Abstract
Background Due to the necessity for therapeutic strategies promoting neuro-(re)generation the development of appropriate models appraising the potential of substances for regeneration and repair of neuronal circuits are of great importance. Here we present a procedure for the quantification of fibre outgrowth on the basis of organotypic slice co-cultures of the dopaminergic projection system, appropriate for characterising the effects on axonal growth after treatment with well known controls and growth factors e.g. nerve growth factor (NGF) and new drug candidates. Therefore co-culture preparations were used, consisting of at least two slices of different parts of the brain, namely the ventral tegmental area/substantia nigra (VTA/SN) and the striatum (STR), whereas while incubation fibres projections regenerate and new connections were built, linking the two slices [1,2].
Highlights
Due to the necessity for therapeutic strategies promoting neuro-(re)generation the development of appropriate models appraising the potential of substances for regeneration and repair of neuronal circuits are of great importance
Utilizing immunohistochemistry in combination with laser scanning microscopy the outgrowing fibres in the border region of the ventral tegmental area/substantia nigra (VTA/SN)-STR co-cultures were identified as microtubule associated protein-2 (MAP2), bIII-Tubulin- and tyrosine hydroxylase (TH)-positive
The expression of phosphodiesterase (PDE) - 2A on cell bodies and fibres was revealed within the VTA/SN and STR
Summary
Due to the necessity for therapeutic strategies promoting neuro-(re)generation the development of appropriate models appraising the potential of substances for regeneration and repair of neuronal circuits are of great importance. Results Utilizing immunohistochemistry in combination with laser scanning microscopy the outgrowing fibres in the border region of the VTA/SN-STR co-cultures were identified as microtubule associated protein-2 (MAP2)-, bIII-Tubulin- and tyrosine hydroxylase (TH)-positive. The expression of phosphodiesterase (PDE) - 2A on cell bodies and fibres was revealed within the VTA/SN and STR.
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