Pular para o conteúdo
Merck
  • Use of adeno-associated virus-mediated delivery of mutant huntingtin to study the spreading capacity of the protein in mice and non-human primates.

Use of adeno-associated virus-mediated delivery of mutant huntingtin to study the spreading capacity of the protein in mice and non-human primates.

Neurobiology of disease (2020-05-23)
Alexander Maxan, Giacomo Sciacca, Melanie Alpaugh, Zhu Tao, Ludivine Breger, Benjamin Dehay, Zhang Ling, Qin Chuan, Giulia Cisbani, Maria Masnata, Shireen Salem, Steve Lacroix, Abid Oueslati, Erwan Bezard, Francesca Cicchetti
RESUMO

In order to model various aspects of Huntington's disease (HD) pathology, in particular protein spread, we administered adeno-associated virus (AAV) expressing green fluorescent protein (GFP) or GFP coupled to HTT-Exon1 (19Q or 103Q) to the central nervous system of adult wild-type (WT) mice and non-human primates. All animals underwent behavioral testing and post-mortem analyses to determine the long-term consequences of AAV injection. Both mice and non-human primates demonstrated behavioral changes at 2-3 weeks post-surgery. In mice, these changes were absent after 3 months while in non-human primates, they persisted in the majority of tested animals. Post-mortem analysis revealed that spreading of the aggregates was limited, although the virus did spread between synaptically-connected brain regions. Despite circumscribed spreading, the presence of mHTT generated changes in endogenous huntingtin (HTT) levels in both models. Together, these results suggest that viral expression of mHTTExon1 can induce spreading and seeding of HTT in both mice and non-human primates.

MATERIAIS
Número do produto
Marca
Descrição do produto

Sigma-Aldrich
Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Sigma-Aldrich
Glicerol, for molecular biology, ≥99.0%
Sigma-Aldrich
Soro bovino fetal, USA origin, sterile-filtered, suitable for cell culture, suitable for hybridoma
Sigma-Aldrich
Dodecilsulfato de sódio, BioReagent, suitable for electrophoresis, for molecular biology, ≥98.5% (GC)
Sigma-Aldrich
Solução antimicótica antibiótica (100×), estabilizada, with 10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Bissulfito de sódio, ACS reagent
Sigma-Aldrich
2-Mercaptoetanol, for molecular biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
Cloridrato de Trizma®, reagent grade, ≥99.0% (titration), crystalline
Sigma-Aldrich
Gelatin solution, Type B, 2% in H2O, tissue culture grade, BioReagent, suitable for cell culture
Sigma-Aldrich
Soro asinino
Sigma-Aldrich
BIS-TRIS, ≥98.0% (titration)
Sigma-Aldrich
Evans Blue, Dye content ≥75 %
Sigma-Aldrich
Poli-D-lisina, mol wt 70,000-150,000
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
Fosfato de sódio, ReagentPlus®, ≥99.0%
Sigma-Aldrich
BICINE, BioXtra, ≥99% (titration)
Sigma-Aldrich
Fosfato de sódio, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Tricina, BioPerformance Certified, suitable for cell culture, ≥99% (titration)
Sigma-Aldrich
Bromophenol Blue, titration: suitable
SAFC
Meio de Eagle modificado por Dulbecco/Mistura de nutrientes de Ham F-12, liquid, suitable for cell culture