Ex vivo culture of isolated zebrafish whole brain

We have succeeded in culturing whole zebrafish brains ex vivo for 1 week. W hile isolated cells and tissue slices have previously been employed for neu robiological studies, these techniques are limited, because while local net works may be preserved, their original context in the whole brain is lost. Culture of the whole brain would facilitate the study of cells and systems within an intact brain infrastructure. Our culture method entailed isolatin g the whole brain and placing it on a sterile and porous membrane, after wh ich it was maintained with a conditioned medium in a six-well plate in a CO , incubator at 28.5 degreesC. Whole brains cultured by this simple method w ere relatively unaltered in terms of their morphology, cytoarchitecture, im munohistochemistry and ability to transport horse radish peroxidase (HRP). This method of cultivation may be very useful for neurobiological research. (C) 2001 Elsevier Science B.V. All rights reserved.