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.
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
Hélène
H
Legros
Séverine
S
Launay
Benoit Denis
BD
Roussel
Aurélie
A
Marcou-Labarre
Sébastien
S
Calbo
Julie
J
Catteau
Philippe
P
Leroux
Olivier
O
Boyer
Carine
C
Ali
Stéphane
S
Marret
Denis
D
Vivien
Vincent
V
Laudenbach
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