The fluorescent pentameric oligothiophene pFTAA identifies filamentous tau in live neurons cultured from adult P301S tau mice

Frontiers in Neuroscience

Peter eNilsson¹  Michel eGoedert²  Therese eKlingstedt²  Aviva eTolkovsky³  Jack eBrelstaff³  Maria Grazia eSpillantini³  Bernardino eOssola³  Jonas eNeher⁴ 

[1] Linkoping University;MRC Laboratory of Molecular Biology;University of Cambridge;University of Tubingen;

关键词: Neurofibrillary Tangles;    Alzheimer’s disease;    Frontotemporal dementia (FTD);    Hyperphosphorylated tau;    Dorsal root ganglion neurons;    transgenic P301S tau mouse;   

DOI  :  10.3389/fnins.2015.00184

来源: DOAJ

【 摘 要 】

Identification of fluorescent dyes that label filamentous protein aggregates characteristic of neurodegenerative disease, such as β-amyloid and tau in Alzheimer’s disease (AD), in a live cell culture system has previously been a major hurdle. Here we show that pentameric formyl thiophene acetic acid (pFTAA) fulfils this function in living neurons cultured from adult P301S tau transgenic mice. Injection of pFTAA into 5-month-old P301S tau mice detected cortical and DRG neurons immunoreactive for AT100, which identifies solely filamentous tau, or MC1, which identifies a conformational change in tau that is commensurate with neurofibrillary tangle formation in AD brains. In fixed cultures of dorsal root ganglion (DRG) neurons, pFTAA binding, which also identified AT100 or MC1+ve neurons, followed a single, saturable binding curve with a half saturation constant of 0.14 µM, the first reported measurement of a binding affinity of a beta-sheet reactive dye to primary neurons harbouring filamentous tau. Treatment with formic acid, which solubilises filamentous tau, extracted pFTAA, and prevented the re-binding of pFTAA and MC1 without perturbing expression of soluble tau, detected using an anti-human tau (HT7) antibody. In live cultures, pFTAA only identified DRG neurons that, after fixation, were AT100/MC1+ve, confirming that these forms of tau pre-exist in live neurons. The utility of pFTAA to discriminate between living neurons containing filamentous tau from other neurons is demonstrated by showing that more pFTAA+ve neurons die than pFTAA-ve neurons over 25 days. Since pFTAA identifies fibrillar tau and other misfolded proteins in living neurons in culture, in animal models of several neurodegenerative diseases, and in human brains, it should have considerable application in sorting out disease mechanisms and in identifying disease-modifying drugs that will ultimately help establish the mechanisms of neurodegeneration in human neurodegenerative diseases.

【 授权许可】

Unknown