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β - N -Methylamino- l -alanine (BMAA) perturbs alanine, aspartate and glutamate metabolism pathways in human neuroblastoma cells as determined by metabolic profiling

View Article: PubMed Central - PubMed

ABSTRACT

β-Methylamino-l-alanine (BMAA) is a non-proteinogenic amino acid that induces long-term cognitive deficits, as well as an increased neurodegeneration and intracellular fibril formation in the hippocampus of adult rodents following short-time neonatal exposure and in vervet monkey brain following long-term exposure. It has also been proposed to be involved in the etiology of neurodegenerative disease in humans. The aim of this study was to identify metabolic effects not related to excitotoxicity or oxidative stress in human neuroblastoma SH-SY5Y cells. The effects of BMAA (50, 250, 1000 µM) for 24 h on cells differentiated with retinoic acid were studied. Samples were analyzed using LC–MS and NMR spectroscopy to detect altered intracellular polar metabolites. The analysis performed, followed by multivariate pattern recognition techniques, revealed significant perturbations in protein biosynthesis, amino acid metabolism pathways and citrate cycle. Of specific interest were the BMAA-induced alterations in alanine, aspartate and glutamate metabolism and as well as alterations in various neurotransmitters/neuromodulators such as GABA and taurine. The results indicate that BMAA can interfere with metabolic pathways involved in neurotransmission in human neuroblastoma cells.

No MeSH data available.


Representative NMR spectra obtained from the pooled QC sample dissolved in 0.7 ml phosphate buffered D2O (150 mM, pD 7). Chemical shifts are expressed relative to the DSS resonance set at 0.00 ppm
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Fig3: Representative NMR spectra obtained from the pooled QC sample dissolved in 0.7 ml phosphate buffered D2O (150 mM, pD 7). Chemical shifts are expressed relative to the DSS resonance set at 0.00 ppm

Mentions: The profiling experiments based on NMR-spectroscopy resulted in information rich spectra, i.e. containing a plentitude of signals of varying intensity; an example is given in Fig. 3. In general, all spectra were of high intensity. However, this also results in severe overlap in certain spectral regions (see for example region at δ 4.5–2.0 p.p.m in Fig. 3). Consequently, unambiguous metabolite identifications were elusive, but some plausible annotations are presented at the end of this section. Generated OPLS-DA models demonstrated adequate group differentiation for the high (1.0 mM) and medium (250 µM) BMAA concentration samples as compared to untreated control cells. However, the experiments using a low BMAA (50 µM) concentration resulted in models with a poor predictive power (Q2 < 0.4). Spectral bins were selected from S-plots according to the criteria described in materials and methods; these spectral bins were subsequently tested for significance by ANOVA with parametric post hoc Tukey’s tests. Briefly, forty-six spectral bins were significantly altered of which only a few could be tentatively assigned due to overlap of resonances. Briefly, following exposure to the high concentration of BMAA there was an enrichment of the following tentatively assigned metabolites: l-aspartate, taurine and l-serine. Moreover, following exposure to the medium concentration of BMAA there was an enrichment of the following tentatively assigned metabolites: hypotaurine, taurine and histamine, whereas cytidine monophosphate was reduced.Fig. 3


β - N -Methylamino- l -alanine (BMAA) perturbs alanine, aspartate and glutamate metabolism pathways in human neuroblastoma cells as determined by metabolic profiling
Representative NMR spectra obtained from the pooled QC sample dissolved in 0.7 ml phosphate buffered D2O (150 mM, pD 7). Chemical shifts are expressed relative to the DSS resonance set at 0.00 ppm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383692&req=5

Fig3: Representative NMR spectra obtained from the pooled QC sample dissolved in 0.7 ml phosphate buffered D2O (150 mM, pD 7). Chemical shifts are expressed relative to the DSS resonance set at 0.00 ppm
Mentions: The profiling experiments based on NMR-spectroscopy resulted in information rich spectra, i.e. containing a plentitude of signals of varying intensity; an example is given in Fig. 3. In general, all spectra were of high intensity. However, this also results in severe overlap in certain spectral regions (see for example region at δ 4.5–2.0 p.p.m in Fig. 3). Consequently, unambiguous metabolite identifications were elusive, but some plausible annotations are presented at the end of this section. Generated OPLS-DA models demonstrated adequate group differentiation for the high (1.0 mM) and medium (250 µM) BMAA concentration samples as compared to untreated control cells. However, the experiments using a low BMAA (50 µM) concentration resulted in models with a poor predictive power (Q2 < 0.4). Spectral bins were selected from S-plots according to the criteria described in materials and methods; these spectral bins were subsequently tested for significance by ANOVA with parametric post hoc Tukey’s tests. Briefly, forty-six spectral bins were significantly altered of which only a few could be tentatively assigned due to overlap of resonances. Briefly, following exposure to the high concentration of BMAA there was an enrichment of the following tentatively assigned metabolites: l-aspartate, taurine and l-serine. Moreover, following exposure to the medium concentration of BMAA there was an enrichment of the following tentatively assigned metabolites: hypotaurine, taurine and histamine, whereas cytidine monophosphate was reduced.Fig. 3

View Article: PubMed Central - PubMed

ABSTRACT

&beta;-Methylamino-l-alanine (BMAA) is a non-proteinogenic amino acid that induces long-term cognitive deficits, as well as an increased neurodegeneration and intracellular fibril formation in the hippocampus of adult rodents following short-time neonatal exposure and in vervet monkey brain following long-term exposure. It has also been proposed to be involved in the etiology of neurodegenerative disease in humans. The aim of this study was to identify metabolic effects not related to excitotoxicity or oxidative stress in human neuroblastoma SH-SY5Y cells. The effects of BMAA (50, 250, 1000&nbsp;&micro;M) for 24&nbsp;h on cells differentiated with retinoic acid were studied. Samples were analyzed using LC&ndash;MS and NMR spectroscopy to detect altered intracellular polar metabolites. The analysis performed, followed by multivariate pattern recognition techniques, revealed significant perturbations in protein biosynthesis, amino acid metabolism pathways and citrate cycle. Of specific interest were the BMAA-induced alterations in alanine, aspartate and glutamate metabolism and as well as alterations in various neurotransmitters/neuromodulators such as GABA and taurine. The results indicate that BMAA can interfere with metabolic pathways involved in neurotransmission in human neuroblastoma cells.

No MeSH data available.