Department of Clinical Neurosciences, Hotchkiss Brain Institute, University of Calgary, Cumming School of Medicine, Calgary, AB, Canada.
Department of Clinical Neurosciences, Hotchkiss Brain Institute, University of Calgary, Cumming School of Medicine, Calgary, AB, Canada.
J Photochem Photobiol B. 2023 Nov;248:112786. doi: 10.1016/j.jphotobiol.2023.112786. Epub 2023 Sep 16.
Protein misfolding with subsequent formation of cross-β-sheet-rich fibrils is a well-known pathological hallmark of various neurodegenerative conditions, including Alzheimer's disease (AD). Recent evidence suggests that specific protein conformations may be the primary drivers of disease progression, differentiation of which remains a challenge with conventional methods. We have previously described a unique phenomenon of light-induced fluorescence enhancement and spectral changes of the amyloid dyes K114 and BSB, and demonstrated its utility in characterizing different amyloid fibrils. In this study, we further characterize and explore the potential of photoconversion, coupled with dual-probe staining, for improved detection of heterogeneity of amyloids using silk fibers and 5xFAD mouse brain sections. BSB and K114 were paired with either Nile Red or MCAAD-3, aiming to increase the sensitivity and specificity of staining and misfolded protein detection via complementary binding and FRET. Principal component analysis of spectral data revealed significant differences between various amyloids, and was able to detect subtle amyloid pathology in the 5xFAD mouse background brain parenchyma.
蛋白质错误折叠，随后形成富含交叉-β-折叠片的纤维，是各种神经退行性疾病（包括阿尔茨海默病（AD））的一个众所周知的病理学特征。最近的证据表明，特定的蛋白质构象可能是疾病进展的主要驱动因素，而传统方法在区分这些构象方面仍然具有挑战性。我们之前描述了一种独特的光诱导荧光增强和淀粉样染料 K114 和 BSB 光谱变化的现象，并证明了其在表征不同淀粉样纤维方面的实用性。在这项研究中，我们进一步对其进行了表征，并探讨了光转化与双探针染色相结合的潜力，以期通过互补结合和 FRET 提高对丝纤维和 5xFAD 小鼠脑切片中淀粉样蛋白异质性的检测灵敏度和特异性。将 BSB 和 K114 与尼罗红或 MCAAD-3 配对，旨在通过互补结合和 FRET 提高染色和错误折叠蛋白检测的灵敏度和特异性。光谱数据的主成分分析显示了各种淀粉样蛋白之间的显著差异，并能够检测到 5xFAD 小鼠脑实质背景中的细微淀粉样病理学。
