PXD002759

PXD002759 is an original dataset announced via ProteomeXchange.

Title	Development and evaluation of a SILAC method for proteomic analysis of rat primary microglia
Description	Microglia play important and dynamic roles in mediating a variety of physiological and pathological processes during the development, normal function, and degeneration of the central nervous system. We report here the development of a SILAC-labeled model system suitable for mass spectrometry-based proteomic profiling in primary rat microglia. A mixed primary glia culture was first established by isolating newborn brain cells from Sprague-Dawley rats and maintaining the culture in a fetal bovine serum-supplemented and chemically defined media containing either the regular amino acids (light label media) or stable isotope-labeled amino acids (heavy label media). After isolation of microglia from the mixed glia cultures, mass spectrometric analysis indicated that the incorporation efficiency of the heavy-labeled amino acids was between 70-80%. Upon stimulation with bacterial endotoxin lipopolysaccharide to induce classical (M1) activation, the light- and heavy-labeled primary microglia responded indistinguishably as judged by the up-regulation of inducible nitric oxide synthase. More important, the relative quantitation accuracy and proteome coverage obtained by the reported SILAC model allowed for identification of classical microglial activation markers and pathways involved in the proinflammatory response of microglia as determined by bioinformatic analysis. The establishment of this viable primary microglia SILAC model will further expand our capacity for global scale proteomic profiling of pathways mediating the multifaceted microglial responses in various physiological and pathological processes.
HostingRepository	PRIDE
AnnounceDate	2017-01-17
AnnouncementXML	Submission_2017-01-17_07:29:20.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD002759
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Ashley Cochran
SpeciesList	scientific name: Rattus norvegicus (Rat); NCBI TaxID: 10116;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2015-08-18 05:35:52	ID requested
1	2016-04-07 01:52:50	announced
⏵ 2	2017-01-17 07:29:23	announced	Updated project metadata.

Zhang P, Culver-Cochran AE, Stevens SM, Liu B, Characterization of a SILAC method for proteomic analysis of primary rat microglia. Proteomics, 16(9):1341-6(2016) [pubmed]

curator keyword: Biological

submitter keyword: primary, microglia, SILAC, rat

Stanley M. Stevens, Jr.
contact affiliation	Department of Cell Biology, Microbiology, and Molecular Biology University of South Florida Tampa, FL USA
contact email	smstevens@usf.edu
lab head
Ashley Cochran
contact affiliation	University of South Florida
contact email	aecochran@usf.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/04/PXD002759

PRIDE project URI

• PRIDE
  1. PXD002759
     1. Label: PRIDE project
     2. Name: Development and evaluation of a SILAC method for proteomic analysis of rat primary microglia