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Quantification of bulk lipid species in human platelets and their thrombin-induced release
Lipids play a central role in platelet physiology. Changes in the lipidome have already been described for basal and activated platelets. However, quantitative lipidomic data of platelet activation, including the released complex lipids, are unavailable. Here we describe an easy-to-use protocol base...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10105721/ https://www.ncbi.nlm.nih.gov/pubmed/37061580 http://dx.doi.org/10.1038/s41598-023-33076-4 |
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author | Heimerl, Susanne Höring, Marcus Kopczynski, Dominik Sigruener, Alexander Hart, Christina Burkhardt, Ralph Black, Anne Ahrends, Robert Liebisch, Gerhard |
author_facet | Heimerl, Susanne Höring, Marcus Kopczynski, Dominik Sigruener, Alexander Hart, Christina Burkhardt, Ralph Black, Anne Ahrends, Robert Liebisch, Gerhard |
author_sort | Heimerl, Susanne |
collection | PubMed |
description | Lipids play a central role in platelet physiology. Changes in the lipidome have already been described for basal and activated platelets. However, quantitative lipidomic data of platelet activation, including the released complex lipids, are unavailable. Here we describe an easy-to-use protocol based on flow-injection mass spectrometry for the quantitative analysis of bulk lipid species in basal and activated human platelets and their lipid release after thrombin activation. We provide lipid species concentrations of 12 healthy human donors, including cholesteryl ester (CE), ceramide (Cer), free cholesterol (FC), hexosylceramide (HexCer), lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), sphingomyelin (SM) and triglycerides (TG). The assay exhibited good technical repeatability (CVs < 5% for major lipid species in platelets). Except for CE and TG, the inter-donor variability of the majority of lipid species concentrations in platelets was < 30% CV. Balancing of concentrations revealed the generation of LPC and loss of TG. Changes in lipid species concentrations indicate phospholipase-mediated release of arachidonic acid mainly from PC, PI, and PE but not from PS. Thrombin induced lipid release was mainly composed of FC, PS, PC, LPC, CE, and TG. The similarity of the released lipidome with that of plasma implicates that lipid release may originate from the open-canalicular system (OCS). The repository of lipid species concentrations determined with this standardized platelet release assay contribute to elucidating the physiological role of platelet lipids and provide a basis for investigating the platelet lipidome in patients with hemorrhagic or thrombotic disorders. |
format | Online Article Text |
id | pubmed-10105721 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-101057212023-04-17 Quantification of bulk lipid species in human platelets and their thrombin-induced release Heimerl, Susanne Höring, Marcus Kopczynski, Dominik Sigruener, Alexander Hart, Christina Burkhardt, Ralph Black, Anne Ahrends, Robert Liebisch, Gerhard Sci Rep Article Lipids play a central role in platelet physiology. Changes in the lipidome have already been described for basal and activated platelets. However, quantitative lipidomic data of platelet activation, including the released complex lipids, are unavailable. Here we describe an easy-to-use protocol based on flow-injection mass spectrometry for the quantitative analysis of bulk lipid species in basal and activated human platelets and their lipid release after thrombin activation. We provide lipid species concentrations of 12 healthy human donors, including cholesteryl ester (CE), ceramide (Cer), free cholesterol (FC), hexosylceramide (HexCer), lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), sphingomyelin (SM) and triglycerides (TG). The assay exhibited good technical repeatability (CVs < 5% for major lipid species in platelets). Except for CE and TG, the inter-donor variability of the majority of lipid species concentrations in platelets was < 30% CV. Balancing of concentrations revealed the generation of LPC and loss of TG. Changes in lipid species concentrations indicate phospholipase-mediated release of arachidonic acid mainly from PC, PI, and PE but not from PS. Thrombin induced lipid release was mainly composed of FC, PS, PC, LPC, CE, and TG. The similarity of the released lipidome with that of plasma implicates that lipid release may originate from the open-canalicular system (OCS). The repository of lipid species concentrations determined with this standardized platelet release assay contribute to elucidating the physiological role of platelet lipids and provide a basis for investigating the platelet lipidome in patients with hemorrhagic or thrombotic disorders. Nature Publishing Group UK 2023-04-15 /pmc/articles/PMC10105721/ /pubmed/37061580 http://dx.doi.org/10.1038/s41598-023-33076-4 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Heimerl, Susanne Höring, Marcus Kopczynski, Dominik Sigruener, Alexander Hart, Christina Burkhardt, Ralph Black, Anne Ahrends, Robert Liebisch, Gerhard Quantification of bulk lipid species in human platelets and their thrombin-induced release |
title | Quantification of bulk lipid species in human platelets and their thrombin-induced release |
title_full | Quantification of bulk lipid species in human platelets and their thrombin-induced release |
title_fullStr | Quantification of bulk lipid species in human platelets and their thrombin-induced release |
title_full_unstemmed | Quantification of bulk lipid species in human platelets and their thrombin-induced release |
title_short | Quantification of bulk lipid species in human platelets and their thrombin-induced release |
title_sort | quantification of bulk lipid species in human platelets and their thrombin-induced release |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10105721/ https://www.ncbi.nlm.nih.gov/pubmed/37061580 http://dx.doi.org/10.1038/s41598-023-33076-4 |
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