Alzheimer's Disease tissue culture models
JSW provides different cell culture models for Alzheimer's Disease including induced neuronal cell death, analyses of human APP cleavage and Tau phosphorylation.
1) In vitro lesion models
It is meanwhile well described, that neuronal loss in case of AD is due to various types of stress conditions. These comprise oxidative stress, excitotoxicity, Ca++-overload and others beyond amyloid toxicity.
To induce suchlike conditions embryonic neuronal cultures and neuron/glia co-cultures are treated with agents that induce lesions mimicking important aspects of Alzheimer's Disease:
Cultures are analysed for cell survival, apoptotic and necrotic phenotypes and expression of different synaptic markers (see Analyses of culture models).
2) Screening of secretase inhibitors
Generation of Aß1-42 species is a hallmark of Alzheimer's Disease. This assay allows the identification of compounds that interfere with the cleavage of human APP. We provide three different cell culture models to analyse the effect of pharmacological agents on generation of different Aß and sAPP peptides. In all assays, Aß peptides in supernatants are quantified by an immunosorbent assay.
- Primary chicken telencephalic neurons. These neurons secrete endogenous wildtype Aß peptides including Aß38, Aß40 and Aß42. Both human and chick Aß42 are identical in sequence. Therefore, this model addresses processing of endogenous wildtype APP in primary neurons.
- Primary rat hippocampal neurons. These neurons are phenotypically closer to adult neurons than neurons cultured for shorter periods as they have extended a dense network of neuronal processes and are likely to possess already intrinsic synaptic activity.
- H4 neuroglioma cells overexpressing human APP containing the Swedish double mutation. These cells enable to analyze compounds for their effect on precessing of human mutant APP.
Here, see also our new publication for reference:
Comparison of Pharmacological Modulation of APP Metabolism in Primary Chicken Telencephalic Neurons and in a Human Neuroglioma Cell Line
Stefan Czvitkovich; Stephan Duller; Else Mathiesen; Klaus Lorenzoni; Bruno P. Imbimbo; Birgit Hutter-Paier; Manfred Windisch, Robert Wronski
3) Analyses of Tau phosphorylation
Increased Tau phosphorylation and the formation of neurofibrillary tangles are critical appearances of Alzheimer's Disease. We have two cell culture systems that allow the analysis of compounds that interfere with TAU phosphorylation
- SH-SY5Y cells overexpressing human mutant TAU441 harboring two disease related mutations (V337M/R406W). These cells highly overexpress hyperphosphorylated TAU including multiple phosphoepitopes linked with AD.
- Primary cultures of rat hippocampal neurons. Primary neurons are treated with okadaic acid, a known phosphatase 1 inhibitor.
These cultures can be examined by ELISA, Western Blotting, or Mass Spectrometry for the following pTAU epitopes:
- Ser396/404
- Ser202/Thr205
- Thr231
- Thr181
- and others
