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

The combination of (13)C-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. Isotopi...

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Autores principales: Romsdahl, Trevor B., Kambhampati, Shrikaar, Koley, Somnath, Yadav, Umesh P., Alonso, Ana Paula, Allen, Doug K., Chapman, Kent D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999836/
https://www.ncbi.nlm.nih.gov/pubmed/33806402
http://dx.doi.org/10.3390/metabo11030148
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author Romsdahl, Trevor B.
Kambhampati, Shrikaar
Koley, Somnath
Yadav, Umesh P.
Alonso, Ana Paula
Allen, Doug K.
Chapman, Kent D.
author_facet Romsdahl, Trevor B.
Kambhampati, Shrikaar
Koley, Somnath
Yadav, Umesh P.
Alonso, Ana Paula
Allen, Doug K.
Chapman, Kent D.
author_sort Romsdahl, Trevor B.
collection PubMed
description The combination of (13)C-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 (13)C-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 (13)C-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.
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spelling pubmed-79998362021-03-28 Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos Romsdahl, Trevor B. Kambhampati, Shrikaar Koley, Somnath Yadav, Umesh P. Alonso, Ana Paula Allen, Doug K. Chapman, Kent D. Metabolites Article The combination of (13)C-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 (13)C-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 (13)C-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. MDPI 2021-03-04 /pmc/articles/PMC7999836/ /pubmed/33806402 http://dx.doi.org/10.3390/metabo11030148 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Romsdahl, Trevor B.
Kambhampati, Shrikaar
Koley, Somnath
Yadav, Umesh P.
Alonso, Ana Paula
Allen, Doug K.
Chapman, Kent D.
Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos
title Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos
title_full Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos
title_fullStr Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos
title_full_unstemmed Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos
title_short Analyzing Mass Spectrometry Imaging Data of (13)C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos
title_sort analyzing mass spectrometry imaging data of (13)c-labeled phospholipids in camelina sativa and thlaspi arvense (pennycress) embryos
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999836/
https://www.ncbi.nlm.nih.gov/pubmed/33806402
http://dx.doi.org/10.3390/metabo11030148
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