Untargeted Metabolomic Analysis of Rat Neuroblastoma Cells as a Model System to Study the Biochemical Effects of the Acute Administration of Methamphetamine

Abstract

Methamphetamine is an illicit psychostimulant drug that is linked to a number of diseases of the nervous system. The downstream biochemical effects of its primary mechanisms are not well understood, and the objective of this study was to investigate whether untargeted metabolomic analysis of an in vitro model could generate data relevant to what is already known about this drug. Rat B50 neuroblastoma cells were treated with 1 mM methamphetamine for 48 h, and both intracellular and extracellular metabolites were profiled using gas chromatography–mass spectrometry. Principal component analysis of the data identified 35 metabolites that contributed most to the difference in metabolite profiles. Of these metabolites, the most notable changes were in amino acids, with significant increases observed in glutamate, aspartate and methionine, and decreases in phenylalanine and serine. The data demonstrated that glutamate release and, subsequently, excitotoxicity and oxidative stress were important in the response of the neuronal cell to methamphetamine. Following this, the cells appeared to engage amino acid-based mechanisms to reduce glutamate levels. The potential of untargeted metabolomic analysis has been highlighted, as it has generated biochemically relevant data and identified pathways significantly affected by methamphetamine. This combination of technologies has clear uses as a model for the study of neuronal toxicology.

Metabolomics profiling of methamphetamine addicted human serum and three rat brain areas

“…Earlier studies demonstrated that METH administration could perturb amino acid metabolism. 13 By using metabolomic analysis of rat serum 13 and neuroblastoma cells, 41 amino acids were the most notable metabolites affected by METH, including glutamate, aspartate, methionine, phenylalanine and serine. Summarizing the results of METH associated metabolomics profiling, 13 amino acids were decreased in the serum of rat model except glutamate, 12 but glutamine, ornithine, histidine, asparagine, glutamate, taurine, and citrulline were increased in the serum of human METH abusers.…”

The Potential Regulatory Network of Glutamate Metabolic Pathway Disturbance in Chinese Han Withdrawal Methamphetamine Abusers

“…This study also found that the expression of the DLG2 gene was up-regulated in WMA subjects. There was a long-term cognitive impairment in methamphetamine addicted populations ( Crocker et al, 2014 ), as well as dysfunction of the glutamate system ( Maker et al, 2018 ). The DLG2 gene may play an important role in the development, plasticity, and stability of glutamatergic synapses and may further affect the cognitive function of WMA patients.…”

Toxicometabolomics: Small Molecules to Answer Big Toxicological Questions

“…A broad diversity of cellular models, including tumorigenic and nontumorigenic immortalized cell lines [ 36 , 37 , 38 ], primary cells obtained from different tissues [ 39 , 40 ], and stem cells [ 41 , 42 ] have been used in in vitro metabolomic studies, access to which is facilitated through cell culture biobanks, such as the American Type Culture Centre (ATCC, ) [ 36 ]. Immortalized cell lines offer several advantages over primary and stem cells, as they are economical and highly available, are easy to handle, can be kept in culture for longer periods of time, provide an unlimited and pure population of cells with a stable phenotype that guarantee reproducible results, and circumvent ethical concerns associated with the use of animal and/or human samples [ 29 , 31 , 43 ].…”