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FACS purified Nav1.8 lineage sensory neuron whole transcriptome changes following peripheral inflammation (CFA) or sciatic nerve injury (SNI).

ABSTRACT: FACS purified Nav1.8 lineage sensory neuron whole transcriptome changes following peripheral inflammation (CFA) or sciatic nerve injury (SNI).

PROVIDER: PRJNA916269 | ENA |

REPOSITORIES: ENA

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FACS purified Nav1.8 lineage sensory neuron whole transcriptome changes following peripheral inflammation (CFA) or sciatic nerve injury (SNI).

Project description:The goal of this study was to analyze global gene expression in FACS purified Nav1.8 lineage sensory neurons, which include nociceptor neurons that detect damaging/noxious stimuli, following peripheral inflammation by intraplantar injection of Complete Freund's Adjuvant (CFA) or Sciatic Nerve Injury (SNI) by nerve transection. Nav1.8 Trangsgenic TdTomato+ neurons were purified from Lumbar L4-L6 dorsal root ganglia (DRG) by flow cytometry from mice on the ipsilateral or contralateral sides, following Complete Freund's Adjuvant injection (day 1) or sciatic nerve transection (day 5). Neurons were then analyzed for transcriptional gene expression by microarray analysis.

2023-01-30 | GSE221834 | GEO

Adult and Neonatal dorsal horn gene expression 7 day post sciatic nerve SNI injury

Project description:As rats do not develop neuropathic pain like hypersensitivity as neonates post nerve injury but do as adults we have used these arrays to help define the processes involved in this process. Rat spinal cord (ipsilateral dorsal horn) was assayed 7 days post SNI injury to the sciatic nerve relative to sham injury. Two age groups of animals were tested Neonates (P10) and Adult (8-12wks). Keywords: Two way analysis of differential regulation

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RNA sequencing of whole sciatic Dorsal Root Ganglia (DRG) from young and aged mice

Project description:Purpose: compare the gene expression in whole sciatic DRG of aged mice preceding and after sciatic nerve injury (SNI) compared to the young Results: we found ageing-dependent gene expression changes repressing axonal regeneration following sciatic nerve injury

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RNA-seq analysis of FACS-sorted control and Tsc2 cKO nociceptors with and without sciatic nerve crush

Project description:The goals of this study are to compare the transcriptomes of control heterozygous and Tuberous sclerosis 2 (Tsc2) homozygous nociceptors 3 days after sciatic nerve injury that have been enriched by fluorescence-associated cell sorting (FACS) with the aim of identifying differences in genes associated with axon regeneration. Nav1.8-Cre transgenic mice were used to delete Tsc2 as well as express Green fluorescent protein from the Rosa26 locus. Tsc2 deletion constitutively activates mTORC1 signaling cell autonomously. Dissociated dorsal root ganglia from adult mice were FACS-sorted for GFP and analyzed by RNA-seq. We find that the expression of a number of regeneration-associated transcription factors including cJun, Atf3, Sox11 and other are upregulated in Tsc2 mutant neurons in the absence of an injury. Additionally, the target genes of several other normally expressed regeneration-associated transcription factors are differentially expressed in Tsc2 mutant neurons, such as Creb1 and Klf’s. We conclude Tsc2 deletion and consequent mTORC1 activation establishes a pro-regenerative gene expression profile in nociceptors in the absence of an injury.

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Expression data from FACS purified nociceptor neurons from peripheral sensory ganglia

Project description:The goal of this study was to analyze global gene expression in specific populations of nociceptor sensory neurons, the neurons that detect damaging/noxious stimuli. The dorsal root ganglia (DRG), trigeminal ganglia, and nodose ganglia are anatomically distinct peripheral sensory ganglia that contain nociceptors which innervate skin, gut, lungs, and other distinct organ tissues. We used flow cytometry to purify nociceptors from these ganglia and profiled their global gene expression signatures to compare gene expression between these different anatomically distinct nociceptors. Nav1.8-Cre were bred with Rosa26-TdTomato to generate Nav1.8-Cre/R26-TdTomato reporter progeny, where all peripheral nociceptor neurons are genetically marked with red fluroescence due to specific expression of the TTX- resistant sodium channel Nav1.8. Lumbar region dorsal root ganglia (DRG), trigeminal ganglia, and nodose ganglia were dissected from mice (3 mice were pooled/sample). Highly red fluorescent neurons were Facs purified, RNA extracted, and processed for microarray analysis.

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Ascending dorsal column sensory neurons respond to spinal cord injury and downregulate genes related to lipid metabolism

Project description:The goal of this study was to compare the transcriptional effects of sciatic nerve injury and spinal cord injury on lumbar dorsal root ganglion (DRG) and FACS-sorted dorsal column (DC) sensory neurons. We performed RNA-seq of whole DRG from naïve and spinal cord-injured (SCI) mice (1dpi) and compared this with previously published data for sciatic nerve transection. In order to assess changes specifically in DC neurons, we performed RNA-seq from FACS-sorted DC neurons from Thy1-YFP16 transgenic mice in naïve, sciatic nerve injured (SNI), and SCI (1 and 3dpi). We found that DC neurons alter their transcriptome after SCI, but that gene changes after SCI mostly differ from SNI. These transcriptional differences may reflect both growth promoting and growth inhibitory effects on axon regeneration after SCI.

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Project description:As rats do not develop neuropathic pain like hypersensitivity as neonates post nerve injury but do as adults we have used these arrays to help define the processes involved in this process. Rat spinal cord (ipsilateral dorsal horn) was assayed 7 days post SNI injury to the sciatic nerve relative to sham injury. Two age groups of animals were tested Neonates (P10) and Adult (8-12wks). Experiment Overall Design: Six biologically indepenedent arrays were hybridized per assay point. Dorsal horn total RNA was prepared using standard Affymetrix protocols. Affymetrix Rat Expression 230A array used.

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