Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Sorted cells_PS2APP brains_7/13mo

ABSTRACT: Mice of indicated ages and genotypes were perfused and their brains dissected and dissociated. Cells were fixed, immunolabeled and FACS sorted. RNA was extracted from neuron, astrocyte, and microglial cell populations. Typical RIN=4-5 for neurons, 6-8 for astrocytes, and 5-7 for microglia. Typical RNA yields ~100ng for neurons, ~20ng for microglia, and ~10ng for astrocytes. cDNA was generated from up to 25 ng of total RNA using Nugen'™s RNA-Seq method for low-input RNA samples, Ovation RNA-Seq System V2 (NuGEN). (Per manufacturers instructions, total RNA was neither depleted of rRNA nor polyA-selected.) 1 ug of sheared cDNA was taken into further processing, starting at end repair step, using Illumina'™s TruSeq RNA Sample Preparation Kit v2 (Illumina). The 'SAMPLE_ID'; sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0006149 Astrocytes, microglia and neurons were sorted from 7- or 13-month old PS2APP or non-transgenic mice, 4 <= n <= 7 per group.

ORGANISM(S): Mus musculus

SUBMITTER: Brad Friedman 

PROVIDER: E-GEOD-75431 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A common approach to understanding neurodegenerative disease is comparing gene expression in diseased versus healthy tissues. We illustrate that expression profiles derived from whole tissue RNA highly reflect the degenerating tissues' altered cellular composition, not necessarily transcriptional regulation. To accurately understand transcriptional changes that accompany neuropathology, we acutely purify neurons, astrocytes and microglia from single adult mouse brains and analyse their transcrip  ...[more]

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