Cardiac metabolism monitored by fiberoptic laser fluorimeter

Renault, G.; Raynal, E.; Sinet, Martine; Muffat-Joly, Martine; Vallois, J M; Berthier, J. P.; Cornillault, J.; Godard, Bruno

doi:10.1016/0002-8703(84)90644-6

Cited by 8 publications

(5 citation statements)

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“…The strength of the NADH fluorescence signal is proportional to the concentration of the NADH molecules. Various fluorometric methods have been developed with high powered light sources such as xenon lamps, tungsten‐halogen lamps, mercury arc lamps or lasers (Renault et al 1984; Ince et al 1992). In this study a nitrogen‐laser‐fluorescence detector with spectral filters has been used to induce and measure NADH fluorescence in the cortex of rats.…”

mentioning

confidence: 99%

Bioavailability of Reduced Nicotinamide‐adenin‐dinucleotide (NADH) in the Central Nervous System of the Anaesthetized Rat Measured by Laser‐Induced Fluorescence Spectroscopy

Rex

Hentschke

Fink

2002

Pharmacology & Toxicology

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Drugs intended to increase wellness or quality of life (''lifestyle drugs'') have gained popularity and/or importance over recent years. Biogenic substances like nicotinamide adenine dinucleotide (NADH) are supposed to increase the physical and intellectual performance without side-effects. NADH is an energy-delivering co-substrate in the respiratory chain. Clinical studies showed positive effects of peripherally given NADH in Morbus Parkinson and major depression. NADH can be measured by its fluorescence. In this study a pulsed N 2 -laser combined with a fibre-optic probe and photomultipliers was used to induce and measure NADH fluorescence in the rat cortex. The aims of the study were to assess the suitability of the laser-induced spectroscopy for in vivo and on-line measurement of NADH in neuroscience and the assessment of the central availability of NADH after peripheral administration. NADH (50 mg/kg) but not the precursor nicotinamide caused a significant rise of the NADH fluorescence intensity indicating an increase of the NADH concentration in the rat cortex. In conclusion, the results suggest that NADH given orally or intraperitoneally increases the amounts of NADH in the brain. The results may thus help to explain the clinical effects reported.Nicotinamide adenine dinucleotide (NADH) plays a central role as a co-substrate for energy transfer in the mitochondrial oxidative phosphorylation, which supplies more than 95% of the total energy requirement in the cells (Erecinska & Wilson 1982). In most cells NADH is the main source of reducing equivalents and NADH is therefore an important factor of cellular metabolism. The vitamin B3 nicotinamide is the main precursor for the synthesis of NADH molecules in eucariotic cells.There is evidence that an additionally supply of co-factors or substrates could have beneficial effects in the diseases of the central nervous system. NADH as an energysupplying substrate could also support enzymatic reactions. Clinical studies demonstrate the positive effect of peripherally administered NADH on serious diseases such as Parkinson's disease, chronic fatigue syndrom and major depression (Birkmayer & Birkmayer 1991;Birkmayer et al. 1993;Forsyth et al. 1999).In connection with these clinical studies which show central effects after peripheral administration, we wanted to investigate whether peripherally administered NADH might change the central concentration of NADH.Selective determination of NADH in vivo is possible due to its spectroscopic properties. NADH and the corresponding oxidized molecule NADπ are natural fluorophores, which start to fluoresce after stimulation. The reduced and energy-rich form NADH has two absorption peaks at (lΩ 260 nm and 340 nm), the oxidized form has only one absorption peak at (lΩ260 nm). Stimulation of the molecules with light at wavelengths around 340 nm (ultraviolet-spectrum) leads to generation of fluorescence signals (maximum at 465 nm) that start only from the reduced molecules. The strength of the NADH fluorescence signal is pr...

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confidence: 99%

Bioavailability of Reduced Nicotinamide‐adenin‐dinucleotide (NADH) in the Central Nervous System of the Anaesthetized Rat Measured by Laser‐Induced Fluorescence Spectroscopy

Rex

Hentschke

Fink

2002

Pharmacology & Toxicology

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“…Methods for the determination of NADH fluorescence are well established (e. g. Chance et al, 1962;Renault et al, 1984, Ince et al, 1992. Using a small optical probe with a minor penetration depth of the excitation light allows a high spatial resolution and is effective in reducing the background (Beuthan et al, 1990;Fink et al, 1993).…”

Section: Fig 2 Time-resolved Laser Fluorescence Spectra Of Nadhmentioning

confidence: 99%

“…NAD + /NADH has a spectroscopically interesting property: the reduced form NADH absorbs light at wavelengths of λ=260 and λ=340 nm, whereas the oxidised form NAD + has only one absorption maximum at λ=260 nm. The rapid development of the laser technique permitted the usage of lasers (nitrogen laser) instead of xenon-or mercury lamps (Renault et al, 1984). Measurement of the signal intensity enables the estimation of changes in the NAD + /NADH ratio (Ince et al, 1992).…”

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confidence: 99%

“…Later, flexible fibre-optic light guides have been applied, eliminating the limitations of a rigid optical set-up (Mayewsky, 1984). The rapid development of the laser technique permitted the usage of lasers (nitrogen laser) instead of xenon-or mercury lamps (Renault et al, 1984). Recently, the development of small and efficient lasers enabled the construction of compact and transportable fluorescence detection systems designed for medical research and diagnostic purposes (Fink et al, 1993).…”

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Determination of NADH in Frozen Rat Brain Sections by Laser-Induced Fluorescence

Rex¹,

Fink²

2001

Biological Chemistry

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Methods to assess metabolism are important analytical tools in neuroscience. The fluorophore nicotinamide adenine dinucleotide (NADH) is a parameter of cellular metabolism. NADH fluorescence was measured using a laser-based fluorescence detector with spectral and temporal filters. Distribution and intensity of NADH fluorescence were investigated in frozen brain sections. In sections containing hippocampus the intensity of NADH fluorescence was correlated to brain structures. In order to investigate the consequences of neurotoxic lesions, 5,7-dihydroxytryptamine was injected into the dorsal raphe nucleus 4 to 240 days prior to the measurement. NADH fluorescence decreased in the affected region by 50%, indicating that no recovery in metabolic activity had occurred.

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“…In this study, a laser-based fluorescence detector (Renault et al, 1984;Beuthan et al, 1990;Fink et al, 1993) was used to induce and measure NADH fluorescence on the cortical surface. The application of a short pulsed nitrogen laser as excitation source allows usage of single optical fibres with a diameter of 100 µm in combination with a time-gated signal recognition.…”

Section: Introductionmentioning

confidence: 99%

Cortical NADH during pharmacological manipulations of the respiratory chain and spreading depression in vivo

Rex

Fink

1999

J. Neurosci. Res.

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The nicotinamide adenine dinucleotide (NADH) is one of the main means for energy transfer in the mitochondrial respiratory chain and is an important parameter of cellular metabolism. NADH can be measured by its fluorescence and various fluorometric methods have been developed. In this study, a pulsed nitrogen laser combined with a fibreoptic set-up and photomultipliers was used to induce and measure NADH fluorescence on the cortical surface. The aim of the study was to assess the suitability of the laser induced spectroscopy for in vivo and on-line measurement of NADH in neuroscience and particularly for the assessment of neuronal metabolism. Changes in cerebral blood flow may affect fluorescence measurement. To assess the consequences of alterations in blood flow, the vasodilators glyceryl trinitrate and nimodipine and the vasoconstrictor endothelin-1 were applied. The induced hemodynamic changes were verified by colour Doppler sonography. The tests using the vasodilators showed that an increased blood flow in the brain increased not only NADH fluorescence but also the scattered light measured. The vasoconstrictor caused opposite effects. Insertion of a compensation method (subtraction of the scattered light) allowed the exclusion of hemodynamic artifacts. Effects of changes in the cellular metabolism were induced by sodium cyanide, an inhibitor of the mitochondrial respiratory chain, or by 2,4-dinitrophenol (2,4-DNP), an uncoupler of the oxidative phosphorylation. Sodium cyanide induced a transient increase of NADH fluorescence and 2,4-DNP decreased intracellular NADH fluorescence. Furthermore, the repercussions of cortical spreading depressions (CSD), a response of the brain to noxious stimuli, on cortical NADH fluorescence were determined. A single CSD decreased cortical NADH fluorescence for about 1 min, followed by a 5- to 10-min increase. The changes in NADH levels seem to correspond with the excitation and inhibition of neuronal metabolism, respectively. In summary, the measurement of NADH fluorescence using the laser technique allows the determination of changes in oxidative phosphorylation with high regional selectivity and time resolution.

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Cardiac metabolism monitored by fiberoptic laser fluorimeter

Cited by 8 publications

References 1 publication

Bioavailability of Reduced Nicotinamide‐adenin‐dinucleotide (NADH) in the Central Nervous System of the Anaesthetized Rat Measured by Laser‐Induced Fluorescence Spectroscopy

Bioavailability of Reduced Nicotinamide‐adenin‐dinucleotide (NADH) in the Central Nervous System of the Anaesthetized Rat Measured by Laser‐Induced Fluorescence Spectroscopy

Determination of NADH in Frozen Rat Brain Sections by Laser-Induced Fluorescence

Cortical NADH during pharmacological manipulations of the respiratory chain and spreading depression in vivo

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