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Label Incorporation (label + incorporation)
Selected AbstractsAstrocyte metabolism is disturbed in the early development of experimental hydrocephalusJOURNAL OF NEUROCHEMISTRY, Issue 1 2003Daniel Kondziella Abstract The proper diagnosis of the arrested or the progressive form of hydrocephalus has a critical impact on treatment, but remains difficult. The assessment of early changes in cerebral metabolism might help in the development of adequate non-invasive diagnostic tools. This study examined the alterations in label incorporation in neurotransmitter amino acids and other compounds in kaolin-induced progressive hydrocephalus in rats by means of magnetic resonance spectroscopy (MRS) combined with the administration of [1- 13C]glucose and [1,2- 13C]acetate. Some 2, 4 and 6 weeks after kaolin injection into the cisterna magna, cerebrum, brainstem and cerebellum were dissected. Interestingly, labelling of most amino acids derived from [1- 13C]glucose showed no alterations, whereas labelling from [1,2- 13C]acetate was affected. Two weeks after induction of hydrocephalus the taurine concentration was decreased, whereas the concentration of [1,2- 13C]lactate was increased in the cerebrum and that of [1,2- 13C]GABA in the brainstem. Furthermore, labelling from [1,2- 13C]acetate was significantly decreased in [4,5- 13C]glutamate, [1,2- 13C]glutamate and [1,2- 13C]GABA in cerebrum from 4 weeks after hydrocephalus induction. The concentration of N -acetylaspartate, a neuronal marker, was unchanged. However, labelling of the acetyl group from [1- 13C]glucose was decreased in cerebellum and brainstem at 6 weeks after the induction of hydrocephalus. As glucose is metabolized predominately by neurones, whereas acetate is exclusively taken up by astrocytes, these results indicate that mostly astrocytic, and only later neuronal, metabolism is disturbed in the kaolin model of hydrocephalus. If verified in patients using in vivo MRS, impaired astrocyte metabolism might serve as an early indication for operative treatment. [source] Cultured Granule Cells and Astrocytes from Cerebellum Differ in Metabolizing SphingosineJOURNAL OF NEUROCHEMISTRY, Issue 2 2000Laura Riboni Sphingosine metabolism was studied in primary cultures of differentiated cerebellar granule cells and astrocytes. After a 2-h pulse with [C3 - 3H]sphingosine at different doses (0.1-200 nmol/mg of cell protein), both cell types efficiently incorporated the long chain base ; the percentage of cellular [3H]sphingosine over total label incorporation was extremely low at sphingosine doses of <10 nmol/mg of cell protein and increased at higher doses. Most of the [3H]sphingosine taken up underwent metabolic processing by N -acylation, 1-phosphorylation, and degradation (assessed as 3H2O released in the medium). The metabolic processing of exogenous sphingosine was extremely efficient in both cells, granule cells and astrocytes being able to metabolize, respectively, an amount of sphingosine up to 80- and 300-fold the cellular content of this long chain base in 2 h. At the different doses, the prevailing metabolic route of sphingosine was different. At lower doses and in a wide dose range, the major metabolic fate of sphingosine was N -acylation. With increasing doses, there was first increased sphingosine degradation and then increased levels of sphingosine-1-phosphate. The data demonstrate that, in neurons and astrocytes, the metabolic machinery devoted to sphingosine processing is different, astrocytes possessing an overall higher capacity to synthesize the bioactive compounds ceramide and sphingosine-1-phosphate. [source] Mathematical modeling of 13C label incorporation of the TCA cycle: The concept of composite precursor functionJOURNAL OF NEUROSCIENCE RESEARCH, Issue 15 2007Kai Uffmann Abstract A novel approach for the mathematical modeling of 13C label incorporation into amino acids via the TCA cycle that eliminates the explicit calculation of the labeling of the TCA cycle intermediates is described, resulting in one differential equation per measurable time course of labeled amino acid. The equations demonstrate that both glutamate C4 and C3 labeling depend in a predictible manner on both transmitochondrial exchange rate, VX, and TCA cycle rate, VTCA. For example, glutamate C4 labeling alone does not provide any information on either VX or VTCA but rather a composite "flux". Interestingly, glutamate C3 simultaneously receives label not only from pyruvate C3 but also from glutamate C4, described by composite precursor functions that depend in a probabilistic way on the ratio of VX to VTCA: An initial rate of labeling of glutamate C3 (or C2) being close to zero is indicative of a high VX/VTCA. The derived analytical solution of these equations shows that, when the labeling of the precursor pool pyruvate reaches steady state quickly compared with the turnover rate of the measured amino acids, instantaneous labeling can be assumed for pyruvate. The derived analytical solution has acceptable errors compared with experimental uncertainty, thus obviating precise knowledge on the labeling kinetics of the precursor. In conclusion, a substantial reformulation of the modeling of label flow via the TCA cycle turnover into the amino acids is presented in the current study. This approach allows one to determine metabolic rates by fitting explicit mathematical functions to measured time courses. © 2007 Wiley-Liss, Inc. [source] In vivo 13C magnetic resonance spectroscopy of human brain on a clinical 3 T scanner using [2- 13C]glucose infusion and low-power stochastic decouplingMAGNETIC RESONANCE IN MEDICINE, Issue 3 2009Shizhe Li Abstract This study presents the detection of [2- 13C]glucose metabolism in the carboxylic/amide region in the human brain, and demonstrates that the cerebral metabolism of [2- 13C]glucose can be studied in human subjects in the presence of severe hardware constraints of widely available 3 T clinical scanners and with low-power stochastic decoupling. In the carboxylic/amide region of human brain, the primary products of 13C label incorporation from [2- 13C]glucose into glutamate, glutamine, aspartate, ,-aminobutyric acid, and N-acetylaspartate were detected. Unlike the commonly used alkanyl region where lipid signals spread over a broad frequency range, the carboxylic carbon signal of lipids was found to be confined to a narrow range centered at 172.5 ppm and present no spectral interference in the absence of lipid suppression. Comparison using phantoms shows that stochastic decoupling is far superior to the commonly used WALTZ sequence at very low decoupling power at 3 T. It was found that glutamine C1 and C5 can be decoupled using stochastic decoupling at 2.2 W, although glutamine protons span a frequency range of ,700 Hz. Detailed specific absorption rate analysis was also performed using finite difference time domain numerical simulation. Magn Reson Med, 2009. © 2009 Wiley-Liss, Inc. [source] Quantification of brain glycogen concentration and turnover through localized 13C NMR of both the C1 and C6 resonancesNMR IN BIOMEDICINE, Issue 3 2010Ruud B. van Heeswijk Abstract We have recently shown that at isotopic steady state 13C NMR can provide a direct measurement of glycogen concentration changes, but that the turnover of glycogen was not accessible with this protocol. The aim of the present study was to design, implement and apply a novel dual-tracer infusion protocol to simultaneously measure glycogen concentration and turnover. After reaching isotopic steady state for glycogen C1 using [1- 13C] glucose administration, [1,6- 13C2] glucose was infused such that isotopic steady state was maintained at the C1 position, but the C6 position reflected 13C label incorporation. To overcome the large chemical shift displacement error between the C1 and C6 resonances of glycogen, we implemented 2D gradient based localization using the Fourier series window approach, in conjunction with time-domain analysis of the resulting FIDs using jMRUI. The glycogen concentration of 5.1,±,1.6,mM measured from the C1 position was in excellent agreement with concomitant biochemical determinations. Glycogen turnover measured from the rate of label incorporation into the C6 position of glycogen in the , -chloralose anesthetized rat was 0.7,µmol/g/h. Copyright © 2009 John Wiley & Sons, Ltd. [source] | |||||||||||||