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. 2024 Feb 5;43(4):533–567. doi: 10.1038/s44318-024-00030-7

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© The Author(s) 2024

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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) Schematic workflow for Free Fatty Acid (FFA) extraction from different brain regions including cortex (CX), ventral hippocampus (VH), amygdala (AM), dorsal hippocampus (DH) and cerebellum (CB), and subsequent derivatization using FFAST, and LC-MS/MS analysis. For each brain region, FFAs were extracted from the two different experimental conditions (instrumental or context control mice) and labelled separately at the carboxy-terminus using FFAST for subsequent multiplexing LC-MS/MS analysis, together with FFAST-labelled internal standards (as per Narayana et al, 2015 (Narayana et al, 2015)). This workflow was replicated 6 times, to establish FFA abundance in each of the 6 mice used for each experimental condition. (B) Bar plots showing the profile of 19 FFAs obtained by FFAST LC-MS/MS across 6 brains of 12-month-old control vs instrumentally conditioned DDHD2^+/+ vs DDHD2^−/− mice with analytes shown by acyl chain composition. Data information: In (B), the significance of the difference between each group (n = 6 biological replicates) as determined by unpaired t-test with Holm–Sidak post hoc correction is indicated by asterisks **p < 0.01, ***p < 0.001, ns = not significant. Error bars represent the cumulative standard error of the mean (SEM) for all groups and parameters. Source data are available online for this figure.