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Series GSE125685 Query DataSets for GSE125685
Status Public on Jan 26, 2019
Title RNA-seq analysis of FACS-sorted control and Tsc2 cKO nociceptors with and without sciatic nerve crush
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary 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.
Overall design L4 dorsal root ganglia contralateral and ipsilateral to sciatic nerve crush were harvested 3 days post-injury from mice with Nav1.8-Cre mediated expression of GFP from Rosa26 locus as well as heterozgous (control) or homozygous (cKO) deletion of Tsc2 were FACS-sorted by GFP expression for 3 technical replicates of 100 cells each. RNA-seq data from 4 biological replicates was collected.
Contributor(s) Carlin D, Halevi A, Ewan EE, Moore A, Cavalli V
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Submission date Jan 25, 2019
Last update date Feb 11, 2019
Contact name Dan Carlin
Phone 314-362-0155
Organization name Washington University in St Louis
Department Neuroscience
Lab Valeria Cavalli
Street address 660 S Euclid Ave / Campus Box 8108
City St Louis
State/province MO
ZIP/Postal code 63110
Country USA
Platforms (1)
GPL21493 Illumina HiSeq 3000 (Mus musculus)
Samples (48)
GSM3580228 Uninjured control replicate 1A
GSM3580229 Uninjured control replicate 1B
GSM3580230 Uninjured control replicate 1C
BioProject PRJNA517164
SRA SRP182089

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Supplementary file Size Download File type/resource
GSE125685_RAW.tar 8.9 Mb (http)(custom) TAR (of TXT)
GSE125685_Tsc2cKO_SNI_DEseq2.csv.gz 598.5 Kb (ftp)(http) CSV
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Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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