Mitochondrial superoxide (O2⋅−) underlies much oxidative damage and redox signaling. Fluorescent probes can detect O2⋅−, but are of limited applicabilityin vivo, while in cells their usefulness is constrained by side reactions and DNA intercalation. To overcome these limitations, we developed a dual-purpose mitochondrial O2⋅−probe, MitoNeoD, which can assess O2⋅−changesin vivoby mass spectrometry andin vitroby fluorescence. MitoNeoD comprises a O2⋅−-sensitive reduced phenanthridinium moiety modified to prevent DNA intercalation, as well as a carbon-deuterium bond to enhance its selectivity for O2⋅−over non-specific oxidation, and a triphenylphosphonium lipophilic cation moiety leading to the rapid accumulation within mitochondria. We demonstrated that MitoNeoD was a versatile and robust probe to assess changes in mitochondrial O2⋅−from isolated mitochondria to animal models, thus offering a way to examine the many roles of mitochondrial O2⋅−production in health and disease.

中文翻译：

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MitoNeoD：针对线粒体的超氧化物探针

线粒体超氧化物（O2⋅-）是许多氧化损伤和氧化还原信号的基础。荧光探针可以检测O2-，但在体内的应用受到限制，而在细胞中，其有效性却受到副反应和DNA嵌入的限制。为了克服这些局限性，我们开发了一种双重用途的线粒体O2⋅-探针MitoNeoD，该探针可通过质谱评估体内的O2⋅-改变，并通过荧光体外评估。MitoNeoD包含一个经过修饰以防止DNA嵌入的对O2⋅-敏感的还原的菲啶鎓部分，以及一个碳-氘键以提高其对O2⋅-的选择性（通过非特异性氧化），以及一个三苯基phosph亲脂性阳离子部分，可快速积累线粒体内。