title: Purification of Mouse Brain Vessels.
abstract: In the brain, most of the vascular system consists of a selective barrier, the blood-brain barrier (BBB) that regulates the exchange of molecules and immune cells between the brain and the blood. Moreover, the huge neuronal metabolic demand requires a moment-to-moment regulation of blood flow. Notably, abnormalities of these regulations are etiological hallmarks of most brain pathologies; including glioblastoma, stroke, edema, epilepsy, degenerative diseases (ex: Parkinson's disease, Alzheimer's disease), brain tumors, as well as inflammatory conditions such as multiple sclerosis, meningitis and sepsis-induced brain dysfunctions. Thus, understanding the signaling events modulating the cerebrovascular physiology is a major challenge. Much insight into the cellular and molecular properties of the various cell types that compose the cerebrovascular system can be gained from primary culture or cell sorting from freshly dissociated brain tissue. However, properties such as cell polarity, morphology and intercellular relationships are not maintained in such preparations. The protocol that we describe here is designed to purify brain vessel fragments, whilst maintaining structural integrity. We show that isolated vessels consist of endothelial cells sealed by tight junctions that are surrounded by a continuous basal lamina. Pericytes, smooth muscle cells as well as the perivascular astrocyte endfeet membranes remain attached to the endothelial layer. Finally, we describe how to perform immunostaining experiments on purified brain vessels.
authors.affiliation: Collège de France, Center for Interdisciplinary Research in Biology (CIRB), Centre National de la Recherche Scientifique CNRS, Unité Mixte de Recherche 7241, Institut National de la Santé et de la Recherche Médicale; Centre Interdisciplinaire de Recherche en Biologie; MEMOLIFE Laboratory of Excellence and Paris Science, Lettre Research University.
authors.forename: Anne-Cécile
authors.initials: AC
authors.lastname: Boulay
authors.affiliation: Université Paris Descartes, Faculté de Pharmacie, Université Paris Diderot.
authors.forename: Bruno
authors.initials: B
authors.lastname: Saubaméa
authors.affiliation: Université Paris Descartes, Faculté de Pharmacie, Université Paris Diderot.
authors.forename: Xavier
authors.initials: X
authors.lastname: Declèves
authors.affiliation: Collège de France, Center for Interdisciplinary Research in Biology (CIRB), Centre National de la Recherche Scientifique CNRS, Unité Mixte de Recherche 7241, Institut National de la Santé et de la Recherche Médicale; Centre Interdisciplinaire de Recherche en Biologie; MEMOLIFE Laboratory of Excellence and Paris Science, Lettre Research University; martine.cohen-salmon@college-de-france.fr.
authors.forename: Martine
authors.initials: M
authors.lastname: Cohen-Salmon
mesh_terms: D000818:Animals; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000166:cytology*; D001921:Brain / Q000098:blood supply*; D003584:Cytological Techniques / Q000379:methods; D042783:Endothelial Cells / Q000166:cytology; D051379:Mice; D020286:Pericytes / Q000166:cytology; D015398:Signal Transduction; D019108:Tight Junctions / Q000502:physiology