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RNA-Seq of single cells from the mouse lumbar dorsal root ganglion

ABSTRACT: In order to establish a consensus catalog of dorsal rott ganglion cell types, we used comprehensive transcriptome analysis of single cells for unsupervised identification and molecular classification of sensory neurons independent of any a priori knowledge of sensory subtypes.

ORGANISM(S): Mus musculus

PROVIDER: GSE59739 | GEO | 2014/11/24

SECONDARY ACCESSION(S): PRJNA256268

REPOSITORIES: GEO

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RNA-Seq of single cells from the mouse lumbar dorsal root ganglion

Project description:In order to establish a consensus catalog of dorsal rott ganglion cell types, we used comprehensive transcriptome analysis of single cells for unsupervised identification and molecular classification of sensory neurons independent of any a priori knowledge of sensory subtypes. RNA-Seq was performed on 799 dissociated single cells dissected from the mouse lumbar dorsal root ganglion distributed over a total of nine 96-well plates

2014-11-24 | E-GEOD-59739 | biostudies-arrayexpress

Neuronal cell lines as model dorsal root ganglion neurons: a transcriptomic comparison

Project description:Aim: Dorsal root ganglion neuron-derived immortal cell lines including ND7/23 and F-11 cells have been used extensively as in vitro model systems of native peripheral sensory neurons. However, while it is clear that some sensory neuron-specific receptors and ion channels are present in these cell lines, a systematic comparison of the molecular targets expressed by these cell lines with intact peripheral neurons is lacking. Methods: we examined the expression of RNA transcripts in the human neuroblastoma-derived cell line, SH-SY5Y, and two dorsal root ganglion hybridoma cell lines, F-11 and ND7/23, using Illumina next-generation sequencing. Results: The expression profile of these three cell lines did not resemble any specific dorsal root ganglion neuron subclass. The cell lines lacked many markers for nociceptive sensory neurons, such as the transient receptor potential V1 gene, but expressed markers for both myelinated and unmyelinated neurons. Conclusion: This paper provides insights into the receptor repertoire expressed in common dorsal root ganglion neuron-derived cell lines, and illustrates the limits and potentials of these cell lines as tools for neuropharmacological exploration.

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Project description:The molecular identity of touch sensation is still largely unknown. We choose an extrem and very specific example, touch sensation of pennis glan, to study this problem. It's known from previous retro-grade labeling result that only one DRG in rat innervate the pennis glan, which makes it idea for microarry analysis. To clone the mechanosensory receptor specifically or highly expressed in the primary sensory neurons innervating pennis glan. There is a specific mechanosensory receptor in the primary sensory neurons innervating pennis glan. 1. Retrograde labeling the innervating nerve by DiI 2. Dissect DRGs, dissociate neurons and pick up the fluorescent cells. 3. Pool around 20 positive cells and 50 control cells (big diameter neuron and small to medium diameter neuron respectively), purify total RNA. 4. Two rounds of amplification 5. Affymetrix analysis Keywords: dorsal root ganglion, touch sensatiion

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