Organotypic vibrosections: novel whole sagittal brain cultures

In vitro cell culture models are of enormous importance in neuroscience research and organotypic brain slices are found to be a potent model very close to the in vivo situation. Brain slices can be cultured as single slices or as co-slices. However, there is need to culture whole brain sections, containing the complex functional architecture. The aim of the present study was to develop and characterize whole brain sagittal slice cultures (200μm organotypic vibrosections) from postnatal day 8 rats. We show that sagittal vibrosections can be cultured for several weeks and they maintain survival of cholinergic and dopaminergic neurons, as well as a strong capillary network. Partly long-distance cortico-striatal and cortico-hippocampal nerve fibers were found using Mini-Ruby neurotracing. Dopaminergic nerve fibers extended from the mesencephalon, but in the striato-nigral tract and in the striatum only strong dense varicosities were found. The model also allows to study pathological triggers, such as e.g. hydrogen peroxide markedly increased propidiumiodide-positive nuclei in the hippocampus. In conclusion, our novel model provides an easy potent whole sagittal brain culture system that allows to study cholinergic and dopaminergic neurons together but also in close interaction with all other cells of the brain and with capillaries. It will be a great challenge in future to use this model to re-construct whole pathways. This vibrosection model may partly represent a close adult in vivo situation, which allows to study neurodegeneration and neuroprotection of cholinergic and dopaminergic neurons, which plays an important role in Alzheimer's and Parkinson's disease, respectively.