Gadolinium-complexed Aβ-binding contrast agents for MRI diagnosis of Alzheimer's Disease  Balpreet Matharu, Nick Spencer, Franklyn Howe, Brian Austen  Neuropeptides  Volume 53, Pages 63-70 (October 2015) DOI: 10.1016/j.npep.2015.07.001 Copyright © 2015 Elsevier Ltd Terms and Conditions

Fig. 1 Binding of biotinylated imaging reagents R1–R5 to coated films in ELISA plates of Αβ40 in oligomeric (o) or fibrillar (f) forms, performed as described (Matharu et al, 2010). Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions

Fig. 2 Surface plasmon resonance response units (RU) (y-axis) with baseline taken as zero, against time in secs (x-axis) of 100μM solutions of R1, R2, R3 or R4 injected over Aβ40 immobilised on a T100 carboxylate surface. Response to a control surface with bound non-specific protein was subtracted. Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions

Fig. 3 Brain sections (10μm) cut from post-mortem brain samples either from an AD patient (A, B and D) or young control (C) was stained with biotinylated-R2 (A, B & C), or the 6E10 antibody to Aβ (Signet) (D). In the micrograph (right side), SW-APP-transfected 293 cells were treated in culture for 2h with proteasome inhibitor lactacystin (10μM) to accumulate Αβ, then with biotinylated-R2 (10μΜ) for 1h, harvested in chamber slides (Thermo) and stained with TRITC-avidin (Sigma), to give intracellular red fluorescence of the imaging agent binding to aggregated Aβ under a microscope (×40 magnification) by viewing through the rhodamine filter. Appropriate controls did not show red fluorescence. Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions

Fig. 4 Ex vivo MRI imaging of reagents. 60μm sections were cut from 2month-old 5XFAD mice in which histological staining had shown hippocampal β-amyloid deposition (Spencer et al, 2013) and from B65JF1/J non-transgenic control mice of the same age. Sections were incubated with 0.5ml of 2mM reagents R1, R2 and R3 in PBS, and then washed with PBS prior to sealing with glue in a slide with cover well slip together with Flourinert FC-77 (3M Corporation, USA). T1 weighted MRI (TR/TE/flip of 30ms/4.7ms/15° was acquired with 0.16mm in plane resolution after placing on a small imaging coil. The intensities shown are the effectiveness of reagents to reduce T1 and so increase intensity. Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions

Fig. 5 Average biodistribution of 153Gd(DOTA)GrffvlkKrrrrrr-NH2(R2) in CD-1 mice in all organs. Shown are the %ID amounts biodistributed into organs, tissues and blood dissected at increasing time points in three mice at each time point. Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions

Fig. 6 In vivo MRI scanning up to 80min after FUS activation showed a specific in vivo increase in MRI T1 response achieved by the reagent R2 (Rnob) across the cortex and thalamus scan at T=0 is shown upper left, and scan at T=10min at lower left. Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions

Neuropeptides 2015 53, 63-70DOI: (10.1016/j.npep.2015.07.001) Copyright © 2015 Elsevier Ltd Terms and Conditions