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  • Characterization of HTT inclusion size, location, and timing in the zQ175 mouse model of Huntington's disease: an in vivo high-content imaging study.

Characterization of HTT inclusion size, location, and timing in the zQ175 mouse model of Huntington's disease: an in vivo high-content imaging study.

PloS one (2015-04-11)
Nikisha Carty, Nadège Berson, Karsten Tillack, Christina Thiede, Diana Scholz, Karsten Kottig, Yalda Sedaghat, Christina Gabrysiak, George Yohrling, Heinz von der Kammer, Andreas Ebneth, Volker Mack, Ignacio Munoz-Sanjuan, Seung Kwak
RESUMO

Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the huntingtin gene. Major pathological hallmarks of HD include inclusions of mutant huntingtin (mHTT) protein, loss of neurons predominantly in the caudate nucleus, and atrophy of multiple brain regions. However, the early sequence of histological events that manifest in region- and cell-specific manner has not been well characterized. Here we use a high-content histological approach to precisely monitor changes in HTT expression and characterize deposition dynamics of mHTT protein inclusion bodies in the recently characterized zQ175 knock-in mouse line. We carried out an automated multi-parameter quantitative analysis of individual cortical and striatal cells in tissue slices from mice aged 2-12 months and confirmed biochemical reports of an age-associated increase in mHTT inclusions in this model. We also found distinct regional and subregional dynamics for inclusion number, size and distribution with subcellular resolution. We used viral-mediated suppression of total HTT in the striatum of zQ175 mice as an example of a therapeutically-relevant but heterogeneously transducing strategy to demonstrate successful application of this platform to quantitatively assess target engagement and outcome on a cellular basis.

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Sigma-Aldrich
Fluoroshield with DAPI, histology mounting medium
Sigma-Aldrich
Anticorpo antiNeuN (coelho), from rabbit, purified by affinity chromatography
Sigma-Aldrich
Anti-Huntingtin Protein Antibody, clone mEM48, culture supernatant, clone mEM48, Chemicon®