Analyzing Mass Spectrometry Imaging Data of 13C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos

The combination of 13C-isotopic labeling and mass spectrometry imaging (MSI) offers an approach to analyze metabolic flux in situ. However, combining isotopic labeling and MSI presents technical challenges ranging from sample preparation, label incorporation, data collection, and analysis. Isotopic...

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Bibliographic Details
Published in:Metabolites 2021-03, Vol.11 (3), p.148
Main Authors: Romsdahl, Trevor B., Kambhampati, Shrikaar, Koley, Somnath, Yadav, Umesh P., Alonso, Ana Paula, Allen, Doug K., Chapman, Kent D.
Format: Article
Language:English
Subjects:
13C-labeling
BASIC BIOLOGICAL SCIENCES
Camelina sativa
Carbon
Cotyledons
Data collection
Embryos
Experiments
Fatty acids
Glucose
Glycerol
INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Lasers
Lecithin
lipidomics
Lipids
MALDI (matrix assisted laser desorption/ionization)
Mass spectrometry
mass spectrometry imaging
Mass spectroscopy
Metabolic flux
Metabolism
Metabolites
metabolomics
oilseed
Oilseeds
Phosphatidylcholine
Phospholipids
Scientific imaging
Seeds
Thlaspi arvense
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Summary:The combination of 13C-isotopic labeling and mass spectrometry imaging (MSI) offers an approach to analyze metabolic flux in situ. However, combining isotopic labeling and MSI presents technical challenges ranging from sample preparation, label incorporation, data collection, and analysis. Isotopic labeling and MSI individually create large, complex data sets, and this is compounded when both methods are combined. Therefore, analyzing isotopically labeled MSI data requires streamlined procedures to support biologically meaningful interpretations. Using currently available software and techniques, here we describe a workflow to analyze 13C-labeled isotopologues of the membrane lipid and storage oil lipid intermediate―phosphatidylcholine (PC). Our results with embryos of the oilseed crops, Camelina sativa and Thlaspi arvense (pennycress), demonstrated greater 13C-isotopic labeling in the cotyledons of developing embryos compared with the embryonic axis. Greater isotopic enrichment in PC molecular species with more saturated and longer chain fatty acids suggest different flux patterns related to fatty acid desaturation and elongation pathways. The ability to evaluate MSI data of isotopically labeled plant embryos will facilitate the potential to investigate spatial aspects of metabolic flux in situ.
ISSN:2218-1989
2218-1989
DOI:10.3390/metabo11030148