期刊
CELL METABOLISM
卷 14, 期 4, 页码 555-566出版社
CELL PRESS
DOI: 10.1016/j.cmet.2011.09.004
关键词
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资金
- NSFC [31071260, 90713026, 31170815, 91013012]
- 863 Program [2006AA02Z160]
- Specialized Research Fund for the Doctoral Program of Higher Education [20100074110010]
- Fok Ying Tung Education Foundation [1110221]
- Shanghai Institutions of Higher Learning
- State Key Laboratory of Bioreactor Engineering
- 111 Project [B07023]
- NIH [HL 061795, HL 070819, HL 048743, HL 107192, HL 108630]
We have developed genetically encoded fluorescent sensors for reduced nicotinamide adenine dinucleotide (NADH), which manifest a large change in fluorescence upon NADH binding. We demonstrate the utility of these sensors in mammalian cells by monitoring the dynamic changes in NADH levels in subcellular organelles as affected by NADH transport, glucose metabolism, electron transport chain function, and redox environment, and we demonstrate the temporal separation of changes in mitochondrial and cytosolic NADH levels with perturbation. These results support the view that cytosolic NADH is sensitive to environmental changes, while mitochondria have a strong tendency to maintain physiological NADH homeostasis. These sensors provide a very good alternative to existing techniques that measure endogenous fluorescence of intracellular NAD(P)H and, owing to their superior sensitivity and specificity, allow for the selective monitoring of total cellular and compartmental responses of this essential cofactor.
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