Culture-induced changes in blood-brain barrier transcriptome: implications for amino-acid transporters in vivo

Lyck, Ruth; Ruderisch, Nadine; Moll, Anton G; Steiner, Oliver; Cohen, Clemens D; Engelhardt, Britta; Makrides, Victoria; Verrey, Francois (2009). Culture-induced changes in blood-brain barrier transcriptome: implications for amino-acid transporters in vivo. Journal of cerebral blood flow and metabolism, 29(9), pp. 1491-502. New York, N.Y.: Nature Publishing Group 10.1038/jcbfm.2009.72

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Tight homeostatic control of brain amino acids (AA) depends on transport by solute carrier family 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 platelet endothelial cell adhesion molecule-1-positive (PECAM1(+)) 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 reverse transcription real-time PCR. Regardless of glial coculture, 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 physiologic functions, for example, tight barrier formation, and their consequent dedifferentiation in culture. Seven (4F2hc, Lat1, Taut, Snat3, Snat5, Xpct, and Cat1) of nine AA transporter mRNAs highly expressed in freshly isolated ppMBMECs were strongly downregulated for all cultures and two (Snat2 and Eaat3) were variably regulated. In contrast, five AA transporter mRNAs with low expression in ppMBMECs, including y(+)Lat2, xCT, and Snat1, were upregulated by culture. We hypothesized that the AA transporters highly expressed in ppMBMECs and downregulated in culture have a major in vivo function for BBB transendothelial transport.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Theodor Kocher Institute

UniBE Contributor:

Lyck, Ruth; Steiner, Oliver and Engelhardt, Britta

ISSN:

0271-678X

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 15:13

Last Modified:

04 May 2014 23:23

Publisher DOI:

10.1038/jcbfm.2009.72

PubMed ID:

19491922

Web of Science ID:

000269447600001

URI:

https://boris.unibe.ch/id/eprint/32142 (FactScience: 197069)

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