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Series GSE75668 Query DataSets for GSE75668
Status Public on Oct 24, 2016
Title RNA-seq of brain mural cells and microvascular fragments
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Purpose: Pericytes, the mural cells of blood microvessels, have come into focus as regulators of microvascular development and function, but due to paucity of defining markers, the identification and functional characterization of PC remain problematic, and reported data are often controversial. Here, we used a new approach for the isolation of mural cell from mouse brain in combination with RNA-sequencing (RNA-seq) and previously published vascular transcriptome data to assemble a state-of-the-art catalogue of brain mural cell-enriched gene transcripts.
Methods: We isolated double positive cells from the brain of Pdgfrb-eGFP/NG2-DsRed transgenic mice using FACS. Cells were lysed, RNA extracted and sequenced with next-generation sequencing (NGS). For comparison, we also determined the transcriptome of brain microvascular fragments (containing both endothelial cells and mural cells) isolated by mechanical tissue disintegration, collagenase digestion and immune-panning using anti-CD31 antibodies coupled to magnetic beads. The reads were aligned to the Ensembl mouse gene assembly (NCBIM37) using Tophat2 software (version 2.0.4). The duplicated reads were removed using the picard tool (version 1.92). To identify the genes significantly enriched in the pericyte samples as compared with microvascular samples, statistical tests were performed using the Cufflinks tool (version 2.2.1)
Results: The result showed that mRNA transcripts representing 856 different genes were enriched more than two-fold in FACS isolated Pdgfrb-eGFP/NG2-DsRed double positive cells compared with whole microvascular fragments (False Discovery Rate < 0.05)
 
Overall design The RNA from three FACS sorted brain mural cell samples and three whole brain microvascular samples isolated from three animals were processed and sequenced on the Illumina HiSeq 2500 platform in the sequencing facility in Uppsala University.
 
Contributor(s) Betsholtz C
Citation(s) 27725773
Submission date Dec 03, 2015
Last update date May 15, 2019
Contact name L. H.
Organization name Uppsala University
Street address Dag Hammarskjölds Väg 20
City Uppsala
ZIP/Postal code 751 85
Country Sweden
 
Platforms (1)
GPL17021 Illumina HiSeq 2500 (Mus musculus)
Samples (6)
GSM1963121 Brain microvascular fragments 1
GSM1963122 Brain microvascular fragments 2
GSM1963123 Brain microvascular fragments 3
Relations
BioProject PRJNA305031
SRA SRP066974

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Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE75668_RAW.tar 1.0 Mb (http)(custom) TAR (of TXT)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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