Parallel metabolomics and lipidomics enables the comprehensive study of mouse brain regional metabolite and lipid patterns
Graphical abstract
Section snippets
Author contribution
Huaxu YuNathaniel VillanuevaThibault BittarEric ArsenaultBenoit LabontéTao Huan
Materials and methods
Overall workflow. The schematic analytical workflow is presented in Fig. 1. Brain tissue was collected and preserved according to standard operating procedure (SOP) optimized to assure sample quality. The collected tissue sample was then mixed with methanol (MeOH) and sonicated for 30 min for tissue homogenization. Subsequently, methyl tert-butyl ether (MTBE) followed by water (H2O) was added to separate the homogenized solution into two layers, in which the upper layer was enriched with
Tissue homogenization and dual extraction
An appropriate tissue homogenization method should be able to efficiently break down tissue and release its metabolome content for omics-level profiling. Considering that the brain is a soft tissue and smaller brain regions are limited in tissue amount (0.2 mg of protein content), we developed a homogenization method based on sonication in an ice bath. Briefly, the brain tissue was first mixed with 270 μL methanol then sonicated in an ice bath for 30 min. During the homogenization process, we
Conclusion
Metabolites and lipids play an essential role in modulating the activity of neuronal populations across brain regions. In this work, we developed a robust and sensitive analytical workflow, requiring less than 0.2 mg of protein content, to compare metabolomic and lipidomic profiles across brain regions and provide unique information about localized metabolic activity. In our proof-of-principle, over 5000 metabolic and 6000 lipid features were detected from each brain region. Our quantitative
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
This study was funded by University of British Columbia Start-up Grant (F18-03001), Canada Foundation for Innovation [CFI 38159]. UBC Support for Teams to Advance Interdisciplinary Research Award [F19-05720], New Frontiers in Research Fund/Exploration [NFRFE-2019-00789], National Science and Engineering Research Council (NSERC) Discovery Grant [RGPIN-2020-04895], and NSERC Discovery Launch Supplement [DGECR-2020-00189]. BL holds a Sentinelle Nord Research Chair, is supported by a NARSAD young
References (49)
- et al.
Opposite molecular signatures of depression in men and women
Biol. Psychiatr.
(2018) - et al.
A systems approach identifies networks and genes linking sleep and stress: implications for neuropsychiatric disorders
Cell Rep.
(2015) - et al.
Molecular adaptations underlying susceptibility and resistance to social defeat in brain reward regions
Cell
(2007) - et al.
Shared transcriptional signatures in major depressive disorder and mouse chronic stress models
Biol. Psychiatr.
(2020) - et al.
Ketamine and imipramine reverse transcriptional signatures of susceptibility and induce resilience-specific gene expression profiles
Biol. Psychiatr.
(2017) - et al.
Integrative functional genomic analyses implicate specific molecular pathways and circuits in autism
Cell
(2013) - et al.
Integrated systems approach identifies genetic nodes and networks in late-onset Alzheimer’s disease
Cell
(2013) - et al.
Metabolomics studies in brain tissue: a review
J. Pharmaceut. Biomed. Anal.
(2016) - et al.
Region-specific metabolic alterations in the brain of the APP/PS1 transgenic mice of Alzheimer’s disease
Biochim. Biophys. Acta (BBA) - Mol. Basis Dis.
(2014) - et al.
Small molecule MALDI MS imaging: current technologies and future challenges
Methods
(2016)