Protocols for Metabolomic Analyses to Assess Changes in Central Carbon Metabolism from Neural Tissue

Metabolomics has been successfully applied to study neurological and neurodegenerative disorders, including Parkinson's disease for the following purposes: (1) identifying potential biomarkers of onset and disease progression; (2) identifying novel mechanisms of disease progression; and (3) assessing treatment prognosis and outcome. Reproducible and efficient extraction of metabolites is imperative to the success of any metabolomics investigation. Unlike other OMICS techniques, the composition of the metabolome can be negatively impacted by the preparation, processing, and handling of these samples. The proper choice of data collection, preprocessing, and processing protocols are similarly important to the design of an effective metabolomics experiment. Likewise, correctly applying univariate and multivariate statistical methods is essential for providing biologically relevant insights. In this chapter, we have outlined a detailed metabolomics workflow that addresses these issues. A step-by-step protocol from preparing neuronal cells and metabolomic tissue samples to their metabolic analyses using nuclear magnetic resonance, mass spectrometry, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), and chemometrics is presented.