Ontology highlight
ABSTRACT: Background
Biochemical analysis of mutant huntingtin (mHTT) aggregation species in HD mice is a common measure to track disease. A longitudinal and systematic study of how tissue processing affects detection of conformers has not yet been reported. Understanding the homeostatic flux of mHTT over time and under different processing conditions would aid in interpretation of pre-clinical assessments of disease interventions.Objective
Provide a systematic evaluation of tissue lysis methods and molecular and biochemical assays in parallel with behavioral readouts in R6/2 mice to establish a baseline for HTT exon1 protein accumulation.Methods
Established biochemical methods were used to process tissue from R6/2 mice of specific ages following behavior tasks. Aggregation states and accumulation of mHTT exon 1 protein were evaluated using multiple break and assay methods to determine potential conformational flux assay specificity in detection of mHTT species, and tissue specificity of conformers.Results
Detection of mHTT exon 1 protein species varied based on biochemical processing and analysis providing a baseline for subsequent studies in R6/2 mice. Insoluble, high molecular weight species of mHTT exon 1 protein increased and tracked with onset of behavioral impairments in R6/2 mice using multiple assay methods.Conclusions
Conformational flux from soluble monomer to high molecular weight, insoluble species of mHTT exon 1 protein was generally consistent for multiple assay methods throughout R6/2 disease progression; however, the results support the use of multiple biochemical techniques to detect mHTT exon 1 protein species for preclinical assessments in HD mouse models expressing mHTT exon 1 protein.
SUBMITTER: Morozko EL
PROVIDER: S-EPMC6294605 | biostudies-literature | 2018
REPOSITORIES: biostudies-literature
Morozko Eva L EL Ochaba Joseph J Hernandez Sarah J SJ Lau Alice A Sanchez Isabella I Orellana Iliana I Kopan Lexi L Crapser Joshua J Duong Janet H JH Overman Julia J Yeung Silvia S Steffan Joan S JS Reidling Jack J Thompson Leslie M LM
Journal of Huntington's disease 20180101 4
<h4>Background</h4>Biochemical analysis of mutant huntingtin (mHTT) aggregation species in HD mice is a common measure to track disease. A longitudinal and systematic study of how tissue processing affects detection of conformers has not yet been reported. Understanding the homeostatic flux of mHTT over time and under different processing conditions would aid in interpretation of pre-clinical assessments of disease interventions.<h4>Objective</h4>Provide a systematic evaluation of tissue lysis m ...[more]