Effects of hypercapnia / ischemia and dissection on the rat brain metabolome.

It is known that brain energy metabolites such as ATP are quickly depleted during postmortem ischemia; however, a comprehensive assessment on the effects of preceding hypercapnia/ischemia and the dissection process on the larger brain metabolome remains lacking. This study sought to address this unknown by measuring aqueous metabolites impacted by hypercapnia/ischemia and brain dissection using Nuclear Magnetic Resonance. Metabolites were measured in rats subjected to 1) high energy head-focused microwave irradiation (control group); 2) CO2-induced hypercapnia/ischemia followed by immediate microwave irradiation; 3) CO2 followed by decapitation and then microwave irradiation ∼6.4 min later, to simulate a postmortem interval equivalent to typical dissection times; and 4) CO2-induced hypercapnia/ischemia followed by dissection within ∼6 min (no microwave fixation) to test the effects of brain dissection on the metabolome. Compared to control rats subjected to head-focused microwave irradiation, concentrations of high-energy phosphate metabolites and glucose were significantly reduced, while ß-hydroxybutyrate and lactate were increased in rats subjected to all other treatments. Several amino acids and neurotransmitters (GABA) increased by hypercapnia/ischemia and dissection. Sugar donors involved in glycosylation decreased and nucleotides decreased or increased following hypercapnia/ischemia and dissection. sn-Glycero-3-phosphocholine decreased and its choline byproduct increased in all groups relative to controls, indicating postmortem changes in lipid turnover. Antioxidants increased following hypercapnia/ischemia but decreased to control levels following dissection. This study demonstrates substantial post-mortem changes in brain energy and glycosylation pathways, as well as protein, nucleotide, neurotransmitter, lipid, and antioxidant turnover due to hypercapnia/ischemia and dissection. Changes in phosphate donors, glycosylation and amino acids reflect post-translational modification and protein degradation processes that persist post-mortem. Microwave irradiation is necessary for accurately capturing in vivo brain metabolite concentrations.

Untargeted metabolomic analysis of plasma from relapsing-remitting multiple sclerosis patients reveals changes in metabolites associated with structural changes in brain.

BACKGROUND: Changes in peripheral blood amino acids have been noted in Relapse Remitting Multiple Sclerosis (RRMS), suggesting their potential diagnostic value in anticipating disease progression. OBJECTIVE: The present study sought to comprehensively assess the plasma metabolome, including amino acids, of RRMS patient and unaffected controls, to identify potential biomarkers of RRMS disease pathogenesis. METHODS: Untargeted NMR metabolomics was performed on plasma from 28 RRMS patients and 18 unaffected controls to test the hypothesis that metabolomic markers are altered in RRMS patients in association with lesion load, brain atrophy and cognitive performance. RESULTS: There were no significant differences between RRMS and controls in age, sex and total brain volume. Brain fractional volumes of gray matter, white matter, thalamus and parenchyma as well as multiple neurocognitive scores were significantly lower in RRMS patients compared to unaffected controls. Concentrations of nine plasma metabolites (arginine, isoleucine, citrate, serine, phenylalanine, methionine, asparagine, histidine, myo-inositol) were significantly lower in RRMS patients compared to controls. Plasma arginine concentrations were positively correlated with T1 holes and white matter lesions, and plasma methionine concentrations were positively correlated with T1 holes, but not white matter lesions. Serine was negatively correlated with performance on the Brief Visuospatial Memory Test in controls but not RRMS patients. CONCLUSIONS: The identified disturbances in metabolite concentrations might be developed as new markers of neuroanatomical vulnerability in RRMS, should the findings be reproduced in larger cohort studies.