Determination of NADH in the rat brain during sleep-wake states with an optic fibre sensor and time-resolved fluorescence procedures

Mottin, Stéphane; Laporte, P.; Jouvet, Michel; Cespuglio, Raymond

doi:10.1016/s0306-4522(96)00709-9

Cited by 18 publications

(30 citation statements)

References 64 publications

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“…The fluorescence measured using our detector seems to be mostly derived from NADH, since the inclusion of a time gate and the filters used for the measurement excludes most other fluorescent substances with different fluorescence spectra, or other fluorescent substances with similar properties as NAD(P)H are present only in small concentrations (Mottin et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

“…At the end of the experiments, all animals (pharmacological experiments and CSD experiments) were killed by an overdose of the anaesthetic with the NADH measurement continuing. Animals that did not show a large and steady increase in NADH fluorescence after death (Mottin et al, 1997) were also excluded from the study.…”

Section: Procedurementioning

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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“…First attempts to determine changes in the cerebral NADH autofluorescence in nonanaesthetised animals were made throughout the sleepwake cycle, showing that the oxidative energy balance seems to be related to sleep states [26].…”

Section: Nadh Fluorescence In Freely Moving Animalsmentioning

confidence: 99%

“…However, NAD(P)H has similar optical properties as NADH and the fluorescence produced by both pyridine nucleotides can not discriminated easily. In mitochondria the concentration of NADH and the yield of the NADH derived fluorescence is higher than that of NAD(P)H [15,26,29,30]. While NADH is also distributed in the cytoplasmic fraction, it is well known that the lifetime and the yield of fluorescence of protein- bound NADH are increased significantly compared to the fluorescence of non-bound NADH [31,32].…”

Section: Introductionmentioning

confidence: 99%

Applications of laser-induced fluorescence spectroscopy for the determination of NADH in experimental neuroscience

Rex

Fink

2006

Laser Phys. Lett.

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There is an increasing need for continuously monitoring changes in brain metabolism and neuronal activity, respectively. The aim is to improve our understanding of mechanisms involved in physiological as well as pathophysiological and behavioural responses and to characterise drug actions. Changes of NADH concentration in the brain can be regarded as an index of changes in mitochondrial activity, which is closely related to neuronal activity. During the last decade the determination of NADH fluorescence by laser-induced fluorescence spectroscopy has become a method of choice in the study of mitochondrial metabolism in neuroscience. By now, small optical probes, providing excellent temporal and spatial resolution and the development of reliable and robust laser-based fluorescence detectors allow a widespread use in preclinical research. Besides in vitro studies, especially the assessment of changes in the NADH fluorescence in vivo has been shown to provide valuable information on brain function. Several applications are given, ranging from studying drug action or the extent of brain lesion to the measurement the time course of NADH concentration in a brain region of an awake and behaving laboratory rat. Theoretical aspects, opportunities, and limitations that have to be considered during the implementation of fluorescence spectroscopy are described. It is concluded, that measurement of NADH fluorescence by laserinduced fluorescence spectroscopy is a suitable tool for investigation of functional processes in the brain. Distribution and intensity of the NADH fluorescence in coronal rat brain slices containing the hippocampus. The data represent the intensity of the NADH fluorescence in arbitrary units and are shown as averaged data (n=5) from each measuring point, respectively

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“…The rapid development of the laser technique permitted the usage of lasers (nitrogen laser) instead of xenon-or mercury lamps (Renault et al, 1984). As an alternative and complementation to established methods in the neuroscience like the microdialysis technique, voltametry or potentiometric methods, optic and spectroscopic methods have been developed to assess the local metabolic state of tissue during the last years (Chance et al, 1962;Jöbsis et al, 1970;Mottin et al, 1997;Rex et al, 1999a).Nicotinamide adenine dinucleotide (NADH) is a substrate for energy transfer in the mitochondrial respiratory chain and an important parameter of the cellular metabolism (Erecinska and Wilson, 1982). Initial tests proved the reduction of spectral interference of other fluorophores by application of an early and short time gate (start +1 ns, gate with 2 ns).…”

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confidence: 99%

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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Determination of NADH in the rat brain during sleep-wake states with an optic fibre sensor and time-resolved fluorescence procedures

Cited by 18 publications

References 64 publications

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

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

Applications of laser-induced fluorescence spectroscopy for the determination of NADH in experimental neuroscience

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

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