title: Basic fibroblast growth factor modulates density of blood vessels and preserves tight junctions in organotypic cortical cultures of mice: a new in vitro model of the blood-brain barrier.
abstract: This study was performed to examine the maintenance of blood vessels in vitro in cortical organotypic slice cultures of mice with special emphasis on basic fibroblast growth factor (FGF-2), which is known to promote angiogenesis and to preserve the integrity of the blood-brain barrier. Slices of neonatal day 3 or 4 mouse brain were maintained for 3, 7, or 10 d in vitro (DIV) under standard culture conditions or in the presence of FGF-2. Immunohistochemistry for factor VIII-related antigen or laminin revealed a relative low number of blood vessels under standard conditions. In contrast, moderate FGF-2 concentrations increased the number of vessels: with 0.5 ng/ml FGF-2 it was 1.4-fold higher after DIV 3 or 1.5-fold after DIV 7 compared with controls; with 5 ng/ml it was almost doubled in both cases. With an excess of 50 ng/ml, FGF-2 vessels were reduced after DIV 3 or similar to controls after DIV 7. FGF receptor 1 was preferentially found on endothelial cells; its immunolabeling was reduced in the presence of the ligand. Cell death detected by an ethidium bromide analog or the apoptosis marker caspase-3 was barely detectable during the 10 d culture period. Immunolabeling of the tight junction proteins ZO-1 (zonula occludens protein 1), occludin, claudin-5, and claudin-3 revealed evidence for structural integrity of the blood-brain barrier in the presence of moderate FGF-2 concentrations. In conclusion, FGF-2 maintains blood vessels in vitro and preserves the composition of the tight junction. Hence, we propose FGF-2-treated organotypic cortical slices as a new tool for mechanistic studies of the blood-brain barrier.
authors: Section of Neuroanatomy, Institute of Anatomy, University of Basel, CH-4056 Basel, Switzerland. kerstin.bendfeldt@unibas.ch
authors: Kerstin
authors: K
authors: Bendfeldt
authors: Vesna
authors: V
authors: Radojevic
authors: Josef
authors: J
authors: Kapfhammer
authors: Cordula
authors: C
authors: Nitsch
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001808:Blood Vessels / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D001812:Blood-Brain Barrier / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000098:blood supply* / Q000166:cytology / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology* / Q000502:physiology; D051379:Mice; D009924:Organ Culture Techniques; D019108:Tight Junctions / Q000187:drug effects / Q000502:physiology*
chemical_list: D016222:Fibroblast Growth Factor 2