Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment
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...
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doaj-d3e563eb8d99412b957653a25b2294ea2021-03-20T04:51:31ZengElsevierNeurobiology of Disease1095-953X2005-12-01203785798Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragmentDiana Zala0Alexandra Benchoua1Emmanuel Brouillet2Valérie Perrin3Marie-Claude Gaillard4Anne D. Zurn5Patrick Aebischer6Nicole Déglon7Institute of Neurosciences, Swiss Federal Institute of Technology Lausanne, EPFL, 1015 Lausanne, SwitzerlandURA2210, CEA/CNRS, 91401 Orsay, FranceURA2210, CEA/CNRS, 91401 Orsay, FranceInstitute of Neurosciences, Swiss Federal Institute of Technology Lausanne, EPFL, 1015 Lausanne, SwitzerlandService de Biochimie et de Génétique Moléculaire, Département de Biologie Joliot-Curie, Commissariat à l’Energie Atomique (CEA) Saclay, 91191 Gif-sur-Yvette Cedex, FranceDepartment of Experimental Surgery, University Medical School Lausanne, 1011 Lausanne, SwitzerlandInstitute of Neurosciences, Swiss Federal Institute of Technology Lausanne, EPFL, 1015 Lausanne, SwitzerlandInstitute of Neurosciences, Swiss Federal Institute of Technology Lausanne, EPFL, 1015 Lausanne, Switzerland; Department of Medical Research, ImaGene Program, CEA, 91401 Orsay, France; Corresponding author. Commissariat à l’Energie Atomique (CEA), Service Hospitalier Frédéric Joliot, 4, Place du Général Leclerc, 91401 Orsay, Cedex, France. Fax: +33 1 69 86 77 45.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.http://www.sciencedirect.com/science/article/pii/S096999610500149XStriatal degenerationLentiviral vectorMutant huntingtin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Diana Zala Alexandra Benchoua Emmanuel Brouillet Valérie Perrin Marie-Claude Gaillard Anne D. Zurn Patrick Aebischer Nicole Déglon |
spellingShingle |
Diana Zala Alexandra Benchoua Emmanuel Brouillet Valérie Perrin Marie-Claude Gaillard Anne D. Zurn Patrick Aebischer Nicole Déglon Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment Neurobiology of Disease Striatal degeneration Lentiviral vector Mutant huntingtin |
author_facet |
Diana Zala Alexandra Benchoua Emmanuel Brouillet Valérie Perrin Marie-Claude Gaillard Anne D. Zurn Patrick Aebischer Nicole Déglon |
author_sort |
Diana Zala |
title |
Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment |
title_short |
Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment |
title_full |
Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment |
title_fullStr |
Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment |
title_full_unstemmed |
Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment |
title_sort |
progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment |
publisher |
Elsevier |
series |
Neurobiology of Disease |
issn |
1095-953X |
publishDate |
2005-12-01 |
description |
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. |
topic |
Striatal degeneration Lentiviral vector Mutant huntingtin |
url |
http://www.sciencedirect.com/science/article/pii/S096999610500149X |
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