Aslund Andreas, Sigurdson Christina J, Klingstedt Therése, Grathwohl Stefan, Bolmont Tristan, Dickstein Dara L, Glimsdal Eirik, Prokop Stefan, Lindgren Mikael, Konradsson Peter, Holtzman David M, Hof Patrick R, Heppner Frank L, Gandy Samuel, Jucker Mathias, Aguzzi Adriano, Hammarström Per, Nilsson K Peter R
Department of Chemistry, IFM, Linköping University, Sweden.
ACS Chem Biol. 2009 Aug 21;4(8):673-84. doi: 10.1021/cb900112v.
Molecular probes for selective identification of protein aggregates are important to advance our understanding of the molecular pathogenesis underlying cerebral amyloidoses. Here we report the chemical design of pentameric thiophene derivatives, denoted luminescent conjugated oligothiophenes (LCOs), which could be used for real-time visualization of cerebral protein aggregates in transgenic mouse models of neurodegenerative diseases by multiphoton microscopy. One of the LCOs, p-FTAA, could be utilized for ex vivo spectral assignment of distinct prion deposits from two mouse-adapted prion strains. p-FTAA also revealed staining of transient soluble pre-fibrillar non-thioflavinophilic Abeta-assemblies during in vitro fibrillation of Abeta peptides. In brain tissue samples, Abeta deposits and neurofibrillary tangles (NFTs) were readily identified by a strong fluorescence from p-FTAA and the LCO staining showed complete co-localization with conventional antibodies (6E10 and AT8). In addition, a patchy islet-like staining of individual Abeta plaque was unveiled by the anti-oligomer A11 antibody during co-staining with p-FTAA. The major hallmarks of Alzheimer's disease, namely, Abeta aggregates versus NFTs, could also be distinguished because of distinct emission spectra from p-FTAA. Overall, we demonstrate that LCOs can be utilized as powerful practical research tools for studying protein aggregation diseases and facilitate the study of amyloid origin, evolution and maturation, Abeta-tau interactions, and pathogenesis both ex vivo and in vivo.
用于选择性识别蛋白质聚集体的分子探针对于增进我们对脑淀粉样变性潜在分子发病机制的理解非常重要。在此，我们报告了五聚噻吩衍生物的化学设计，即发光共轭低聚噻吩（LCOs），其可通过多光子显微镜用于神经退行性疾病转基因小鼠模型中脑蛋白质聚集体的实时可视化。其中一种LCO，p - FTAA，可用于对两种小鼠适应朊病毒株的不同朊病毒沉积物进行离体光谱分析。p - FTAA还揭示了在β - 淀粉样肽（Abeta）体外纤维化过程中短暂可溶性前纤维状非硫黄素亲和性Abeta聚集体的染色情况。在脑组织样本中，p - FTAA发出的强烈荧光很容易识别出Abeta沉积物和神经原纤维缠结（NFTs），并且LCO染色显示与传统抗体（6E10和AT8）完全共定位。此外，在与p - FTAA共染色过程中，抗寡聚体A11抗体揭示了单个Abeta斑块的斑片状胰岛样染色。由于p - FTAA有不同的发射光谱，阿尔茨海默病的主要特征，即Abeta聚集体与NFTs，也能够被区分开来。总体而言，我们证明LCOs可作为研究蛋白质聚集疾病的强大实用研究工具，并有助于研究淀粉样蛋白的起源、进化和成熟、Abeta - tau相互作用以及离体和体内的发病机制。
