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A novel ex vivo Huntington’s disease model for studying GABAergic neurons and cell grafts by laser microdissection

Abstract : Organotypic brain slice cultures have been recently used to study neurodegenerative disorders such as Parkinson's disease and Huntington's disease (HD). They preserve brain three-dimensional architecture, synaptic connectivity and brain cells microenvironment. Here, we developed an innovative model of Huntington's disease from coronal rat brain slices, that include all the areas involved in the pathology. HD-like neurodegeneration was obtained in only one week, in a single step, during organotypic slice preparation, without the use of neurotoxins. HD-like histopathology was analysed and after one week, a reduction of 40% of medium spiny neurons was observed. To analyse new therapeutic approaches in this innovative HD model, we developed a novel protocol of laser microdissection to isolate and analyse by RT-qPCR, grafted cells as well as surrounding tissue of fresh organotypic slices. We determined that laser microdissection could be performed on a 400μm organoty-pic slice after alcohol dehydration protocol, allowing the analysis of mRNA expression in the rat tissue as well as in grafted cells. In conclusion, we developed a new approach for modeling Huntington's disease ex vivo, and provided a useful innovative method for screening new potential therapies for neurodegenerative diseases especially when associated with laser microdissection.
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Submitted on : Monday, June 11, 2018 - 4:21:48 PM
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E M André, N. Daviaud, Laurence Sindji, J. Cayon, R. Perrot, et al.. A novel ex vivo Huntington’s disease model for studying GABAergic neurons and cell grafts by laser microdissection. PLoS ONE, Public Library of Science, 2018, 13 (3), pp.e0193409. ⟨10.1371/journal.pone.0193409⟩. ⟨inserm-01812641⟩



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