Open Access Highly Accessed Research article

Multi-target action of the novel anti-Alzheimer compound CHF5074: in vivo study of long term treatment in Tg2576 mice

Sandra Sivilia1, Luca Lorenzini1, Alessandro Giuliani1, Marco Gusciglio13, Mercedes Fernandez2, Vito Antonio Baldassarro2, Chiara Mangano2, Luca Ferraro34, Vladimiro Pietrini5, Maria Francesca Baroc5, Arturo R Viscomi68, Simone Ottonello6, Gino Villetti7, Bruno P Imbimbo7, Laura Calzà23* and Luciana Giardino23

Author Affiliations

1 Department of Veterinary Medicine, University of Bologna, Bologna, Italy

2 Health Science and Technologies Interdepartmental Center for Industrial Research (HST-ICIR), University of Bologna, Via Tolara di Sopra 50, Bologna, Ozzano Emilia I-40064, Italy

3 IRET Foundation, Via Tolara di Sopra 50, Bologna, Ozzano Emilia 40064, Italy

4 Department of Life Sciences and biotechnology, University of Ferrara, Via Fossato di Mortara 17–19, Ferrara, Italy

5 Department of Neurosciences, Laboratory of Neuropathology, University of Parma, Via Gramsci 14, Parma 43100, Italy

6 Department of Biosciences, Biochemistry and Molecular Biology Unit, Laboratory of Functional Genomics and Protein Engineering, University of Parma, Parma, Italy

7 Research & Development, Chiesi Farmaceutici, Via Palermo 26/A, Parma 43100, Italy

8 Present address: Microbiological Laboratory, GlaxoSmithKline Manufacturing Spa, Via Asolana 90, Parma, S.Polo di Torrile 43056, Italy

For all author emails, please log on.

BMC Neuroscience 2013, 14:44  doi:10.1186/1471-2202-14-44

Published: 5 April 2013

Additional files

Additional file 1: Figure S2:

A, B: Representative images of activated microglia (IBA1 immunostaining) of CA1 hippocampal cortex of Tg2576 vehicle (A) and DAPT (B) treated animals. C, D: Representative images of 6E10 immunostaining of cerebral cortex of Tg2576 vehicle (C) and DAPT (D) treated animals.

Format: PPT Size: 2.2MB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data

Additional file 2: Figure S1:

Representative example of SDS-PAGE fractionation (4-12% Bis-Tris Midi gradient gel) and immunoblot analysis (primary antibody: 6E10 mAb, 1:500; secondary antibody: goat anti-mouse, IrDye 680-labeled antibody, 1:3000) carried out on “low-detergent”, 0.01% NP40/0.1% SDS brain extracts (enriched in extracellular Aβ). Immune-reactive bands were visualized by near-infrared fluorescence (Odyssey imager, LI-COR). Non-specific, 6E10 mAb cross-reactive polypeptides (arrow), present in both wild-type and Tg2576 brain extracts, were used as loading controls and internal references for data normalization. Synthetic prefibrillar Aβ42(n) prepared according to Lambert et al. [32], with n-values ranging from 1 to 4 (not shown), was used as size standard for electrophoretic analysis. Immune-reactive bands were quantified as “near-infrared fluorescence” (NIRF) arbitrary units (see ‘Methods’ for additional details).

Format: PPT Size: 165KB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data

Additional file 3: Figure S3:

Representative images of 6E10 (green) and GFAP- (red) immonostaining in the cerebral cortex of Tg2576 vehicle- (A) and DAPT 375ppm- (B) treated animals. A large number of plaques is observed in both groups, while the GFAP-immunostaining is strongly up-regulated in treated animal. C. semiquantitative evaluation of GFAP immunostaining in the experimental groups. The analysis was performed by evaluating the percentage of plaques surrounded by reactive astrocytes, as evaluated in an area doubling the plaque diameter (see Imbimbo et al.2010, for details). Treated animals show an intense upregulation of GFAP-immunostaining around the plaques. Statisical analysi: one-way ANOVA and Dunnet’s post-hoc test. * p<0.05; **p<0.001.

Format: PDF Size: 3.8MB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data