Dataset Information


Culture induced changes in Blood-Brain Barrier transcriptome: implications for amino acid transporters in vivo

ABSTRACT: 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.

INSTRUMENT(S): Mouse Genome 430 2.0 Array [Mm430_Mm_REFSEQ (v10) CDF]

ORGANISM(S): Mus musculus  

SUBMITTER: Francois Verrey  

PROVIDER: GSE14375 | GEO | 2009-06-01



Similar Datasets

2010-05-18 | E-GEOD-14375 | ArrayExpress
| GSE94064 | GEO
| GSE97575 | GEO
| GSE85657 | GEO
2016-03-17 | E-GEOD-76530 | ArrayExpress
2016-03-17 | E-GEOD-76528 | ArrayExpress
| GSE74052 | GEO
2010-10-21 | E-GEOD-15892 | ArrayExpress
2010-10-21 | GSE15892 | GEO
2012-09-29 | E-GEOD-41211 | ArrayExpress