Talbott John M, Wills Rachel, Shirke Rajendra, Hassanein Leslie, Weinshenker David, Raj Monika
Department of Chemistry, Emory University, Atlanta, Georgia 30322, United States.
Department of Human Genetics, Emory University School of Medicine, Atlanta, Georgia 30322, United States.
JACS Au. 2025 Mar 13;5(4):1717-1727. doi: 10.1021/jacsau.4c01249. eCollection 2025 Apr 28.
Catechol aldehydes (CAs), particularly 3,4-dihydroxyphenylacetaldehyde (DOPAL) and 3,4-dihydroxyphenylglycolaldehyde (DOPEGAL), are potently cytotoxic and have been implicated in pathogenesis of neurodegenerative disorders. Understanding the dynamics of CAs in the brain is crucial for elucidating neurodegenerative pathways. Herein, we present an innovative fluorescent sensor system designed for the selective imaging of CAs within cells and neural tissues. This system employs a dual-reaction trigger, utilizing o-phenylenediamine's selectivity for aldehydes and phenylboronic acid for catechols, generating a specific Förster Resonance Energy Transfer (FRET) signal for CAs. Importantly, we have integrated fluorescence lifetime imaging microscopy (FLIM) with FRET (FLIM-FRET) to enhance detection accuracy while mitigating issues like spectral crosstalk and photobleaching. This dual-reaction FLIM-FRET system allows for the precise visualization of endogenous CAs in the substantia nigra and locus coeruleus of mice, the primary sites of CA production. Notably, this method represents a significant advancement in our ability to study these critical brain regions, as it uniquely enables the tracking of CAs spread across different parts of the brain, addressing a critical gap in the field, as no existing methods allow for such detailed localization of CAs across different brain regions. By enabling precise visualization of CAs within neural tissues, our method enhances understanding of their roles in disease progression.
儿茶酚醛（CAs），特别是3,4 - 二羟基苯乙醛（DOPAL）和3,4 - 二羟基苯乙醇醛（DOPEGAL），具有很强的细胞毒性，并与神经退行性疾病的发病机制有关。了解大脑中CAs的动态对于阐明神经退行性途径至关重要。在此，我们提出了一种创新的荧光传感器系统，用于在细胞和神经组织中对CAs进行选择性成像。该系统采用双反应触发机制，利用邻苯二胺对醛的选择性和苯硼酸对儿茶酚的选择性，产生针对CAs的特定荧光共振能量转移（FRET）信号。重要的是，我们将荧光寿命成像显微镜（FLIM）与FRET（FLIM - FRET）相结合，以提高检测准确性，同时减轻光谱串扰和光漂白等问题。这种双反应FLIM - FRET系统能够精确可视化小鼠黑质和蓝斑中内源性CAs，这是CAs产生的主要部位。值得注意的是，这种方法代表了我们在研究这些关键脑区能力上的重大进步，因为它独特地能够追踪CAs在大脑不同部位的扩散，填补了该领域的一个关键空白，因为现有的方法都无法实现对不同脑区CAs的如此详细定位。通过能够精确可视化神经组织中的CAs，我们的方法增强了对它们在疾病进展中作用的理解。
