{"database":"MetaboLights","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Tabular":["ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/m_MTBLS1952_LC-MS_R2_metabolite_profiling_v2_maf_pubchem.tsv","ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/m_MTBLS1952_LC-MS___metabolite_profiling_v2_maf.tsv"],"Xlsx":["ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/MTBLS1952.xlsx"],"Txt":["ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/a_MTBLS1952_LC-MS___metabolite_profiling.txt","ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/i_Investigation.txt","ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/LC and MS info from 2015 lipid mediators injections - UPDATED.txt","ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/LC and Ms info from 2015 lipid mediators injections.txt","ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/s_MTBLS1952.txt"],"Wiff":["ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/Lipid Mediators Panel A+B (Mutch, Bazinet, Leo) Re-inject 08MAY2015-Bazinet.wiff","ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952/Lipid Mediators Panel A+B (Mutch, Bazinet, Leo) Re-inject 08MAY2015-Curve.wiff"],"Other":["ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1952"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":{"citationCount":0,"reanalysisCount":0,"viewCount":0,"searchCount":0},"additional":{"submitter":["Agnes Nadjar"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS -"],"mass_spec_protocol":["
Mass spectrometry was performed as previously described[1].
Briefly, the ABSciex QTRAP5500 mass spectrometer was operated in negative electrospray ionization mode with a source temperature setting of 600 °C and a voltage setting of 4,500 V. The precursors to product ion mass transitions were obtained through scheduled multiple reaction monitoring.
Ref:
[1] Lin LE, Chen CT, Hildebrand KD, Liu Z, Hopperton KE, Bazinet RP. Chronic dietary n-6 PUFA deprivation leads to conservation of arachidonic acid and more rapid loss of DHA in rat brain phospholipids. J Lipid Res. 2015 Feb;56(2):390-402. doi:10.1194/jlr.M055590. PMID:25477531.
"],"chromatography_protocol":["LC/MS/MS was performed using a 1290 UHPLC System (Agilent Technologies, Santa Clara, CA) and a QTRAP5500 Mass Spectrometer (ABSciex, Framingham, MA). The chromatography was done at a 600 l/min flow rate on a Zorbax SB-Phenyl column (Agilent Technologies; 3.0 x 50 mm, 3.5 μm). The gradient started at 80% water and, over 9 min, ramped up to 100% acetonitrile.
"],"publication":["Essential omega-3 fatty acids tune microglial phagocytosis of synaptic elements in the developing brain. 10.1038/s41467-020-19861-z."],"submitter_affiliation":["University of Bordeaux"],"Organism":["Mus musculus","blank","Reference compound"],"technology_type":["mass spectrometry"],"disease":[""],"extraction_protocol":["Pooled microglia extracted from 3-4 brains per sample were lysed in 15% methanol and stored at -80 degree C prior to analysis. All procedures used siliconized glassware. Samples were extracted in the dark on ice to prevent auto-oxidation. Samples were extracted as previously reported[1].
Briefly, 1 ng of internal standard mixture was added to a 250 mg aliquot of each sample. External ARA, EPA (20:5n-3), and DHA standards were prepared in a similar way. The samples were mixed for 1 min, incubated on ice for 30 min, and centrifuged at 1,000 x g for 10 min. The supernatants were collected. The pellet was resuspended in ethanol for 1 min and centrifuged again for a second extraction. The resultant ethanolic supernatants were combined with the methanolic supernatants, previously extracted. After evaporation with nitrogen gas, the supernatants were suspended in 10% ethanol, acidified to pH 3 with 1 N HCl, and triply extracted with ethyl acetate. The ethyl acetate layer was washed to neutrality with water and dried under nitrogen gas. The residues from the brain and external standard samples were reconstituted in acetonitrile-water (1:1 by volume) and transferred into the inserts of amber vials for immediate LC/MS/MS analysis.
Ref:
[1] Lin LE, Chen CT, Hildebrand KD, Liu Z, Hopperton KE, Bazinet RP. Chronic dietary n-6 PUFA deprivation leads to conservation of arachidonic acid and more rapid loss of DHA in rat brain phospholipids. J Lipid Res. 2015 Feb;56(2):390-402. doi:10.1194/jlr.M055590. PMID:25477531.
"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS1952"],"author":["Agnes Nadjar. agnes.nadjar@u-bordeaux.fr."],"data_transformation_protocol":["Data analysis for microglia lipid analysis was performed as previously described[1].
Briefly, composite standards of lipid metabolites (natural or deuterated; Cayman Chemicals Co., Ann Ar-bor, MI) were diluted in ethanol from stock solutions to perform an 8-point calibration curve (0.05 to 5 ng). Internal standard mixtures in ethanol were added to both the composite standards and the samples prior to extraction.
Quantitative analysis was performed by Analyst 1.5.2 Software (AB-Sciex). The area ratios of the integrated peaks (natural to deuter-ated standard) were plotted against the standard curves for quantification. The limit of quantification was 0.025 ng per sample, and values between 0.005 ng and 0.025 ng were considered semiquantitative.
Ref:
[1] Lin LE, Chen CT, Hildebrand KD, Liu Z, Hopperton KE, Bazinet RP. Chronic dietary n-6 PUFA deprivation leads to conservation of arachidonic acid and more rapid loss of DHA in rat brain phospholipids. J Lipid Res. 2015 Feb;56(2):390-402. doi:10.1194/jlr.M055590. PMID:25477531.
"],"study_factor":["Diet"],"submitter_email":[""],"sample_collection_protocol":["Male and female CD1 mice 8-week old were purchased from Janvier Labs (Le Gesnest St Isle, France) and were SPF-housed in temperature and humidity-controlled cages on a 12 h light/dark cycle with food and water ad libitum.
After mating, CD1 females were randomly assigned to either a n-3 deficient or a n-3 sufficient diet, both containing 5% fat, in the form of sunflower oil (rich in LA, the 'n-3 deficient diet') or a mixture of different oils of which rapeseed oil (rich in ALA, the 'n-3 sufficient diet') throughout gestation and lactation1–4 (Table 1 and 2). All experiments were performed on the offspring from n-3 deficient and n-3 sufficient dams at post-natal day (P) 21.
Lipid analyses were performed on hippocampal synaptosomes or microglia. To this aim, hippocampus or total brain were isolated from juvenile n-3 PUFA deficient vs control mice.
Synaptosomes were prepared from P21 mice hippocampus. Briefly, the 2 hippocampi of each animal were dissected and placed in 1 ml of ice-cold iso-osmolar buffer. 2 cm^3 tissue samples were homogenized with a Potter tissue grinder (900 rpm, 12 strokes). The homogenate was centrifuged at 1,000 x g for 5 min at 4 °C, the supernatant was collected in a new tube and centrifuged at 12,500 x g for 8 min at 4 °C. The supernatant was removed and the pellet resuspended with 1 ml of ice-cold iso-osmolar buffer. The discontinuous sucrose gradients were prepared in ultracentrifuge tubes with ice-cold 1.2 M sucrose solution and 0.8 M sucrose solution. The resuspended pellet was placed on top of the sucrose gradient. The centrifugation at 50,000 x g for 1 h at 4 °C provided a visible layer at the interface of sucrose gradients. This fraction (synaptosomes) was collected with a syringe. Synaptosomes were then rinsed with 100 mM pH 8.5 sodium bicarbonate buffer and stored until required.
Lipid mediators were identified by comparison with commercial standards purchased from Cayman Chemicals Co., (Ann Arbor MI, USA).
"],"metabolite_name":["PGD2","12S-HEPE","8,9-EET","8-HETE","Prostaglandin F3alpha","(+/-)-8-HEPE","(+/-)-11-HEPE","TXB3","15S-HEPE","TXB2","trioxilin B3","LTB5","LTB4","arachidonic acid","9-HETE","18-HEPE","12-HETE","Lipoxin A5","9-HEPE","15-HETE","Prostaglandin D3","5,6-EET","11,12-EET","8-Iso-PGF3alpha","8(9)-EpETE","14,15-EET","7(R)Marisin1","trioxilin A3","14-HDoHE","LXA4","PD1","DHA","17(18)-EpETE","delta(17)-6-ketoprostaglandin F1alpha","11-Dehydrothromboxane B2","5-HEPE","5-HETE","7-HDoHE","17S-HDoHE","8-iso Prostaglandin F2alpha","PGF2a","10(S),17(s)-DiHDoHE","Resolvin E1","PGE3","PGE2","17R-Resolvin","Resolvin-D1","Resolvin-D2","11-HETE","4-HDoHE","6-keto PGF1a","EPA","PGJ2"],"description_synonyms":["projections, acido graso, Cognitive Function, Disorders, acidos grasos, Under-Developed, single-organism developmental process, H-rev142, Ammon horn fields, Third-World Nation, histology, Less-Developed, Least, primary ILD specific to childhood, Less-Developed Country, element, Disorders Usually Diagnosed in Infancy, Relative, rrg, Developed Country, Abnormal retropulsion test, diseases, Formations, symptoms, Saturated Fatty Acids, Synapse, diseases and disorders, Esterified Fatty Acid, synganglion, Lower Middle Income Country, Under Developed Countries, Epidemiologic Study, Esterified, average, me75, human disease, Oxidoreductase, Esterified Fatty, Aliphatic, suprasegmental levels of nervous system, Less-Developed Countries, Developing Nation, D17Mit170, Ammons Horn, T1, Aliphatic Acids, Cognitions, Ammons horn, Functions, Habit Disturbance, papilla, signaling process, Homo sapiens disease, Mental Disorders, Third World Countries, primary interstitial lung disease specific to childhood, single organism signaling, ammon gyrus, screening, Countries, suprasegmental structures, anatomical protrusion, Child Mental Disorder, Child Mental, Ammon, pediatric interstitial lung disease, Less Developed Nations, formatio hippocampi, Low Income, Epidemiologic, lamina, flanges, Ammon Horn, Fatty acid, hippocampus proper, Fettsaeuren, Normalcy, Epidemiological Studies, Tl3, Tl2, Habit Disturbances, results, Girl, seahorse, Subiculums, Developing Nations, Lower-Middle-Income, Hippocampal, ARABIDOPSIS LIPOXYGENASE 1, Relative Risks, Diseases, shelf, Under-Developed Countries, Linoleate Oxygen Oxidoreductase, major hippocampus, atomo, mixed synapse, Ammon's Horn, Neurodevelopmental Disorders Usually Diagnosed in Infancy, atome, Phagocytoses, Subiculum, activation, child, Linoleate:oxygen 13-oxidoreductase, Acid, LOX-IMVI, Fatty Acid, Ammon's, dmlox-1, Ammon's horn, Element, Saturated Fatty Acid, Child Mental Disorders, Risk, ILD specific to childhood, shelves, atoms, Third-World Country, signs, Aliphatic Acid, projection, Children, ridge, Developing Country, Low Income Countries, disease, Neurodevelopmental, fatty acids, synaptic junction, Neurodevelopmental Disorders., Health, primal cortex, AI893619, spine, Linoleate-Oxygen, electrotonic synapse, neurodevelopmental disorder, DmelCG11335, Mental Disorder, Saturated Fatty, the brain, Lower-Middle-Income Countries, Middle Income, other disease, Esterified Fatty Acids, biological signaling, Least Developed Countries, young adult, Least Developed, lamellae, Neurodevelopmental Disorder, ammon horn, lipoxygenase 1, LOX IMVI, process of organ, Child, protrusion, lamella, Childhood or Adolescence, acides gras, CNS development, LMICs, Rrg1, CHILD, Girls, Diets, Studies, LIPOXYGENASE, disease or disorder, Imbalance, LOX, Lox, Low, Country, Third-World Nations, interstitial lung disease of childhood, Cornu Ammonis, Lipoxygenase 1, Hippocampus, Lipoxygenase 2, Normalities, Under-Developed Nations, Fatty Acids, Less Developed Countries, Mental Disorders Diagnosed in Childhood, hippocampus (Crosby), ridges, Middle Income Country, hippocampus major, paediatric interstitial lung disease, chILD, non-neoplastic, Cognitive Functions, Study, juvenile stage, Under Developed Nations, Disorder, childhood interstitial lung disease, Habit, disorder, elements, laminae, Neurodevelopmental Disorders, Epidemiological, atom, Horn, Third-World Countries, Third-World, Cornu ammonis, Saturated, Lower-Middle-Income Country, findings, cou, Lipoxygenase-2, Lipoxygenase-1, Women's Group, DmLOXL-1, anatomical process, Proper, Women Groups, disorders, Hippocampus Propers, Hippocampal Formation, medical condition, Third World Nations, encephalon, Woman, Under-Developed Nation, cognitive function, development, Fettsaeure, Lr, archipallium, Linoleate-Oxygen Oxidoreductase, Lipoxidase, Least Developed Country, Low and Middle Income Countries, condition, Epidemiological Study, atomus, Balance impairment, flange, organ process, Hippocampus Proper, Relative Risk, Nation, cornu ammonis, hippocampus proprius, Propers, Normality, Dietary, Women's Groups, Risks, Function, chILD syndrome, CNS - Brain - Hippocampus (MMHCC), acide gras, Cognitive, signalling, Low Income Country, ATLOX1, processes, process, CG11335, hippocampus, Less-Developed Nations, signalling process, children's interstitial lung disease, Bra, Less-Developed Nation, Mental Disorders Usually Diagnosed in Infancy, TSC-160, Middle Income Countries, processus, Dmloxl-1, Hippocampal Formations, Under-Developed Country, Formation"],"name_synonyms":["the brain, suprasegmental structures, Element, atoms, atomo, atomus, synganglion, elements, atome, suprasegmental levels of nervous system, Phagocytoses, atom, encephalon, Encephalon., element"],"citation_count_scaled":["0.0"],"download_count_scaled":["0.0"],"normalized_connections":["0.0"],"reanalysis_count_scaled":["0.0"],"view_count_scaled":["0.0"],"citation_count":["0"],"additional_accession":[]},"is_claimable":false,"name":"Essential omega-3 fatty acids tune microglial phagocytosis of synaptic elements in the developing brain","description":"Omega-3 fatty acids (n-3 polyunsaturated fatty acids; n-3 PUFAs) are essential for the functional maturation of the brain. Westernization of dietary habits in both developed and developing countries is accompanied by a progressive reduction in dietary intake of n-3 PUFAs. Low maternal intake of n-3 PUFAs has been linked to neurodevelopmental diseases in epidemiological studies, but the mechanisms by which a n-3 PUFA dietary imbalance affects CNS development are poorly understood. Active microglial engulfment of synaptic elements is an important process for normal brain development and altered synapse refinement is a hallmark of several neurodevelopmental disorders. Here, we identify a molecular mechanism for detrimental effects of low maternal n-3 PUFA intake on hippocampal development. Our results show that maternal dietary n-3 PUFA deficiency increases microglial phagocytosis of synaptic elements in the developing hippocampus, partly through the activation of 12/15- lipoxygenase (LOX)/12-HETE signaling, which alters neuronal morphology and affects cognition in the postnatal offspring. While women of child bearing age are at higher risk of dietary n-3 PUFA deficiency, these findings provide new insights into the mechanisms linking maternal nutrition to neurodevelopmental disorders.
","dates":{"publication":"2020-11-03","submission":"2020-07-23"},"accession":"MTBLS1952","cross_references":{"MetaboLights":["MTBLC34157","MTBLC131889","MTBLC28364","MTBLC19138","MTBLC34505","MTBLC88493","MTBLC72786","MTBLC72624","MTBLC36005","MTBLC72623","MTBLC36203","MTBLC15647","MTBLC84444","MTBLC15843","MTBLC28031","MTBLC28158","MTBLC64017","MTBLC34939","MTBLC6498","MTBLC138179","MTBLC140930","MTBLC89570","MTBLC72853","MTBLC35032","MTBLC81565","MTBLC138655","MTBLC60943","MTBLC81564","MTBLC28667","MTBLC137193","MTBLC138640","MTBLC28728","MTBLC72802","MTBLC72647","MTBLC72801","MTBLC15551","MTBLC72606","MTBLC15555","MTBLC72643","MTBLC15553","MTBLC88347","MTBLC34450","MTBLC157745","MTBLC88345","MTBLC34130","MTBLC36075","MTBLC34490","MTBLC138249","MTBLC27485","MTBLC157746","MTBLC81559","MTBLC157747","MTBLC157748"],"ChEBI":["CHEBI:28667","CHEBI:72786","CHEBI:72623","CHEBI:138640","CHEBI:84444","CHEBI:88493","CHEBI:72624","CHEBI:137193","CHEBI:34505","CHEBI:15553","CHEBI:138249","CHEBI:15555","CHEBI:34157","CHEBI:15551","CHEBI:27485","CHEBI:28728","CHEBI:60943","CHEBI:131889","CHEBI:81559","CHEBI:28364","CHEBI:72643","CHEBI:72801","CHEBI:72647","CHEBI:72802","CHEBI:15843","CHEBI:72606","CHEBI:15647","CHEBI:6498","CHEBI:88345","CHEBI:34450","CHEBI:36075","CHEBI:88347","CHEBI:34130","CHEBI:19138","CHEBI:64017","CHEBI:157747","CHEBI:157748","CHEBI:157745","CHEBI:157746","CHEBI:34490","CHEBI:34939","CHEBI:81565","CHEBI:28158","CHEBI:72853","CHEBI:28031","CHEBI:81564","CHEBI:89570","CHEBI:140930","CHEBI:35032","CHEBI:36203","CHEBI:138655","CHEBI:36005","CHEBI:138179"]}}