Newborn- and adult-derived brain microvascular endothelial cells show age-related differences in phenotype and glutamate-evoked protease release.
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.
Legros
Hélène
H
EA 4309 Neovasc Microvascular Endothelium and Neonatal Brain Lesions, IHURBM, IFRMP 23, School of Medicine and Pharmacy, University of Rouen, Rouen, France. helene.legros@univ-rouen.fr
Launay
Séverine
S
Roussel
Benoit Denis
BD
Marcou-Labarre
Aurélie
A
Calbo
Sébastien
S
Catteau
Julie
J
Leroux
Philippe
P
Boyer
Olivier
O
Ali
Carine
C
Marret
Stéphane
S
Vivien
Denis
D
Laudenbach
Vincent
V
D000375:Aging / Q000502:physiology*; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers; D001921:Brain / Q000098:blood supply* / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D048430:Cell Shape; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology* / Q000378:metabolism; D027342:Excitatory Amino Acid Transporter 2 / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D018698:Glutamic Acid / Q000494:pharmacology*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D055806:Microvessels / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D027501:Monocarboxylic Acid Transporters / Q000378:metabolism; D010641:Phenotype; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D027981:Symporters / Q000378:metabolism; D046509:Tissue Culture Techniques; D000099233:Monocarboxylate Transport Protein 1
D015415:Biomarkers; D027342:Excitatory Amino Acid Transporter 2; D018698:Glutamic Acid; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D027501:Monocarboxylic Acid Transporters; D021122:Protein Subunits; D016194:Receptors, N-Methyl-D-Aspartate; D027981:Symporters; D000099233:Monocarboxylate Transport Protein 1