Newborn- and adult-derived brain microvascular endothelial cells show age-related differences in phenotype and glutamate-evoked protease release

• Published in: Journal of cerebral blood flow and metabolism 2009-06, Vol.29 (6), p.1146-1158
• Main Authors: Legros, Hélène, Launay, Séverine, Roussel, Benoit Denis, Marcou-Labarre, Aurélie, Calbo, Sébastien, Catteau, Julie, Leroux, Philippe, Boyer, Olivier, Ali, Carine, Marret, Stéphane, Vivien, Denis, Laudenbach, Vincent
• Format: Article
• Language: English
• Subjects: Aging - physiology; Animals; Animals, Newborn; Biological and medical sciences; Biomarkers; Brain - blood supply; Brain - cytology; Brain - drug effects; Brain - enzymology; Cell Shape; Cells, Cultured; Cerebral circulation. Blood-brain barrier. Choroid plexus. Cerebrospinal fluid. Circumventricular organ. Meninges; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Cells - enzymology; Endothelial Cells - secretion; Excitatory Amino Acid Transporter 2 - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation - drug effects; Gene Expression Regulation - genetics; Glutamic Acid - pharmacology; Matrix Metalloproteinase 2 - metabolism; Matrix Metalloproteinase 9 - metabolism; Medical sciences; Mice; Microvessels - cytology; Microvessels - drug effects; Microvessels - enzymology; Microvessels - secretion; Monocarboxylic Acid Transporters - metabolism; Neurology; Phenotype; Protein Subunits - genetics; Protein Subunits - metabolism; Receptors, N-Methyl-D-Aspartate - metabolism; Symporters - metabolism; Tissue Culture Techniques; Vascular diseases and vascular malformations of the nervous system; Vertebrates: nervous system and sense organs
• ISSN: 0271-678X; 1559-7016
• DOI: 10.1038/jcbfm.2009.39

Few data are available on the involvement of brain microvascular endothelial cells (BMECs) in excitotoxic neonatal brain lesions. Therefore, we developed an original approach for investigating mouse-derived BMECs in vitro. We hypothesized that newborn and adult BMEC cultures would show age-related differences in phenotype and sensitivity to glutamate. Expression of the monocarboxylate transporter, MCT1, was higher in neonatal than in adult BMECs, whereas expression of the glucose transporter, GLUT1, was higher in adult than in neonatal BMECs that overexpressed the N-methyl-D-aspartate receptor NR1 subunit (NMDAR1) compared with adult BMECs. The ability of neonatal and adult BMECs to be activated by glutamate was confirmed through intracellular calcium ([Ca2+]i) recording. The glutamate-induced [Ca2+]i increase was blocked by the selective NMDAR antagonist, MK-801. Significant glutamate-evoked concentration-dependent release of tissue-type plasminogen activator (t-PA) and matrix metalloproteinases (MMPs) activities was found in supernatants of neonatal, but not in adult BMECs. The glutamate-mediated release of t-PA, MMP-2, and MMP-9 proteolytic activities in neonatal BMECs was blocked by MK-801. Conceivably, this protease release from neonatal BMECs may participate in neonatal brain lesions.