Coupling of neuronal activity and mitochondrial metabolism as revealed by nad(p)h fluorescence signals in organotypic hippocampal slice cultures of the rat

Kann, Oliver; Schuchmann, Sebastian; Buchheim, Katharina; Heinemann, Uwe

doi:10.1016/s0306-4522(03)00026-5

Cited by 92 publications

(124 citation statements)

References 58 publications

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“…From Figure 8A, it appears that the decrease in overshoot amplitude may be explicable in part because a ceiling level had been reached. Selective decreases in the amplitude of the overshoot component of NAD(P)H transients have been described previously, when repetitive stimuli are applied at the peak of overshoot responses in isolated neurons (Duchen, 1992) and in the hippocampal slice preparation (Kann et al, 2003).…”

Section: Effects Of Pyruvatesupporting

confidence: 61%

NAD(P)H Fluorescence Transients after Synaptic Activity in Brain Slices: Predominant Role of Mitochondrial Function

Brennan

Connor

Shuttleworth

2006

J Cereb Blood Flow Metab

115

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Excitatory stimulation in hippocampal slices results in biphasic NAD(P)H fluorescence transients. Previous studies using differing stimulus protocols agreed that the oxidation phase is a consequence of mitochondrial metabolism, but the reduction phase has been attributed to (1) mitochondrial nicotinamide adenine dinucleotide (NADH) generation or (2) astrocytic glycolysis triggered by glutamate uptake. In an attempt to reconcile these two views, the present study examined NAD(P)H signals evoked by a wide range of stimulus durations (40 ms to 20 secs). A combination of ionotropic glutamate receptor (iGluR) antagonists (6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-amino-5-phosphonopentanoic acid (APV)) virtually abolished responses to brief stimuli (40 to 200 ms, 50 Hz), but a significant fraction of the signal elicited by extended stimulation (20 secs, 32 Hz) was resistant to CNQX/APV. Glycolysis was inhibited by removal of glucose and addition of 2-deoxyglucose (2DG) (10 mmol/L) or iodoacetic acid (IAA, 1 mmol/L). Pyruvate was provided as an alternative substrate for oxidative phosphorylation and the A1 receptor antagonist 1,3-Dipropyl-8-cyclopentylxanthine (DPCPX) included to prevent decreases in synaptic efficacy. If sufficient pyruvate was supplied, responses to brief and extended stimuli were unaffected by glycolytic inhibition and not significantly reduced by an inhibitor of glucose uptake (3-O-methyl glucose, 3 mmol/L). When timed to arrive at the peak of overshoots generated by extended synaptic stimulation, brief pyruvate applications (10 mmol/L, 2 mins) had little effect on evoked NAD(P)H increases. Flavoprotein autofluorescence transients after extended stimuli matched (with inverted sign) NAD(P)H responses. Responses to extended stimuli were not reduced by a nonselective inhibitor of glutamate uptake DL-Threo-b-benzyloxyaspartic acid (TBOA). These results suggest that NAD(P)H transients report mitochondrial dynamics, rather than recruitment of glycolytic metabolism, over a wide range of stimulus intensities.

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Section: Effects Of Pyruvatesupporting

confidence: 61%

NAD(P)H Fluorescence Transients after Synaptic Activity in Brain Slices: Predominant Role of Mitochondrial Function

Brennan

Connor

Shuttleworth

2006

J Cereb Blood Flow Metab

115

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“…Rhod2 is known to load into the cytosolic (del Nido et al, 1998;MacGowan et al, 2001a;Rubart et al, 2003) as well as the mitochondrial compartment (Hajnoczky et al, 1995;Kann et al, 2003;Scaduto and Grotyohann, 2003;Mironov et al, 2004), a feature which might complicate quantification and interpretation of Rhod2-Ca 2 þ fluorescence changes. Minta et al (1989) originally described that Rhod2 accumulate, hydrolyzed and trapped in the mitochondrial compartment.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

“…Although cell culture experiments may differ fundamentally from those operating in vivo these results are important for the interpretation of our findings obtained during and after ischemia. Recently, Kann et al (2003) concluded that most (79%) of the Rhod2 signal came from the mitochondrial compartments. They used a low dose of Rhod2/AM (5 mmol/L) and loaded it into brain slice culture at 36.5 1C for B50 to 60 mins.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

“…In excitable cells, calcium fluxes are closely linked to electrical activity (Helmchen and Waters, 2002). Thus, measurements of intracellular calcium directly track neuronal electrical activity and in vitro studies on single cells (Duchen, 1992;Miyata et al, 1994;Takahashi et al, 1993) and brain slice preparations (Kudo et al, 1992;Kann et al, 2003) have confirmed this. Two-photon microscopy has also been used to monitor calcium changes in superficial brain regions in vivo (Helmchen and Waters, 2002).…”

Section: Introductionmentioning

confidence: 92%

“…Other investigators have used Rhod2 in studies of single neurons, brain slices and the brain in vivo (Takahashi et al, 1999;Kudo, 1996;Takita et al, 2004). It is clear from these studies that Rhod2 loads into the cytosolic (del Nido et al, 1998;MacGowan et al, 2001a) as well as mitochondrial compartment (Kann et al, 2003;Scaduto and Grotyohann, 2003), a feature which might complicate quantification and interpretation of Rhod2-Ca 2 þ -sensitive fluorescence changes.…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous Detection of Blood Volume, Oxygenation, and Intracellular Calcium Changes during Cerebral Ischemia and Reperfusionin vivoUsing Diffuse Reflectance and Fluorescence

Koretsky

Izrailtyan

et al. 2005

J Cereb Blood Flow Metab

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We describe an approach to measure changes in intracellular calcium along with changes in blood volume and oxygenation directly from the exposed rat cortex in vivo during cerebral ischemia and reperfusion. Measurements were made using a catheter-based optical system. The endface of a Yshaped bifurcated fiber optic bundle was mounted on the cortical surface. It delivered the light at three wavelengths of 548, 555, and 572 nm to the brain through a fast monochromator coupled to a xenon lamp, and collected the calcium-dependent fluorescence emission from Rhod2 at 589 nm (excited at 548 nm) along with the diffuse reflections at the wavelengths of 555 and 572 nm to determine the changes in blood volume and hemoglobin oxygenation. The feasibility of this approach was experimentally examined by inducing transient cerebral ischemia and reperfusion in the rat. The ischemia induced an 8.5%71.7% fluorescence increase compared with the preischemic control values. Blood volume and tissue hemoglobin oxygenation decreased by 57.4%712.6% and 47.3%712.5%, respectively. All signals normalized on reperfusion. The ischemia-induced change in Rhod2-Ca 2 þ fluorescence was blocked using a calcium channel blocker, nimodipine, confirming that intracellular changes in calcium were responsible for the fluorescence changes. Thus, changes in cerebral hemodynamics and intracellular calcium concentration changes were measured simultaneously, facilitating future studies of the interrelationship between neuronal activation and metabolic and vascular processes in normal and diseased brain.

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NBCe1 mediates the regulation of the NADH/NAD⁺redox state in cortical astrocytes by neuronal signals

Köhler

Winkler

Sicker

et al. 2018

Glia

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Astrocytes are a glial cell type, which is indispensable for brain energy metabolism. Within cells, the NADH/NAD redox state is a crucial node in metabolism connecting catabolic pathways to oxidative phosphorylation and ATP production in mitochondria. To characterize the dynamics of the intracellular NADH/NAD redox state in cortical astrocytes Peredox, a genetically encoded sensor for the NADH/NAD redox state, was expressed in cultured cortical astrocytes as well as in cortical astrocytes in acutely isolated brain slices. Calibration of the sensor in cultured astrocytes revealed a mean basal cytosolic NADH/NAD redox ratio of about 0.01; however, with a broad distribution and heterogeneity in the cell population, which was mirrored by a heterogeneous basal cellular concentration of lactate. Inhibition of glucose uptake decreased the NADH/NAD redox state while inhibition of lactate dehydrogenase or of lactate release resulted in an increase in the NADH/NAD redox ratio. Furthermore, the NADH/NAD redox state was regulated by the extracellular concentration of K , and application of the neurotransmitters ATP or glutamate increased the NADH/NAD redox state dependent on purinergic receptors and glutamate uptake, respectively. This regulation by K , ATP, and glutamate involved NBCe1 mediated sodium-bicarbonate transport. These results demonstrate that the NADH/NAD redox state in astrocytes is a metabolic node regulated by neuronal signals reflecting physiological activity, most likely contributing to adjust astrocytic metabolism to energy demand of the brain.

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Coupling of neuronal activity and mitochondrial metabolism as revealed by nad(p)h fluorescence signals in organotypic hippocampal slice cultures of the rat

Cited by 92 publications

References 58 publications

NAD(P)H Fluorescence Transients after Synaptic Activity in Brain Slices: Predominant Role of Mitochondrial Function

NAD(P)H Fluorescence Transients after Synaptic Activity in Brain Slices: Predominant Role of Mitochondrial Function

Simultaneous Detection of Blood Volume, Oxygenation, and Intracellular Calcium Changes during Cerebral Ischemia and Reperfusionin vivoUsing Diffuse Reflectance and Fluorescence

NBCe1 mediates the regulation of the NADH/NAD⁺redox state in cortical astrocytes by neuronal signals

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Coupling of neuronal activity and mitochondrial metabolism as revealed by nad(p)h fluorescence signals in organotypic hippocampal slice cultures of the rat

Cited by 92 publications

References 58 publications

NAD(P)H Fluorescence Transients after Synaptic Activity in Brain Slices: Predominant Role of Mitochondrial Function

NAD(P)H Fluorescence Transients after Synaptic Activity in Brain Slices: Predominant Role of Mitochondrial Function

Simultaneous Detection of Blood Volume, Oxygenation, and Intracellular Calcium Changes during Cerebral Ischemia and Reperfusionin vivoUsing Diffuse Reflectance and Fluorescence

NBCe1 mediates the regulation of the NADH/NAD+redox state in cortical astrocytes by neuronal signals

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NBCe1 mediates the regulation of the NADH/NAD⁺redox state in cortical astrocytes by neuronal signals