Saltar al contenido
Merck

Integration-independent Transgenic Huntington Disease Fragment Mouse Models Reveal Distinct Phenotypes and Life Span in Vivo.

The Journal of biological chemistry (2015-05-31)
Robert O'Brien, Francesco DeGiacomo, Jennifer Holcomb, Akilah Bonner, Karen L Ring, Ningzhe Zhang, Khan Zafar, Andreas Weiss, Brenda Lager, Birgit Schilling, Bradford W Gibson, Sylvia Chen, Seung Kwak, Lisa M Ellerby
RESUMEN

The cascade of events that lead to cognitive decline, motor deficits, and psychiatric symptoms in patients with Huntington disease (HD) is triggered by a polyglutamine expansion in the N-terminal region of the huntingtin (HTT) protein. A significant mechanism in HD is the generation of mutant HTT fragments, which are generally more toxic than the full-length HTT. The protein fragments observed in human HD tissue and mouse models of HD are formed by proteolysis or aberrant splicing of HTT. To systematically investigate the relative contribution of the various HTT protein proteolysis events observed in vivo, we generated transgenic mouse models of HD representing five distinct proteolysis fragments ending at amino acids 171, 463, 536, 552, and 586 with a polyglutamine length of 148. All lines contain a single integration at the ROSA26 locus, with expression of the fragments driven by the chicken β-actin promoter at nearly identical levels. The transgenic mice N171-Q148 and N552-Q148 display significantly accelerated phenotypes and a shortened life span when compared with N463-Q148, N536-Q148, and N586-Q148 transgenic mice. We hypothesized that the accelerated phenotype was due to altered HTT protein interactions/complexes that accumulate with age. We found evidence for altered HTT complexes in caspase-2 fragment transgenic mice (N552-Q148) and a stronger interaction with the endogenous HTT protein. These findings correlate with an altered HTT molecular complex and distinct proteins in the HTT interactome set identified by mass spectrometry. In particular, we identified HSP90AA1 (HSP86) as a potential modulator of the distinct neurotoxicity of the caspase-2 fragment mice (N552-Q148) when compared with the caspase-6 transgenic mice (N586-Q148).

MATERIALES
Número de producto
Marca
Descripción del producto

Sigma-Aldrich
Agua, Nuclease-Free Water, for Molecular Biology
Sigma-Aldrich
Glicerol, Molecular Biology, ≥99.0%
Sigma-Aldrich
Dodecilsulfatosódico, BioReagent, suitable for electrophoresis, Molecular Biology, ≥98.5% (GC)
Sigma-Aldrich
Agua, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Magnesium chloride solution, Molecular Biology, 1.00 M±0.01 M
Sigma-Aldrich
MOPS, ≥99.5% (titration)
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Triethylamine, ≥99.5%
Sigma-Aldrich
Dodecilsulfatosódico, ≥99.0% (GC), dust-free pellets
Sigma-Aldrich
DL-Ditiotreitol solution, BioUltra, Molecular Biology, ~1 M in H2O
Sigma-Aldrich
Magnesium chloride, ≥98%
Sigma-Aldrich
MES, low moisture content, ≥99% (titration)
Sigma-Aldrich
Dodecilsulfatosódico, ACS reagent, ≥99.0%
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, Molecular Biology, 10% in H2O
Sigma-Aldrich
Digitonina, Used as non-ionic detergent
Supelco
DL-Ditiotreitol solution, 1 M in H2O
Sigma-Aldrich
Triethylamine, ≥99%
Sigma-Aldrich
Dodecilsulfatosódico, ReagentPlus®, ≥98.5% (GC)
Sigma-Aldrich
Glicerol, ≥99.5%
Sigma-Aldrich
Nicotinamide, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
MOPS, BioPerformance Certified, suitable for cell culture, ≥99.5% (titration)
Sigma-Aldrich
Triethylamine, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Glicerol solution, 83.5-89.5% (T)
Sigma-Aldrich
Triethylamine, puriss. p.a., ≥99.5% (GC)
Sigma-Aldrich
Agua, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
Sodium butyrate, 98%
Sigma-Aldrich
Digitonina, ~50% (TLC)
Sigma-Aldrich
Agua, Molecular Biology, sterile filtered
Sigma-Aldrich
Dodecilsulfatosódico, BioUltra, Molecular Biology, ≥99.0% (GC)
Sigma-Aldrich
Glicerol, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for electrophoresis, ≥99% (GC)