Abstract
There is currently no definitive test for early detection of neurodegeneration which is linked with protein aggregation. Finding methods capable of detecting intermediate states of protein aggregates, named oligomers, is critical for the early stage diagnosis of over 30 neurodegenerative diseases including Alzheimer’s or Parkinson’s. Currently, fluorescence-based imaging using Thioflavin T (ThT) dye is the gold standard for detecting protein aggregation. It is used to detect aggregation in vitro and in various tissues, including the cerebrospinal fluid (CSF), whereby the disease-related protein recombinant is seeded with the patient’s fluid. The major drawback of ThT is its lack of sensitivity to oligomeric forms of protein aggregates. Here, we overcome this limitation by transferring a ThT–oligomer mixture into solid state thin films and detecting fluorescence of ThT amplified in the process of stimulated emission. By monitoring the amplified spontaneous emission (ASE) we achieved a remarkable recognition sensitivity to prefibrillar oligomeric forms of insulin and lysozyme aggregates in vitro, to Aβ42 oligomers in the human protein recombinants seeded with CSF and to Aβ42 oligomers doped into brain tissue. Seeding with Alzheimer patient’s CSF containing Aβ42 and Tau aggregates revealed that only Aβ42 oligomers allowed generating ASE. Thus, we demonstrated that, in contrast to the current state-of-the-art, ASE of ThT, a commonly used histological dye, can be used to detect and differentiate amyloid oligomers and evaluate the risk levels of neurodegenerative diseases to potential patients before the clinical symptoms occur.
Highlights
There is currently no definitive test for early detection of neurodegeneration which is linked with protein aggregation
To identify the molecular species formed over time, attenuated total reflectance Fourier-transform infrared spectroscopy (ATRFTIR)[30] and transmission electron microscopy (TEM)[31] were used (Figures 1 and S1)
amplified spontaneous emission (ASE) of Thioflavin T (ThT)-stained proteins has been detected in solid thin films drop-casted from solutions containing amyloids prepared in test tubes, brain tissue doped with amyloid oligomer phantoms, and from human protein recombinants seeded with cerebrospinal fluid (CSF)
Summary
There is currently no definitive test for early detection of neurodegeneration which is linked with protein aggregation. ASE of ThT-stained proteins has been detected in solid thin films drop-casted from solutions containing amyloids prepared in test tubes, brain tissue doped with amyloid oligomer phantoms, and from human protein recombinants seeded with CSF.
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