Monitoring Cellular Metabolism with Fluorescence Lifetime of Reduced Nicotinamide Adenine Dinucleotide

Ghukasyan, Vladimir; Kao, Fu Jen

doi:10.1364/acp.2009.fe4

Cited by 21 publications

(27 citation statements)

References 5 publications

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“…Free NAD(P)H is fluorescent with an emission peak at 460 nm and a lifetime τ 1 of less than 1 nanosecond. The emission maximum of protein-bound NAD(P)H has a longer wavelength and lifetime τ 2 (2–10 nanoseconds)4950515253. If the relative contributions of the amplitudes of τ 1 and τ 2 , are a 1 for free NAD(P)H and a 2 for bound-NAD(P)H, a2/(a1 + a2) gives an estimate of the metabolic rate515253.…”

Section: Methodsmentioning

confidence: 99%

Females become infertile as the stored sperm's oxygen radicals increase

Reinhardt

Ribou

2013

Sci Rep

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Predicting infertility is central to reproductive biology, medicine and evolutionary biology. In-vitro studies suggest that oxidative sperm damage causes infertility. Oxidative sperm damage can be reduced via two fundamental pathways: the removal of oxygen radicals by antioxidants, or the interference with cell metabolism to reduce the formation of oxygen radicals. Oxidative damage protection of spermatozoa should evolve frequently, especially during female sperm storage. However, in-vivo evidence linking oxidative protection and fertility is rare. We show that the intra-sperm production rate of oxygen radicals and the sperm metabolic rate were reduced in female bedbugs, Cimex lectularius, compared to males, and females laid fertile eggs. Females became infertile when sperm oxygen radicals and sperm metabolic rate increased to male levels. Our results link female fitness to sublethal sperm damage, imply adaptive benefits of interfering with sperm metabolism and offer the hypothesis that polyandry may serve to replace low-quality sperm.

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Section: Methodsmentioning

confidence: 99%

Females become infertile as the stored sperm's oxygen radicals increase

Reinhardt

Ribou

2013

Sci Rep

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“…Between the early demonstrations in the 1990's and the present day, more than 1500 NAD(P)H fluorescence lifetime imaging studies have been published (Google Scholar), describing changes in the lifetime characteristics of live-cell NAD(P)H fluorescence in situations ranging from the onset of apoptosis [166], [170], [171], necrotic deterioration of skin [172] and wound healing [173] to stem cell differentiation [174], blood-glucose sensing [175] and aggregation of α- syn uclein in Parkinson’s disease [176]. However, until recently, the mechanisms linking the known metabolic shifts in these biological models to the changes in NAD(P)H lifetime were largely unknown, limiting their value as a biomedical assay.…”

Section: Practical Application Of Live-cell Nad(p)h Fluorescencementioning

confidence: 99%

Investigating mitochondrial redox state using NADH and NADPH autofluorescence

Blacker

Duchen

2016

Free Radical Biology and Medicine

315

276

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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.

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“…Cells and tissues contain endogenous chromophores exhibiting fluorescence called autofluorescence, and nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), and amino acids having an aromatic moiety, such as tryptophan, are known as representative autofluorescent chromophores. These chromophores exist in an extensive variety of living systems and are related to cell functions and metabolic activities [2–6]. Absorption and fluorescence spectra of representative autofluorescent chromophores are shown in Figure 1a.…”

Section: Introductionmentioning

confidence: 99%

pH Dependence of the Fluorescence Lifetime of FAD in Solution and in Cells

Islam

Honma

Nakabayashi

et al. 2013

IJMS

112

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We have studied physiological parameters in a living cell using fluorescence lifetime imaging of endogenous chromophores. In this study, pH dependence of the fluorescence lifetime of flavin adenine dinucleotide (FAD), that is a significant cofactor exhibiting autofluorescence, has been investigated in buffer solution and in cells. The fluorescence lifetime of FAD remained unchanged with pH 5 to 9 in solution. However, the fluorescence lifetime in HeLa cells was found to decrease with increasing intracellular pH, suggesting that pH in a single cell can be estimated from the fluorescence lifetime imaging of FAD without adding exogenous fluorescent probes.

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Monitoring Cellular Metabolism with Fluorescence Lifetime of Reduced Nicotinamide Adenine Dinucleotide

Cited by 21 publications

References 5 publications

Females become infertile as the stored sperm's oxygen radicals increase

Females become infertile as the stored sperm's oxygen radicals increase

Investigating mitochondrial redox state using NADH and NADPH autofluorescence

pH Dependence of the Fluorescence Lifetime of FAD in Solution and in Cells

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