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Age- and AD-related redox state of NADH in subcellular compartments by fluorescence lifetime imaging microscopy

  • Yue Dong
  • Michelle A. Digman
  • Gregory J. BrewerEmail author
Original Article


Nicotinamide adenine dinucleotide (reduced form: NADH) serves as a vital redox-energy currency for reduction-oxidation homeostasis and fulfilling energetic demands. While NADH exists as free and bound forms, only free NADH is utilized for complex I to power oxidative phosphorylation, especially important in neurons. Here, we studied how much free NADH remains available for energy production in mitochondria of old living neurons. We hypothesize that free NADH in neurons from old mice is lower than the levels in young mice and even lower in neurons from the 3xTg-AD Alzheimer’s disease (AD) mouse model. To assess free NADH, we used lifetime imaging of NADH autofluorescence with 2-photon excitation to be able to resolve the pool of NADH in mitochondria, cytoplasm, and nuclei. Primary neurons from old mice were characterized by a lower free/bound NADH ratio than young neurons from both non-transgenic (NTg) and more so in 3xTg-AD mice. Mitochondrial compartments maintained 26 to 41% more reducing NADH redox state than cytoplasm for each age, genotype, and sex. Aging diminished the mitochondrial free NADH concentration in NTg neurons by 43% and in 3xTg-AD by 50%. The lower free NADH with age suggests a decline in capacity to regenerate free NADH for energetic supply to power oxidative phosphorylation which further worsens in AD. Applying this non-invasive approach, we showed the most explicit measures yet of bioenergetic deficits in free NADH with aging at the subcellular level in live neurons from in-bred mice and an AD model.


NADH Aging brain Alzheimer’s disease FLIM Mitochondria Redox states 



We appreciate Prof. Enrico Gratton for inspirational discussions and the help of Rachel Cinco-Hedde, Ning Ma, and Sara Sameni at Laboratory for Fluorescence Dynamics, UC Irvine.

Funding information

This work was supported by the UC Irvine Foundation, NIH P41-GM103540 and a grant from the NIH RF1 AG058218.


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Copyright information

© American Aging Association 2019

Authors and Affiliations

  • Yue Dong
    • 1
  • Michelle A. Digman
    • 1
    • 2
  • Gregory J. Brewer
    • 1
    • 3
    Email author
  1. 1.Department of Biomedical EngineeringUniversity of California IrvineIrvineUSA
  2. 2.Laboratory of Fluorescence Dynamics, Department of Biomedical EngineeringUniversity of California IrvineIrvineUSA
  3. 3.MIND Institute, Center for Neurobiology of Learning and MemoryUniversity of CaliforniaIrvineUSA

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