CrossSeeds

About

The JPND CrossSeeds project hypothesis is that AD, PD and HD share common pathways of protein aggregation and that cross-seeding of pathogenic pGlu-modified proteins are leading to neurodegeneration. This site demonstrates ongoing efforts in mapping of the distribution of pGlu-protein expression sites in rat and mouse models. High resolution histological section images from whole brains are organized in a database and anchored to 3D reference atlas space (Waxholm rat brain atlas and Allen mouse brain mouse atlas). The data system in preparation allows storage of all raw data, data retrieval, and visualization and analysis in 3D space.

Contact info: Maja A. Puchades, Nesys laboratory, University of Oslo, Norway. Email: m.a.puchades@medisin.uio.no

Image repository, viewing, and analysis

The virtual microscopy filmstrip viewer allows interactive zooming and panning of images.

The Localizoom viewer uses the atlas anchoring information and allows visualization of single section images with a transparent atlas-segmentation overlay, matching the location in the brain of the section image and cut at the custom angle of orientation of the histological section. Names of atlas structures are shown when pointing the mouse over the region of interest and 3D coordinates at the location of the cursor are shown real-time. An extended version of the LocaliZoom can harvest coordinates corresponding to selected features of interest in the images for further 3D analysis. Zooming in a specific area is possible using the mouse wheel. Transparency of the atlas is adjustable on the top left corner scale.

AD mouse tg2576

Two 18 month old Alzheimer’s mice (Mtg01_bl1 and m-287) were labelled with 4 different antibodies: against human APP (1D1) (Höfling et al. 2016 Aging Cell), against Aβ peptide (aa 17-24) (BioLegend,Cat:4G8), against Aβ peptide (aa 1-4) but not APP (Cayman Chemicals, Cat:MOAB-2) and against Pyroglutamate-modified Aβ peptides (Aβ_pE3) (pGlu)(Probiodrug). Images of sections labelled with each of the antibodies can be viewed with the filmstrip viewer link. The atlas overlayed sections are available using the LocaliZoom tool as outlined above.

One tg2576 female mouse (m-543;18 months) was labelled by triple immunofluorescence labelling with antibodies against Aβ_pE3 (J8) conj. with Cy2 (green); α-synuclein (C20R) conj. with Cy3 (orange); Aβ peptide (aa 17-24) (4G8) conjugated with Cy5 (red) and Hoest labelling of Nuclei (Dapi channel)(blue). The triple labelling shows localization of α-synuclein close to amyloid plaques, thereby comforting the CrossSeeding hypothesis.

Human APP (1D1)

Mouse Mtg01_bl1 (Female): filmstrip Localizoom (Ctrl+click to open in a new window)

Mouse m-287 (Male): filmstrip Localizoom

Aβ peptide (aa 17-24)(4G8)

Mouse Mtg01_bl1 (Female): filmstrip Localizoom

Mouse m-287 (Male): filmstrip Localizoom

Aβ peptide (aa 1-4) MOAB-2

Mouse Mtg01_bl1 (Female): filmstrip Localizoom

Mouse m-287 (Male): filmstrip Localizoom

Aβ_pE3 (pGlu)

Mouse Mtg01_bl1 (Female): filmstrip Localizoom

Mouse m-287 (Male): filmstrip Localizoom

Triple labelling (4G8; C20R and pGlu)

Mouse m-543_bl1 (Female): filmstrip

AD rat (McGill-R-Thy1-APP)

This 16 month old male rat was labelled with with 4 different antibodies: against human APP (1D1) (Höfling et al. 2016 Aging Cell), against Aβ peptide aa 17-24 (BioLegend,Cat:4G8), against Aβ peptide (aa 1-4) but not APP (Cayman Chemicals, Cat:MOAB-2) and against Pyroglutamate-modified Aβ peptides (Aβ_pE3) (pGlu)(Probiodrug). Images of sections labelled with each of the antibodies can be viewed with the filmstrip viewer link. The atlas overlayed sections are available using the LocaliZoom tool as outlined above.

Human APP (1D1)

Rat r4753 (Male): filmstrip Localizoom (Ctrl+click to open in a new window)

PD mouse (TetO-A53T-PITX3)

A 6 month old mouse was labelled with 3 different antibodies: against human α-synuclein (aa 117-131) (BML-SA3400) (Enzo Lifesciences), against mouse α-synuclein (D37A6) (Cell Signalling) and against Phospho-α-synuclein (Ser129) (EP1536Y)(Abcam, Cat:ab51253).

Human α-synuclein (aa 117-131)

Mouse M9 : filmstrip Localizoom (Ctrl+click to open in a new window)

Mouse α-synuclein (D37A6)

Mouse M9 : filmstrip Localizoom

Phospho-α-synuclein (Ser129)

Mouse M9 : filmstrip Localizoom

PD rat (BAC-SNCA)

Two transgenic male Parkinson rats (Tg5573; 15 month oldand Tg 5141; 12 months old) and two wild type rats (WT 5518 and WT5559;15 months old) were labelled with antibodies against α-synuclein: against α-synuclein Syn1 (endogenous) (BD, Cat: 610787), against aggregated α-synuclein (Millipore, Cat.: MABN389), against alpha Synuclein 15G7 (human) (Enzo, Cat.: ALX-804-258). In addition, these animals were labelled with antibodies against Aβ peptide aa 17-24 (BioLegend, Cat:4G8), against pyroGlu-alpha Synuclein (pGlu79) (Probiodrug) and against Huntingtin (Epitomics/Abcam, Cat: ab45169).