Pseudohyperphosphorylation has differential effects on polymerization and function of tau isoforms

The microtubule-associated protein tau exists as six isoforms created through the splicing of the second, third, and tenth exons. The isoforms are classified by their number of N-terminal exons (0N, 1N or 2N) and by their number of microtubule-binding repeat regions (3R or 4R). Hyperphosphorylated isoforms accumulate in insoluble aggregates in Alzheimer’s disease and other tauopathies. These neurodegenerative diseases can be categorized based on the isoform content of the aggregates they contain. Hyperphosphorylated tau has the general characteristics of an upward electrophoretic shift, decreased microtubule binding, and an association with aggregation. Previously we have shown that a combination of seven pseudophosphorylation mutations at sites phosphorylated by GSK-3β, referred to as 7-Phos, induced several of these characteristics in full-length 2N4R tau and led to the formation of fewer but longer filaments. We sought to determine whether the same phosphorylation pattern could cause differential effects in the other tau isoforms, possibly through varied conformational effects. Using in vitro techniques, we examined the electrophoretic mobility, aggregation properties and microtubule stabilization of all isoforms and their pseudophosphorylated counterparts. We found that pseudophosphorylation affected each isoform, but in several cases certain isoforms were affected more than others. These results suggest that hyperphosphorylation of tau isoforms could play a major role in determining the isoform composition of tau aggregates in disease

Genotype-Specific Differences between Mouse CNS Stem Cell Lines Expressing Frontotemporal Dementia Mutant or Wild Type Human Tau

Stem cell (SC) lines that capture the genetics of disease susceptibility provide new research tools. To assess the utility of mouse central nervous system (CNS) SC-containing neurosphere cultures for studying heritable neurodegenerative disease, we compared neurosphere cultures from transgenic mice that express human tau with the P301L familial frontotemporal dementia (FTD) mutation, rTg(tauP301L)4510, with those expressing comparable levels of wild type human tau, rTg(tauwt)21221. rTg(tauP301L)4510 mice express the human tauP301L variant in their forebrains and display cellular, histological, biochemical and behavioral abnormalities similar to those in human FTD, including age-dependent differences in tau phosphorylation that distinguish them from rTg(tauwt)21221 mice. We compared FTD-hallmark tau phosphorylation in neurospheres from rTg(tauP301L)4510 mice and from rTg(tauwt)21221 mice. The tau genotype-specific phosphorylation patterns in neurospheres mimicked those seen in mice, validating use of neurosphere cultures as models for studying tau phosphorylation. Genotype-specific tau phosphorylation was observed in 35 independent cell lines from individual fetuses; tau in rTg(tauP301L)4510 cultures was hypophosphorylated in comparison with rTg(tauwt)21221 as was seen in young adult mice. In addition, there were fewer human tau-expressing cells in rTg(tauP301L)4510 than in rTg(tauwt)21221 cultures. Following differentiation, neuronal filopodia-spine density was slightly greater in rTg(tauP301L)4510 than rTg(tauwt)21221 and control cultures. Together with the recapitulation of genotype-specific phosphorylation patterns, the observation that neurosphere lines maintained their cell line-specific-differences and retained SC characteristics over several passages supports the utility of SC cultures as surrogates for analysis of cellular disease mechanisms

Movement distributions of stroke survivors exhibit distinct patterns that evolve with training

BACKGROUND: While clinical assessments provide tools for characterizing abilities in motor-impaired individuals, concerns remain over their repeatability and reliability. Typical robot-assisted training studies focus on repetition of prescribed actions, yet such movement data provides an incomplete account of abnormal patterns of coordination. Recent studies have shown positive effects from self-directed movement, yet such a training paradigm leads to challenges in how to quantify and interpret performance. METHODS: With data from chronic stroke survivors (n = 10, practicing for 3 days), we tabulated histograms of the displacement, velocity, and acceleration for planar motion, and examined whether modeling of distributions could reveal changes in available movement patterns. We contrasted these results with scalar measures of the range of motion. We performed linear discriminant analysis (LDA) classification with selected histogram features to compare predictions versus actual subject identifiers. As a basis of comparison, we also present an age-matched control group of healthy individuals (n = 10, practicing for 1 day). RESULTS: Analysis of range of motion did not show improvement from self-directed movement training for the stroke survivors in this study. However, examination of distributions indicated that increased multivariate normal components were needed to accurately model the patterns of movement after training. Stroke survivors generally exhibited more complex distributions of motor exploration compared to the age-matched control group. Classification using linear discriminant analysis revealed that movement patterns were identifiable by individual. Individuals in the control group were more difficult to identify using classification methods, consistent with the idea that motor deficits contribute significantly to unique movement signatures. CONCLUSIONS: Distribution analysis revealed individual patterns of abnormal coordination in stroke survivors and changes in these patterns with training. These findings were not apparent from scalar metrics that simply summarized properties of motor exploration. Our results suggest new methods for characterizing motor capabilities, and could provide the basis for powerful tools for designing customized therapy

Ombudsman Schemes Energy Sector in Germany, France and the UK

This chapter provides an overview of the most successful ADR model in the EU for resolving consumer disputes: the ombudsman model. After introducing the differences and similarities of both public sector and private sector ombudsman schemes, a comparison of existing consumer ombudsman models in the energy sector in Germany, France, and the UK is offered. Drawing on an extensive empirical dataset of 981 responses to consumer satisfaction surveys of recent users of ADR in the energy sector, cross-national specificities in customer expectations and ombudsman procedures are compared. The ombudsman model is flexible, consumer friendly, and adaptable to changing consumer needs. However, currently no shared language, standards, or training exists in the EU. There is a real opportunity here, if the ADR Directive is used as an incentive to create a harmonized ADR landscape with streamlined procedures and outcomes, consistent terminology, and early management of consumer expectations