Neuropharmacology

In Vitro Models of Alzheimer’s Disease

Aggregation of amyloid-beta peptides (Aβ) into cytotoxic oligomers and fibrils is one of the major hallmarks of Alzheimer's Disease (AD). Interfering with Aβ aggregation is an inevitable strategy in the development of novel therapeutic approaches. Reliablein vitro models are needed, capable of showing the direct effects of compounds on Aβ oligomer formation to document any beneficial impact these compounds might have on neuron viability.

QPS Neuropharmacology provides an exclusive, fast and reproducible screening assay, Amorfix Aggregated Aβ Assay (A4), able to directly visualize beneficial effects of your developmental compounds on the formation of Aβ aggregates in vitro. As an example, Figure 1 shows the effectiveness of compounds Y and Z in reducing Aβ aggregates, as determined by the A4 assay.

Figure 1: Determination of Aβ aggregates by A4 Assay
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Compounds were co-aggregated with Aβ1-42 for 48h in vitro. Aggregated Aβ was separated from monomers through affinity interaction. After disaggregation, the originally aggregated Aβ was detected using a bead-based immunoassay that applies human Aβ-specific antibody labeled beads and Time Resolved Fluorescence (TRF) measurements. Aβ levels were evaluated as signal to noise ratio (S/N). Co-aggregation of compound Y and Z as well as reference item (RI) with Aβ1-42 reduced aggregate formation. Data are shown as group mean + SEM.

These same compounds were further analyzed for their neuroprotective effects on Aβ-induced toxicity in primary neurons. Indeed, compounds Y and Z, which were able to reduce the formation of Aβ aggregates, also increased cell viability and protected against Aβ-induced toxicity as demonstrated in Figure 2.

Figure 2: Evaluation of neuroprotective effects of compounds on Aβ1-42 induced toxicity
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(A) Compounds were co-aggregated with Aβ1-42 for 48h and applied to primary chicken neurons. After 96h, cell viability was determined according to MTT assay. Co-aggregation of compounds Y and Z as well as RI with Aβ1-42 rescued cell viability and protected against toxic effects of Aβ1-42. Values are expressed as percent of vehicle control. Data are shown as group mean + SEM.

(B) Images illustrate neuroprotective effects of RI co-aggregated with Aβ1-42 (c) compared with cells treated with aggregated Aβ1-42 only (b). RI-treated cells retain the morphological features typical of primary chicken neurons, as compared to control cells (a).

Screen your developmental compounds with our exclusive assay to see whether they reduce Aβ aggregate formation, and are thereby likely to rescue neuronal degeneration in Alzheimer’s Disease.

QPS offers the following in vitro models:
  • H4-APPSL over-expressing cells (A4, Aβ peptides and APP profile, viability)
  • Primary rat/mouse neurons (Aβ toxicity, several other toxicity assays, neurite outgrowth, Aβ peptides and APP profile)
  • SHSY-5Y cells (Aβ toxicity, Tau toxicity, several other toxicity assays, Aβ peptides and APP profile and mitochondrial profile)
  • hTau441 overexpressing SHSY-5Y cell (Tau profile, several toxicity assays, neurite outgrowth, Aβ peptides and APP profile)
  • Primary microglia from rat / mouse and microglial cell lines (inflammation, cytokine profile and oxidative stress)
  • ARPE-19 cells (Aβ toxicity, several other toxicity assays, Aβ peptides and APP profile and mitochondrial profile)
  • Primary human fibroblasts from different patients (Aβ toxicity, several other toxicity assays, Aβ peptides and APP profile and mitochondrial profile)
  • Primary TH-neurons ( 6-OHDa and MPP+ lesions)
  • Alpha synuclein over-expressing RN cells (effects on alpha-synuclein aggregation and several toxicity assays)
  • Customized in vitro models
With various read outs:
  • Screening for secretase inhibitors (determination of Aβ 38, 40, 42 by MSD technology)
  • Screening for effects on APP processing and Aβ peptide formation
  • Tau and activation of tau (several phosphorylation sitesy)
  • Inflammation, cytokine profile and oxidative stress
  • Characterization of cell status (viability, apoptosis and necrosis)
  • Neurite outgrowth (primary rat hippocampal neurons and PC12)
  • Mitochondrial profile (activity, distribution, shape and membrane depolarization)
  • Customized read outs
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