Development of Fluorescent Methods for the Detection of Amyloidosis in Neurodegeneration
At the molecular level, neurodegeneration is characterized by the appearance of aggregates of misfolded proteins known as amyloids. These assemblies are often associated with neurotoxicity and are classically the defining feature of neurodegenerative diseases such as Alzheimer’s Disease (AD). The appearance of these aggregates in the brain often preludes the clinical symptoms of neurodegeneration, and thus are valuable targets for the early diagnoses of the diseases.
This thesis documents the development of amino-naphthalene cyanoacrylate (ANCA) fluorophores and their use towards detecting these amyloid aggregates. Chapters 2 and 3 describe the rational design and synthesis of the ANCA family and their colorimetric sensitivity to different amyloid species. This unique capability was found to originate from their environmentally sensitive molecular structure, which was utilized to characterize amyloids based on their fluorescence emission lambda max.
Chapter 4 describes the development of an in vivo imaging methodology to detect amyloidosis in the retina of AD and prion mice using the fluorescence of an ANCA derivative, ARCAM-1. This preliminary study on retinal imaging supports the use of the eye as a window to peer into the central nervous system.
Finally, Chapter 5 presents the utility of ARCAM-1 as a sensor for the detection of the smaller, transient oligomeric amyloid species by use of fluorescence correlation spectroscopy (FCS), and as a sensor for amyloidosis in the urine of pregnant women with pre-eclampsia (PE).