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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-7999836 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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