title: TR-FRET-based duplex immunoassay reveals an inverse correlation of soluble and aggregated mutant huntingtin in huntington's disease.
abstract: Huntington's 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 assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.
authors.affiliation: Neuroscience Discovery, Novartis Institutes for BioMedical Research, Basel CH-4002, Switzerland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Baldo
authors.forename: Paolo
authors.initials: P
authors.lastname: Paganetti
authors.forename: Stephan
authors.initials: S
authors.lastname: Grueninger
authors.forename: David
authors.initials: D
authors.lastname: Marcellin
authors.forename: Linda S
authors.initials: LS
authors.lastname: Kaltenbach
authors.forename: Donald C
authors.initials: DC
authors.lastname: Lo
authors.forename: Martin
authors.initials: M
authors.lastname: Semmelroth
authors.forename: Andjelija
authors.initials: A
authors.lastname: Zivanovic
authors.forename: Dorothée
authors.initials: D
authors.lastname: Abramowski
authors.forename: Donna
authors.initials: D
authors.lastname: Smith
authors.forename: Gregor P
authors.initials: GP
authors.lastname: Lotz
authors.forename: Gillian P
authors.initials: GP
authors.lastname: Bates
authors.forename: Andreas
authors.initials: A
authors.lastname: Weiss
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D031541:Fluorescence Resonance Energy Transfer; D055879:Gene Knock-In Techniques; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000378:metabolism* / Q000473:pathology; D007118:Immunoassay; D051379:Mice; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology; D009687:Nuclear Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism
chemical_list: C506812:Htt protein, mouse; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins