Gilles Bloch

Early N-Acetylaspartate Depletion Is a Marker of Neuronal Dysfunction in Rats and Primates Chronically Treated with the Mitochondrial Toxin 3-Nitropropionic Acid:

N-acetylaspartate (NAA) quantification by 1H-magnetic resonance spectroscopy has been commonly used to assess in vivo neuronal loss in neurodegenerative disorders. Here, the authors used ex vivo and in vivo1H-magnetic resonance spectroscopy in rat and primate models of progressive striatal degeneration induced by the mitochondrial toxin 3-nitropropionate (3NP) to determine whether early NAA depletions could also be associated with neuronal dysfunction. In rats that were treated for 3 days with 3NP and had motor symptoms, the authors found a significant decrease in NAA concentrations, specifically restricted to the striatum. No cell loss or dying cells were found at this stage in these animals. After 5 days of 3NP treatment, a further decrease in striatal NAA concentrations was observed in association with the occurrence of dying neurons in the dorsolateral striatum. In 3NP-treated primates, a similar striatal-selective and early decrease in NAA concentrations was observed after only a few weeks of neurotoxic treatment, without any sign of ongoing cell death. This early decrease in striatal NAA was partially reversed after 4 weeks of 3NP withdrawal. These results demonstrate that early NAA depletions reflect a reversible state of neuronal dysfunction preceding cell degeneration and suggest that in vivo quantification of NAA 1H-magnetic resonance spectroscopy may become a valuable tool for assessing early neuronal dysfunction and the effects of potential neuroprotective therapies in neurodegenerative disorders.

Brain gaba editing without macromolecule contamination

A new scheme is proposed to edit the 3.0 ppm GABA resonance without macromolecule (MM) contamination. Like previous difference spectroscopy approaches, the new scheme manipulates J‐modulation of this signal using a selective editing pulse. The elimination of undesirable MM contribution at 3.0 ppm is obtained by applying this pulse symmetrically about the J‐coupled MM resonance, at 1.7 ppm, in the two steps of the editing scheme. The effectiveness of the method is demonstrated in vitro, using lysine to mimic MM, and in vivo. As compared to the most commonly used editing scheme, which necessitates the acquisition and processing of two distinct difference spectroscopy experiments, the new scheme offers a reduction in experimental time (–33%) and an increase in accuracy. Magn Reson Med 45:517–520, 2001.

Decreased TCA cycle rate in the rat brain after acute 3-NP treatment measured by in vivo 1H-[13C] NMR spectroscopy.

Inhibition of succinate dehydrogenase (SDH) by the mitochondrial toxin 3‐nitropropionic acid (3‐NP) has gained acceptance as an animal model of Huntingtons disease. In this study 13C NMR spectroscopy was used to measure the tricarboxylic acid (TCA) cycle rate in the rat brain after 3‐NP treatment. The time course of both glutamate C4 and C3 13C labelling was monitored in vivo during an infusion of [1‐13C]glucose. Data were fitted by a mathematical model to yield the TCA cycle rate (Vtca) and the exchange rate between α‐ketoglutarate and glutamate (Vx). 3‐NP treatment induced a 18% decrease in Vtca from 0.71 ± 0.02 µmol/g/min in the control group to 0.58 ± 0.02 µmol/g/min in the 3‐NP group (p < 0.001). Vx increased from 0.88 ± 0.08 µmol/g/min in the control group to 1.33 ± 0.24 µmol/g/min in the 3‐NP group (p < 0.07). Fitting the C4 glutamate time course alone under the assumption that Vx is much higher than Vtca yielded Vtca=0.43 µmol/g/min in both groups. These results suggest that both Vtca and Vx are altered during 3‐NP treatment, and that both glutamate C4 and C3 labelling time courses are necessary to obtain a reliable measurement of Vtca.

Field-Frequency Locked In Vivo Proton MRS on a Whole-Body Spectrometer

The stability of the main magnetic field is critical for prolonged in vivo magnetic resonance spectroscopy (MRS) acquisitions, especially for difference spectroscopy. This study was focused on the implementation and optimization of a field‐frequency lock (FFL) on a whole body spectrometer, to correct the main field drift during localized proton MRS of the human brain. The FFL was achieved through a negative feed‐back applied in real time on the Z0 shim coil current, after calculation of the frequency shift from a reference signal. This signal was obtained from the whole head with a small flip angle acquisition interleaved with the PRESS acquisition of interest. To avoid propagation of the important short‐term time‐correlated fluctuations of the head water frequency (mainly due to respiratory motion) onto Z0 correction, the sampling rate of the reference frequency and the smoothing window for the Z0 correction were carefully optimized. Thus, an effective FFL was demonstrated in vivo with no significant increase of the short‐term variance of the water frequency. Magn Reson Med 1999 42:636–642, 1999.

In vivo quantification of localized neuronal activation and inhibition in the rat brain using a dedicated high temporal-resolution β+-sensitive microprobe

Understanding brain disorders, the neural processes implicated in cognitive functions and their alterations in neurodegenerative pathologies, or testing new therapies for these diseases would benefit greatly from combined use of an increasing number of rodent models and neuroimaging methods specifically adapted to the rodent brain. Besides magnetic resonance (MR) imaging and functional MR, positron-emission tomography (PET) remains a unique methodology to study in vivo brain processes. However, current high spatial-resolution tomographs suffer from several technical limitations such as high cost, low sensitivity, and the need of restraining the animal during image acquisition. We have developed a β+-sensitive high temporal-resolution system that overcomes these problems and allows the in vivo quantification of cerebral biochemical processes in rodents. This β-MICROPROBE is an in situ technique involving the insertion of a fine probe into brain tissue in a way very similar to that used for microdialysis and cell electrode recordings. In this respect, it provides information on molecular interactions and pathways, which is complementary to that produced by these technologies as well as other modalities such as MR or fluorescence imaging. This study describes two experiments that provide a proof of concept to substantiate the potential of this technique and demonstrate the feasibility of quantifying brain activation or metabolic depression in individual living rats with 2-[18F]fluoro-2-deoxy-d-glucose and standard compartmental modeling techniques. Furthermore, it was possible to identify correctly the origin of variations in glucose consumption at the hexokinase level, which demonstrate the strength of the method and its adequacy for in vivo quantitative metabolic studies in small animals.

Multimodal neuroimaging provides a highly consistent picture of energy metabolism, validating 31P MRS for measuring brain ATP synthesis

Neuroimaging methods have considerably developed over the last decades and offer various noninvasive approaches for measuring cerebral metabolic fluxes connected to energy metabolism, including PET and magnetic resonance spectroscopy (MRS). Among these methods, 31P MRS has the particularity and advantage to directly measure cerebral ATP synthesis without injection of labeled precursor. However, this approach is methodologically challenging, and further validation studies are required to establish 31P MRS as a robust method to measure brain energy synthesis. In the present study, we performed a multimodal imaging study based on the combination of 3 neuroimaging techniques, which allowed us to obtain an integrated picture of brain energy metabolism and, at the same time, to validate the saturation transfer 31P MRS method as a quantitative measurement of brain ATP synthesis. A total of 29 imaging sessions were conducted to measure glucose consumption (CMRglc), TCA cycle flux (VTCA), and the rate of ATP synthesis (VATP) in primate monkeys by using 18F-FDG PET scan, indirect 13C MRS, and saturation transfer 31P MRS, respectively. These 3 complementary measurements were performed within the exact same area of the brain under identical physiological conditions, leading to: CMRglc = 0.27 ± 0.07 μmol·g−1·min−1, VTCA = 0.63 ± 0.12 μmol·g−1·min−1, and VATP = 7.8 ± 2.3 μmol·g−1·min−1. The consistency of these 3 fluxes with literature and, more interestingly, one with each other, demonstrates the robustness of saturation transfer 31P MRS for directly evaluating ATP synthesis in the living brain.

NMR measurement of brain oxidative metabolism in monkeys using 13C-labeled glucose without a 13C radiofrequency channel

We detected glutamate C4 and C3 labeling in the monkey brain during an infusion of [U‐13C6]glucose, using a simple 1H PRESS sequence without 13C editing or decoupling. Point‐resolved spectroscopy (PRESS) spectra revealed decreases in 12C‐bonded protons, and increases in 13C‐bonded protons of glutamate. To take full advantage of the simultaneous detection of 12C‐ and 13C‐bonded protons, we implemented a quantitation procedure to properly measure both glutamate C4 and C3 enrichments. This procedure relies on LCModel analysis with a basis set to account for simultaneous signal changes of protons bound to 12C and 13C. Signal changes were mainly attributed to 12C‐ and 13C‐bonded protons of glutamate. As a result, we were able to measure the tricarboxylic acid (TCA) cycle flux in a 3.9 cm3 voxel centered in the monkey brain on a whole‐body 3 Tesla system (VTCA = 0.55 ± 0.04 μmol.g−1.min−1, N = 4). This work demonstrates that oxidative metabolism can be quantified in deep structures of the brain on clinical MRI systems, without the need for a 13C radiofrequency (RF) channel. Magn Reson Med 52:33–40, 2004.

How to investigate oxygen supply, uptake, and utilization simultaneously by interleaved NMR imaging and spectroscopy of the skeletal muscle

Human skeletal muscle perfusion, oxygenation, and high‐energy phosphate distribution were measured simultaneously by interleaved 1H and 31P NMR spectroscopy and 1H NMR imaging in vivo. From these parameters, arterial oxygen supply (DO2), muscle reoxygenation rate, mitochondrial ATP production, and O2 consumption (VO2) were deduced at the recovery phase of a short ischemic exercise bout. In addition, by using a reformulation of the mass conservation law, muscle maximum O2 extraction was calculated from these parameters. Magn Reson Med, 2005.

pH as a Biomarker of Neurodegeneration in Huntington's Disease: A Translational Rodent-Human MRS Study

Early diagnosis and follow-up of neurodegenerative diseases are often hampered by the lack of reliable biomarkers. Neuroimaging techniques like magnetic resonance spectroscopy (MRS) offer promising tools to detect biochemical alterations at early stages of degeneration. Intracellular pH, which can be measured noninvasively by 31P-MRS, has shown variations in several brain diseases. Our purpose has been to evaluate the potential of MRS-measured pH as a relevant biomarker of early degeneration in Huntingtons disease (HD). We used a translational approach starting with a preclinical validation of our hypothesis before adapting the method to HD patients. 31P-MRS-derived cerebral pH was first measured in rodents during chronic intoxication with 3-nitropropionic acid (3NP). A significant pH increase was observed early into the intoxication protocol (pH = 7.17 ± 0.02 after 3 days) as compared with preintoxication (pH = 7.08 ± 0.03). Furthermore, pH changes correlated with the 3NP-induced inhibition of succinate dehydrogenase and preceded striatum lesions. Using a similar MRS approach implemented on a clinical MRI, we then showed that cerebral pH was significantly higher in HD patients (n = 7) than in healthy controls (n = 6) (7.05 ± 0.03 versus 7.02 ± 0.01, respectively, P = 0.026). Altogether, both preclinical and human data strongly argue in favor of MRS-measured pH being a promising biomarker for diagnosis and follow-up of HD.

Diffusion-weighted NMR spectroscopy allows probing of 13C labeling of glutamate inside distinct metabolic compartments in the brain.

In the present work, diffusion‐weighted (DW)‐NMR spectroscopy of glutamate was performed during a 13C‐labeled glucose infusion in monkey brain (six experiments). It is shown that glutamate 13C labeling occurs significantly faster at higher diffusion weightings—slightly for glutamate in position C4, and more markedly for glutamate in position C3. This demonstrates the existence of different diffusion compartments for glutamate, associated with different metabolic rates. Metabolic modeling of 13C enrichment time‐courses suggests that these compartments might be gray and white matter, each having a specific oxidative metabolism rate possibly paralleled by a specific glutamate diffusion coefficient. Magn Reson Med 60:306–311, 2008.