Investigating mitochondrial redox state using NADH and NADPH autofluorescence.
Investigating mitochondrial redox state using NADH and NADPH autofluorescence.
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DOI：
10.1016/j.freeradbiomed.2016.08.010
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发表时间：
2016-11
影响因子：
7.4
通讯作者：
Duchen, Michael R.
中科院分区：
文献类型：
--
作者：
Blacker, Thomas S.;Duchen, Michael R.
NAD和NADP吡啶核苷酸库的氧化还原状态在确定能量产生途径的活性、驱动氧化应激和维持抗氧化防御方面起着关键作用。一般来说，NAD主要参与调节产生能量的分解代谢过程，而NADP可能参与抗氧化防御和自由基的产生。这些通路平衡的缺陷与许多疾病有关，从糖尿病、神经退行性疾病到心脏病和癌症。因此，评估活组织中这些单独池的丰度和氧化还原状态的方法将为潜在的病理生理学提供宝贵的见解。实验上，自二十世纪中期以来，氧化还原辅助因子NADH和NADPH的还原形式的固有荧光已被用于此目的。在这篇综述中，我们概述了这些技术在复杂组织制剂中研究线粒体氧化还原状态的现代实现。由于NADH和NADPH的荧光光谱难以区分，因此解释它们联合荧光（通常标记为NAD(P)H）产生的信号可能很复杂。因此，我们讨论了最近使用荧光寿命成像显微镜（FLIM）的研究，它提供了区分两个不同池的潜力。该技术在生物医学研究中提供了细胞自身荧光增加的代谢信息，为在病变组织中观察到的时间分辨NAD(P)H荧光信号的变化提供了生化见解。NAD在能量产生途径中起着核心作用。NADP对维持抗氧化防御至关重要。还原形式，NADH和NADPH，在活组织内自然荧光。细胞自身荧光可用于研究NAD(P)H氧化还原状态。NAD(P)H FLIM允许NADH和NADPH的贡献分离。
The redox states of the NAD and NADP pyridine nucleotide pools play critical roles in defining the activity of energy producing pathways, in driving oxidative stress and in maintaining antioxidant defences. Broadly speaking, NAD is primarily engaged in regulating energy-producing catabolic processes, whilst NADP may be involved in both antioxidant defence and free radical generation. Defects in the balance of these pathways are associated with numerous diseases, from diabetes and neurodegenerative disease to heart disease and cancer. As such, a method to assess the abundance and redox state of these separate pools in living tissues would provide invaluable insight into the underlying pathophysiology. Experimentally, the intrinsic fluorescence of the reduced forms of both redox cofactors, NADH and NADPH, has been used for this purpose since the mid-twentieth century. In this review, we outline the modern implementation of these techniques for studying mitochondrial redox state in complex tissue preparations. As the fluorescence spectra of NADH and NADPH are indistinguishable, interpreting the signals resulting from their combined fluorescence, often labelled NAD(P)H, can be complex. We therefore discuss recent studies using fluorescence lifetime imaging microscopy (FLIM) which offer the potential to discriminate between the two separate pools. This technique provides increased metabolic information from cellular autofluorescence in biomedical investigations, offering biochemical insights into the changes in time-resolved NAD(P)H fluorescence signals observed in diseased tissues. NAD plays a central role in energy-producing pathways. NADP is crucial for maintaining the antioxidant defence. The reduced forms, NADH and NADPH, are naturally fluorescent inside living tissues. Cellular autofluorescence can be used to investigate NAD(P)H redox state. NAD(P)H FLIM allows the contributions from NADH and NADPH to be separated.
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DOI：
10.1073/pnas.0708959104
发表时间：
2007-11-13
影响因子：
11.1
作者：
Abramov, Andrey Y.;Fraley, Cresson;Pavlov, Evgeny
通讯作者：
Pavlov, Evgeny
影响因子：
6.3
作者：
Berndt, Nikolaus;Kann, Oliver;Holzhuetter, Hermann-Georg
通讯作者：
Holzhuetter, Hermann-Georg
影响因子：
3.7
作者：
Caldwell, Matthew;Moroz, Tracy;Tachtsidis, Ilias
通讯作者：
Tachtsidis, Ilias
影响因子：
64.8
作者：
Chouchani, Edward T.;Pell, Victoria R.;Gaude, Edoardo;Aksentijevic, Dunja;Sundier, Stephanie Y.;Robb, Ellen L.;Logan, Angela;Nadtochiy, Sergiy M.;Ord, Emily N. J.;Smith, Anthony C.;Eyassu, Filmon;Shirley, Rachel;Hu, Chou-Hui;Dare, Anna J.;James, Andrew M.;Rogatti, Sebastian;Hartley, Richard C.;Eaton, Simon;Costa, Ana S. H.;Brookes, Paul S.;Davidson, Sean M.;Duchen, Michael R.;Saeb-Parsy, Kourosh;Shattock, Michael J.;Robinson, Alan J.;Work, Lorraine M.;Frezza, Christian;Krieg, Thomas;Murphy, Michael P.
通讯作者：
Murphy, Michael P.
影响因子：
29
作者：
Alves TC;Pongratz RL;Zhao X;Yarborough O;Sereda S;Shirihai O;Cline GW;Mason G;Kibbey RG
通讯作者：
Kibbey RG