Author/Authors :
Barbara Baldo، نويسنده , , Paolo Paganetti، نويسنده , , Stephan Grueninger، نويسنده , , David Marcellin، نويسنده , , Linda S. Kaltenbach، نويسنده , , Donald C. Lo، نويسنده , , Martin Semmelroth، نويسنده , , Andjelija Zivanovic، نويسنده , , Dorothee Abramowski، نويسنده , , Donna Smith، نويسنده , , Gregor P. Lotz، نويسنده , , Gillian P. Bates، نويسنده , , Andreas Weiss، نويسنده ,
Abstract :
Huntingtonʹs disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assayʹs efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.
