Comparison of mHTT Antibodies in Huntington’s Disease Mouse Models Reveal Specific Binding Profiles and Steady-State Ubiquitin Levels with Disease Development

<div><p>Huntington’s disease (HD) cellular pathology is characterised by the aggregation of mutant huntingtin (mHTT) protein into inclusion bodies. The present paper compared the sensitivity of five widely used mHTT antibodies (S830; MW8; EM48; 1C2; ubiquitin) against mice from five commonly used HD mouse models (R6/1; YAC128; HdhQ92; B6 HdhQ150; B6 x129/Ola HdhQ150) at two ages to determine: the most sensitive antibodies for each model; whether mHTT antibody binding differed depending on aggregation stage (diffuse versus frank inclusion); the role of ubiquitin during aggregation as the ubiquitin proteosome system has been implicated in disease development. The models demonstrated unique profiles of antibody binding even when the models varied only by background strain (HdhQ150). MW8 was highly sensitive for detecting frank inclusions in all lines whereas EM48, ubiquitin and 1C2 demonstrated consistent staining in all models irrespective of age or form of mHTT. MW8 and S830 were the most sensitive antibodies with 1C2 the least. Ubiquitin levels were stable for each model regardless of age. Ubiquitin was particularly sensitive in young YAC128 mice that demonstrate an absence of inclusions until ~12 months of age suggesting high affinity to mHTT in its diffuse form. The data indicate that generalisations across models regarding the quantification of aggregations may not be valid and that mHTT antibody binding is unique to the mouse model and sensitive to changes in inclusion development.</p></div

High power light microscopy images of mHTT antibody staining of striatal aggregation pathology in early disease HD mice.

<p>The R6/1 mice demonstrated NIIs with all antibodies (A,F,K,P,U), whereas the YAC128 mice demonstrate no NIIs but faint diffuse staining with all antibodies (G,L,Q,V) except S830 (B). The antibodies also failed to detect NIIs in the HdhQ92 line at this age (Photomicrographs C,H,M,R,W) but did demonstrate diffuse staining with all. Both HdhQ150 lines were insensitive to EM48 (N,O) with the original HdhQ150 line also being insensitive to 1C2 (X), but the Photomicrographs for S830 (D), MW8 (I) and ubiquitin (X) in the original line demonstrated good sensitivity for NIIs, as they did for the B6 variant (Photomicrographs E,J, Y respectively). Scale bar = 10 μm.</p

mHTT antibody binding in young and old animals from 5 HD mouse models.

<p>R6/1 mice demonstrate some diffuse staining (A) but high levels of frank inclusions from 4 months of age (B). In this mouse line EM48 was overall the most sensitive antibody, with 1C2 and ubiquitin the least. In young YAC128 mice that demonstrate only diffuse staining (C,D), ubiquitin and S830 were the most effective antibodies. As the mice aged MW8 became the most sensitive antibody. In the HdhQ92 mice, EM48 was the most effective antibody in the young animals (E) with S830 the most effective in the old (F) where the other four antibodies demonstrated a consistent level of mHTT detection. For the HdhQ150 strains, both lines were insensitive to EM48 and demonstrated relatively little binding of diffuse staining per se (original line G,H; B6 backcross I,J) with the antibodies detecting inclusions at high levels even at a young age. The original HdhQ150 line demonstrated consistent levels of detection with the four usable antibodies including 1C2, which was ineffective in the B6 line. MW8 was the most sensitive antibody in the B6 line. Significance markers omitted for clarity (See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155834#pone.0155834.s001" target="_blank">S1 File</a> for raw data).</p

Characteristics of the mouse lines used in the present study.

<p>Characteristics of the mouse lines used in the present study.</p

Means and standard errors of the data sets used in the present study.

<p>(See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155834#pone.0155834.s001" target="_blank">S1 File</a> for raw data).</p

Progetto di recupero e valorizzazione della Cava di Tor Fiscale

Il contributo, corredato dai relativi disegni di progetto, documenta e illustra la soluzione progettuale sperimentale elaborata, come conclusiva della Ricerca Sapienza -finanziata 2012-2013- Responsabile Prof. P.V. Dell'Aira, dal titolo " Sottosuoli urbani La progettazione della 'città che scende'. Tecniche progettuali e realizzative. Identità e qualità spaziale. Comfort ambientale". Il progetto viene presentato attraverso i suoi principali concept informatori: • la relazione tra sopra e sottosuolo • la discenderia, scultura ambientale • le "colonne di luce": landmarks tra sotto e sopra-suolo • il percorso espositivo-narrativo ipogeo • il ridisegno del ponte pedonale • il riuso dell'antico casal