Ter homeostasis was assessed in the transgenic McGill-RThy1-APP rat modelTer homeostasis was assessed within the

Ter homeostasis was assessed in the transgenic McGill-RThy1-APP rat model
Ter homeostasis was assessed within the transgenic McGill-RThy1-APP rat model of AD. In these rats, accumulation of Ab oligomers seems 1 week right after birth and cognitive symptoms are apparent by three months of age. Extracellular Ab plaques start accumulating in the subiculum region at age 6 months, appear in most areas in the hippocampal formation and some locations on the cerebral cortex at age 13 months, and are located in most areas in the brain by 20 months of age.10 We’ve previously reported that changes in metabolite concentrations are readily detected by in vivo 1H NMR spectroscopy at each early and much more sophisticated age in these rats.11 In the present study, neuronal and astrocytic metabolism was studied simultaneously by injecting transgenic McGill-R-Thy1-APP rats and age-matched Caspase 2 web controls with [1-13C]glucose and [1,2-13C]acetate followed by analysis with ex vivo 1H and 13C NMR spectroscopy and high-performance liquid chromatography (HPLC). We investigated metabolic alterations inside the hippocampal formation, frontal-, entorhinal-, and retrosplenialcingulate cortices due to the fact regional hypometabolism of glucose in AD happens in brain regions like the1 Department of Neuroscience, Faculty of Medicine, Norwegian University of Science and Technology, Trondheim, Norway and 2Centre for Neural Computation, Faculty of Medicine, Kavli Institute for Systems Neuroscience, Norwegian University of Science and Technology, Trondheim, Norway. Correspondence: Professor U Sonnewald, Department of Neuroscience, Faculty of Medicine, Norwegian University of Science and Technologies, PO Box 8905, MTFS, Trondheim 7491, Norway. E-mail: ursula.sonnewaldntnu.no Received 21 August 2013; revised 18 January 2014; accepted 25 January 2014; published online 5 MarchBrain metabolism inside a rat model of AD LH Nilsen et al907 posterior cingulate cortex and the medial temporal lobe, as well as in the frontal cortex in later stages of the disease.12,13 Materials AND Approaches Materials[1-13C]glucose and [1,2-13C]acetate were bought from Cambridge Isotope Laboratories (Andover, MA, USA), deuterium oxide (D2O, 99.9 ) from CDN Isotopes (Point-Claire, Quebec, Canada), ethylene glycol from Merck (Darmstadt, Germany) and two,2-Dimethyl-2-silapentane-5-sulfonate sodium salt (DSS sodium salt) from Sigma-Aldrich (St Louis, MO, USA). All other chemicals of the purest grade had been accessible from local commercial suppliers. was collected and transferred to a brand new tube. The remaining chloroform phase was re-extracted by adding 400 mL methanol, 300 mL purified water, and 100 mL chloroform. Following centrifugation, the new methanolwater phase was pooled together with the methanolwater phase collected previously. The chloroform phase was after once more re-extracted and centrifuged, along with the methanolwater phase was pooled with these previously collected for every sample. All samples have been kept on ice whenever probable throughout the extraction procedure and stored at 801C following extraction. Following lyophilization, the samples were resuspended in 200 mL D2O, centrifuged at B3,000 g for 10 IDO2 web minutes at 41C, and 5 mL was removed in the supernatants for HPLC analysis. The supernatants have been then lyophilized twice with D2O. Concentrations of metabolites and incorporation of 13C label into metabolites in brain extracts obtained from transgenic McGill-R-Thy1-APP rats and controls have been quantified working with HPLC, 1H and 13C NMR spectroscopy. As a consequence of the little size from the entorhinal cortex, 13C NMR spectroscopy spectra with sufficient signal-to-noise ra.