Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 408, Issue 13, Pages 3453-3474Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-016-9421-3
Keywords
IR-MALDESI; Proteomics; Lipidomics; Mass spectrometry imaging; Embryonic brain
Funding
- NIH [R01NS089795, R01GM087964]
- NIH/NCSU Molecular Biotechnology Training Grant [5T32GM00-8776-08]
- DAAD (German Academic Exchange Service)
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Quantitative methods for detection of biological molecules are needed more than ever before in the emerging age of omics and big data. Here, we provide an integrated approach for systematic analysis of the lipidome in tissue. To test our approach in a biological context, we utilized brain tissue selectively deficient for the transcription factor Specificity Protein 2 (Sp2). Conditional deletion of Sp2 in the mouse cerebral cortex results in developmental deficiencies including disruption of lipid metabolism. Silver (Ag) cationization was implemented for infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) to enhance the ion abundances for olefinic lipids, as these have been linked to regulation by Sp2. Combining Ag-doped and conventional IR-MALDESI imaging, this approach was extended to IR-MALDESI imaging of embryonic mouse brains. Further, our imaging technique was combined with bottom-up shotgun proteomic LC-MS/MS analysis and western blot for comparing Sp2 conditional knockout (Sp2-cKO) and wild-type (WT) cortices of tissue sections. This provided an integrated omics dataset which revealed many specific changes to fundamental cellular processes and biosynthetic pathways. In particular, step-specific altered abundances of nucleotides, lipids, and associated proteins were observed in the cerebral cortices of Sp2-cKO embryos.
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