title: Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment.
abstract: A lentiviral vector expressing a mutant huntingtin protein (htt171-82Q) was used to generate a chronic model of Huntington's disease (HD) in rat primary striatal cultures. In this model, the majority of neurons expressed the transgene so that Western blot analysis and flow cytometry measurement could complement immunohistological evaluation. Mutant huntingtin produced a slowly progressing pathology characterized after 1 month by the appearance of neuritic aggregates followed by intranuclear inclusions, morphological anomalies of neurites, loss of neurofilament 160, increased expression in stress response protein Hsp70, and later loss of neuronal markers such as NeuN and MAP-2. At 2 months post-infection, a significant increase in TUNEL-positive cells confirmed actual striatal cell loss. Interestingly, cortical cultures infected with the same vector showed no sign of neuronal dysfunction despite accumulation of numerous inclusions. We finally examined whether the trophic factors CNTF and BDNF that were found neuroprotective in acute HD models could prevent striatal degeneration in a chronic model. Results demonstrated that both agents were neuroprotective without modifying inclusion formation. The present study demonstrates that viral vectors coding for mutant htt provides an advantageous system for histological and biochemical analysis of HD pathogenesis in primary striatal cultures.
authors: Institute of Neurosciences, Swiss Federal Institute of Technology Lausanne, EPFL, 1015 Lausanne, Switzerland.
authors: Diana
authors: D
authors: Zala
authors: Alexandra
authors: A
authors: Benchoua
authors: Emmanuel
authors: E
authors: Brouillet
authors: Valérie
authors: V
authors: Perrin
authors: Marie-Claude
authors: MC
authors: Gaillard
authors: Anne D
authors: AD
authors: Zurn
authors: Patrick
authors: P
authors: Aebischer
authors: Nicole
authors: N
authors: Déglon
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology / Q000627:therapeutic use; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D020934:Ciliary Neurotrophic Factor / Q000494:pharmacology / Q000627:therapeutic use; D003342:Corpus Striatum / Q000378:metabolism* / Q000503:physiopathology* / Q000821:virology; D004268:DNA-Binding Proteins; D005822:Genetic Vectors / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D002479:Inclusion Bodies / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016086:Lentivirus / Q000235:genetics; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009410:Nerve Degeneration / Q000235:genetics* / Q000378:metabolism / Q000517:prevention & control; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods; D019076:Transgenes / Q000235:genetics
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; D004268:DNA-Binding Proteins; D018840:HSP70 Heat-Shock Proteins; C086055:HTT protein, human; D000071058:Huntingtin Protein; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D016900:Neurofilament Proteins; D009687:Nuclear Proteins; C061195:neurofilament protein M