Development and optimization of a rat Purkinje neuron culture to study paraneoplastic cerebellar degeneration

Abstract Background: Elucidation of the mechanisms involved in neurodegenerative disease has been hampered by the lack of robust cellular models that replicate in vivo features. Methods: We developed a refined protocol for generation of age-dependent, well-developed, and synaptically active rat Purkinje neurons in a multilayer cell network culture that can be used to study neurodegenerative diseases such as paraneoplastic cerebellar degeneration (PCD). Result: Independent of the age of the tissue from which the culture was driven, the properties of the generated rat Purkinje neuron culture are complex and robust, allowing great experimental flexibility. Using our model, we found that the application of Yo antibodies, which are associated with PCD, alters the structure of the dendritic arbour of cultured Purkinje neurons within 48h. The numbers of dendrites per branch-order, the branch-order in itself, and the dendritic length were reduced by treatment with anti-Yo. Conclusion: The here presented in vitro model is flexible and can be used to investigate disease mechanisms that disturb Purkinje neuron function and communication in a multilayer network. It proved that anti-Yo has a functional role in the pathogenesis of PCD.