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The Journal of Neuroscience, March 21, 2007, 27(12):3260-3267; doi:10.1523/JNEUROSCI.4033-06.2007

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Neurobiology of Disease
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

Kerstin Bendfeldt,¹ Vesna Radojevic,² Josef Kapfhammer,² and Cordula Nitsch¹

¹Section of Neuroanatomy and ²Section of Neurodevelopment, Institute of Anatomy, University of Basel, CH-4056 Basel, Switzerland

Correspondence should be addressed to Dr. Kerstin Bendfeldt at the above address. Email: kerstin.bendfeldt{at}unibas.ch

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.

Key words: blood–brain barrier; FGF-2; occludin; ZO-1; claudin-3; claudin-5

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Received April 15, 2006; revised Feb. 15, 2007; accepted Feb. 18, 2007.

Correspondence should be addressed to Dr. Kerstin Bendfeldt at the above address. Email: kerstin.bendfeldt{at}unibas.ch

This article has been cited by other articles:

				J. A. Perez, S. M. Clinton, C. A. Turner, S. J. Watson, and H. Akil A New Role for FGF2 as an Endogenous Inhibitor of Anxiety J. Neurosci., May 13, 2009; 29(19): 6379 - 6387. [Abstract] [Full Text] [PDF]

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