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Series GSE14375 Query DataSets for GSE14375
Status Public on Jun 01, 2009
Title Culture induced changes in Blood-Brain Barrier transcriptome: implications for amino acid transporters in vivo
Organism Mus musculus
Experiment type Expression profiling by array
Summary Tight homeostatic control of brain amino acids (AA) depends on transport via solute family carrier proteins expressed by the Blood-Brain Barrier (BBB) microvascular endothelial cells (BMEC). To characterize the mouse BMEC transcriptome and probe culture-induced changes microarray analyses of PECAM-1+ endothelial cells (ppMBMECs) were compared with primary MBMECs (pMBMEC) cultured in the presence or absence of glial cells, and with b.End5 endothelioma cell-line. Selected cell marker and AA transporter mRNA levels were further verified by real-time RT PCR.
Regardless of glial co-culture expression of a large subset of genes was strongly altered by a brief culture step. This is consistent with the known dependence of BMECs on in vivo interactions to maintain physiological functions, e.g. tight barrier formation, and their consequent de-differentiation in culture. Seven (4F2hc, Lat1, Taut, Snat3, Snat5, Xpct, Cat1) of nine highly in vivo expressed AA transporter mRNAs were strongly down-regulated for all cultures and two (Snat2, Eaat3) were variably regulated. In contrast, five AA transporter mRNAs with low in vivo expression, including y+Lat2, xCT, and Snat1, were strongly up-regulated by culture. We hypothesized that the AA transporters highly expressed in ppMBMECs and strongly down-regulated in culture play a major in vivo role for BBB transendothelial transport.

Keywords: culture vs non-cultured
 
Overall design RNA was prepared from 4 sources of endothelial cells: 1) isolated mouse brain microvascular endothelial cells that were cultured in transwells with and 2) without non-contact co-culture with glial cultures or 3) further purified using anti-PECAM1 magnetic bead sorting and 4) the endothelioma b.End5 cell-line.
 
Contributor(s) Verrey F, Makrides V, Lyck R, Ruderisch N, Cohen CD, Moll AG, Engelhardt B
Citation(s) 19491922
Submission date Jan 12, 2009
Last update date Mar 20, 2012
Contact name Francois Verrey
E-mail(s) verrey@access.uzh.ch
Phone +41 44 635 50 44
Fax +41 44 635 68 14
Organization name University of Zurich
Department Institute of Physiology
Street address Winterthurerstr. 190
City Zurich
ZIP/Postal code CH-8057
Country Switzerland
 
Platforms (1)
GPL8059 Mouse Genome 430 2.0 Array [Mm430_Mm_REFSEQ (v10) CDF]
Samples (12)
GSM359429 non-cultured ppMBMEC - 1
GSM359430 non-cultured ppMBMEC - 2
GSM359431 non-cultured ppMBMEC - 3
Relations
BioProject PRJNA111419

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE14375_RAW.tar 54.8 Mb (http)(custom) TAR (of CEL, CHP)
Raw data provided as supplementary file
Processed data included within Sample table
Processed data provided as supplementary file

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