCs were extracted in stromal lenticules from small incision lenticule extraction (SMILE). Transcriptional profiling of DRG in the DS model (DS-D), DRG in the D model(D-D), CSCs in the DS model (DS-S), CSCs in the S model (S-S) was analyzed. After 7 days of culture, RNA of each group was extracted.

2021-06-29 | GSE179020 | GEO

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