{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003271","ANALYSIS_ID":"AN005357","PROJECT_ID":"PR002030","VERSION":"1","CREATED_ON":"June 18, 2024, 9:59 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic changes in mouse tissues following modulation of serine through the diet","PROJECT_SUMMARY":"We analyzed metabolites in the retina, choroid/RPE, and plasma from WT and PHGDH heterozygous mice that were fed either a control or serine/glycine-deprived diet. Additionally, we also analyzed metabolites from the retina, choroid/RPE, paw skin, and plasma from mice that were previously on either a control or serine/glycine-deprived diet and then switched back to a control or serine-supplemented diet.","INSTITUTE":"Salk Institute for Biological Studies","DEPARTMENT":"Molecular and Cell Biology Laboratory","LABORATORY":"Metallo Lab","LAST_NAME":"Lim","FIRST_NAME":"Esther","ADDRESS":"10010 N Torrey Pines Rd, La Jolla, California, 92037, USA","EMAIL":"ewlim2024@gmail.com","PHONE":"(858) 453-4100","FUNDING_SOURCE":"R01CA234245"},

"STUDY":{"STUDY_TITLE":"Impact of serine supplementation following treatment with serine/glycine-depleted diet","STUDY_SUMMARY":"We analyzed metabolites in the retina, choroid/RPE, plasma, and paw skin from mice that were previously on control or serine/glycine-depleted diet and then switched over to a control or serine-supplemented diet.","INSTITUTE":"Salk Institute for Biological Studies","DEPARTMENT":"Molecular and Cell Biology Laboratory","LABORATORY":"Metallo Lab","LAST_NAME":"Lim","FIRST_NAME":"Esther","ADDRESS":"10010 N Torrey Pines Rd","EMAIL":"ewlim2024@gmail.com","PHONE":"(858) 453-4100","NUM_GROUPS":"4","TOTAL_SUBJECTS":"20","NUM_MALES":"20"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6J","AGE_OR_AGE_RANGE":"12-16 months","WEIGHT_OR_WEIGHT_RANGE":"25-45 g","GENDER":"Male","ANIMAL_ANIMAL_SUPPLIER":"The Scripps Research Institute Vivarium","ANIMAL_HOUSING":"5/cage","ANIMAL_LIGHT_CYCLE":"12h dark/12 h light","ANIMAL_FEED":"Special diets; see group labeling","ANIMAL_WATER":"Ad libitum"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20B2",
"Factors":{"Treatment":"Paw skin  control","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20B2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20B3",
"Factors":{"Treatment":"Paw skin  control","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20B3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20B4",
"Factors":{"Treatment":"Paw skin  control","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20B4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20C2",
"Factors":{"Treatment":"Paw skin  control","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20C2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20C3",
"Factors":{"Treatment":"Paw skin  control","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20C3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20D1",
"Factors":{"Treatment":"Paw skin  -SG diet","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20D1.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20D2",
"Factors":{"Treatment":"Paw skin  -SG diet","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20D2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20D3",
"Factors":{"Treatment":"Paw skin  -SG diet","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20D3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20D4",
"Factors":{"Treatment":"Paw skin  -SG diet","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20D4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20D5",
"Factors":{"Treatment":"Paw skin  -SG diet","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20D5.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20H1",
"Factors":{"Treatment":"Paw skin  High serine rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20H1.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20H2",
"Factors":{"Treatment":"Paw skin  High serine rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20H2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20H3",
"Factors":{"Treatment":"Paw skin  High serine rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20H3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20H4",
"Factors":{"Treatment":"Paw skin  High serine rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20H4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20H5",
"Factors":{"Treatment":"Paw skin  High serine rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20H5.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20J1",
"Factors":{"Treatment":"Paw skin  Control rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20J1.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20J2",
"Factors":{"Treatment":"Paw skin  Control rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20J2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20J3",
"Factors":{"Treatment":"Paw skin  Control rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20J3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20J4",
"Factors":{"Treatment":"Paw skin  Control rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20J4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_Skin_20J5",
"Factors":{"Treatment":"Paw skin  Control rescue","Sample source":"Mouse paw skin"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_Skin_20J5.d"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Tissues were collected using Wollenberger clamps pre-cooled to the temperature of liquid nitrogen and stored at -80 C until analysis","SAMPLE_TYPE":"Skin"},

"TREATMENT":{"TREATMENT_SUMMARY":"Mice were fed control or serine/glycine-depleted (-SG) diet for 12 months. After 12 months, a subset of the mice that were on the -SG diet were switched back to either the control or supplemented diet for 4 months."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For sphingolipid extraction from retina and choroid/RPE, frozen tissue was homogenized with a ball mill (Retsch Mixer Mill MM 400) at 30 Hz for 3 min in 500 μl of −20°C methanol, 400 μl of ice-cold saline, and 100 μl of ice-cold MilliQ water, spiked with deuterated internal standards as described earlier. The mixture was then transferred into a 2-ml Eppendorf tube containing 1 ml of chloroform. The tubes were vortexed for 5 min and centrifuged at 16,000g at 4°C for 5 min. The lower organic phase was collected, and 2 μl of formic acid was added to the remaining polar phase, which was re-extracted with 1 ml of chloroform. Combined organic phases were dried and resuspended in 50 μl of 0.2% formic acid and 1 mM ammonium formate in methanol. Last, the tubes were sonicated in a bath sonicator for 10 min and spun at 16,000g for 10 min at 4°C. For plasma, 50 μl of plasma was extracted with 500 μl of −20°C methanol, 400 μl of saline, and 100 μl of water spiked with deuterated internal standards. Chloroform (1 ml) was then added to the tubes. The tubes were vortexed for 5 min and centrifuged at 16,000g at 4°C for 5 min. The lower organic phase was collected, and 2 μl of formic acid was added to the remaining polar phase, which was reextracted with 1 ml of chloroform. Combined organic phases were dried and resuspended in 100 μl of 0.2% formic acid and 1 mM ammonium formate in methanol. Next, the tubes were sonicated in a bath sonicator for 10 min and spun at 16,000g for 10 min at 4°C","PROCESSING_STORAGE_CONDITIONS":"Room temperature","EXTRACT_STORAGE":"4℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"The gradient elution program consisted of the following profile: 0 min, 82% B; 3 min, 82% B; 4 min, 90% B, 18 min, 99% B; 25 min, 99%, 27 min, 82% B, 30 min, 82% B.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 6460","COLUMN_NAME":"Spectra 3µm C8SR column (3 μm particle, 3.0 × 150 mm; Peeke Scientific)","SOLVENT_A":"100% water; 0.2% formic acid; 2 mM ammonium formate","SOLVENT_B":"100% methanol; 0.2% formic acid; 1 mM ammonium formate","FLOW_GRADIENT":"0 min, 82% B; 3 min, 82% B; 4 min, 90% B; 18 min, 99% B; 25 min, 99%; 27 min, 82% B; and 30 min, 82% B","FLOW_RATE":"0.5 mL/min","COLUMN_TEMPERATURE":"40"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent 6460 QQQ","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Software: Agilent MassHunter Sphingolipid species were analyzed by selective reaction monitoring of the transition from precursor to product ions at associated optimized collision energies, and fragmentor voltages are provided elsewhere (https://doi.org/10.1007/978-1-60761-322-0_22). The m/z values of the precursor and product ions are provided in the following format: Precursor ion -> Product ion."},

"MS_METABOLITE_DATA":{
"Units":"Normalized ion count/mg of protein",

"Data":[{"Metabolite":"Cer d18:1/14:0","LMRI_DietRev_Skin_20B2":"4.994197","LMRI_DietRev_Skin_20B3":"5.028954059","LMRI_DietRev_Skin_20B4":"5.355774369","LMRI_DietRev_Skin_20C2":"6.196536392","LMRI_DietRev_Skin_20C3":"6.672178351","LMRI_DietRev_Skin_20D1":"10.25256771","LMRI_DietRev_Skin_20D2":"5.665478688","LMRI_DietRev_Skin_20D3":"6.666832906","LMRI_DietRev_Skin_20D4":"5.461108369","LMRI_DietRev_Skin_20D5":"5.92374284","LMRI_DietRev_Skin_20H1":"6.203601384","LMRI_DietRev_Skin_20H2":"4.207017525","LMRI_DietRev_Skin_20H3":"3.724001561","LMRI_DietRev_Skin_20H4":"4.413352432","LMRI_DietRev_Skin_20H5":"3.502558531","LMRI_DietRev_Skin_20J1":"4.369686971","LMRI_DietRev_Skin_20J2":"3.808812905","LMRI_DietRev_Skin_20J3":"6.330419245","LMRI_DietRev_Skin_20J4":"5.486839622","LMRI_DietRev_Skin_20J5":"5.995304717"},{"Metabolite":"Cer d18:1/16:0","LMRI_DietRev_Skin_20B2":"56.96837461","LMRI_DietRev_Skin_20B3":"95.74873386","LMRI_DietRev_Skin_20B4":"69.55112943","LMRI_DietRev_Skin_20C2":"75.90712757","LMRI_DietRev_Skin_20C3":"58.2437605","LMRI_DietRev_Skin_20D1":"114.6467488","LMRI_DietRev_Skin_20D2":"63.98969778","LMRI_DietRev_Skin_20D3":"53.01935516","LMRI_DietRev_Skin_20D4":"61.55014003","LMRI_DietRev_Skin_20D5":"53.97434953","LMRI_DietRev_Skin_20H1":"79.46133323","LMRI_DietRev_Skin_20H2":"68.93728122","LMRI_DietRev_Skin_20H3":"55.06168141","LMRI_DietRev_Skin_20H4":"71.66780082","LMRI_DietRev_Skin_20H5":"64.61221719","LMRI_DietRev_Skin_20J1":"61.2934226","LMRI_DietRev_Skin_20J2":"75.17729608","LMRI_DietRev_Skin_20J3":"93.50676646","LMRI_DietRev_Skin_20J4":"80.49507114","LMRI_DietRev_Skin_20J5":"79.78762336"},{"Metabolite":"Cer d18:1/16:1","LMRI_DietRev_Skin_20B2":"3.814998466","LMRI_DietRev_Skin_20B3":"5.383949848","LMRI_DietRev_Skin_20B4":"5.056870728","LMRI_DietRev_Skin_20C2":"5.691347962","LMRI_DietRev_Skin_20C3":"4.13298142","LMRI_DietRev_Skin_20D1":"5.387142957","LMRI_DietRev_Skin_20D2":"4.72612649","LMRI_DietRev_Skin_20D3":"4.495674701","LMRI_DietRev_Skin_20D4":"4.110527478","LMRI_DietRev_Skin_20D5":"3.491614339","LMRI_DietRev_Skin_20H1":"6.516721408","LMRI_DietRev_Skin_20H2":"5.992798582","LMRI_DietRev_Skin_20H3":"3.882845229","LMRI_DietRev_Skin_20H4":"4.694805888","LMRI_DietRev_Skin_20H5":"4.113714586","LMRI_DietRev_Skin_20J1":"4.915310179","LMRI_DietRev_Skin_20J2":"5.277682456","LMRI_DietRev_Skin_20J3":"7.244076341","LMRI_DietRev_Skin_20J4":"6.616740689","LMRI_DietRev_Skin_20J5":"5.056399449"},{"Metabolite":"Cer d18:1/18:0","LMRI_DietRev_Skin_20B2":"25.82456136","LMRI_DietRev_Skin_20B3":"39.28971887","LMRI_DietRev_Skin_20B4":"37.52344656","LMRI_DietRev_Skin_20C2":"43.25724914","LMRI_DietRev_Skin_20C3":"32.9915431","LMRI_DietRev_Skin_20D1":"46.46917271","LMRI_DietRev_Skin_20D2":"35.98030453","LMRI_DietRev_Skin_20D3":"30.42937806","LMRI_DietRev_Skin_20D4":"36.55072608","LMRI_DietRev_Skin_20D5":"37.64625557","LMRI_DietRev_Skin_20H1":"37.61764099","LMRI_DietRev_Skin_20H2":"35.42098838","LMRI_DietRev_Skin_20H3":"32.91865959","LMRI_DietRev_Skin_20H4":"34.51264922","LMRI_DietRev_Skin_20H5":"34.66282576","LMRI_DietRev_Skin_20J1":"32.90500406","LMRI_DietRev_Skin_20J2":"37.27535879","LMRI_DietRev_Skin_20J3":"38.02889113","LMRI_DietRev_Skin_20J4":"34.09655345","LMRI_DietRev_Skin_20J5":"34.5520809"},{"Metabolite":"Cer d18:1/18:1","LMRI_DietRev_Skin_20B2":"5.242762823","LMRI_DietRev_Skin_20B3":"7.914992736","LMRI_DietRev_Skin_20B4":"8.921095518","LMRI_DietRev_Skin_20C2":"13.14292395","LMRI_DietRev_Skin_20C3":"8.462708562","LMRI_DietRev_Skin_20D1":"11.9119792","LMRI_DietRev_Skin_20D2":"9.928221686","LMRI_DietRev_Skin_20D3":"5.741614169","LMRI_DietRev_Skin_20D4":"7.179584164","LMRI_DietRev_Skin_20D5":"6.61317256","LMRI_DietRev_Skin_20H1":"17.13453114","LMRI_DietRev_Skin_20H2":"13.00393756","LMRI_DietRev_Skin_20H3":"8.535661637","LMRI_DietRev_Skin_20H4":"14.38986775","LMRI_DietRev_Skin_20H5":"13.6166728","LMRI_DietRev_Skin_20J1":"9.18293127","LMRI_DietRev_Skin_20J2":"18.65164905","LMRI_DietRev_Skin_20J3":"19.66907762","LMRI_DietRev_Skin_20J4":"19.24073753","LMRI_DietRev_Skin_20J5":"18.66129547"},{"Metabolite":"Cer d18:1/20:0","LMRI_DietRev_Skin_20B2":"69.47315483","LMRI_DietRev_Skin_20B3":"87.53096679","LMRI_DietRev_Skin_20B4":"80.68227506","LMRI_DietRev_Skin_20C2":"84.47993447","LMRI_DietRev_Skin_20C3":"63.74897727","LMRI_DietRev_Skin_20D1":"154.5680541","LMRI_DietRev_Skin_20D2":"79.14353457","LMRI_DietRev_Skin_20D3":"84.02582434","LMRI_DietRev_Skin_20D4":"62.24918933","LMRI_DietRev_Skin_20D5":"87.48645027","LMRI_DietRev_Skin_20H1":"96.09470603","LMRI_DietRev_Skin_20H2":"73.67636735","LMRI_DietRev_Skin_20H3":"55.77367382","LMRI_DietRev_Skin_20H4":"76.24093364","LMRI_DietRev_Skin_20H5":"67.98127594","LMRI_DietRev_Skin_20J1":"63.62376788","LMRI_DietRev_Skin_20J2":"64.73484814","LMRI_DietRev_Skin_20J3":"112.5254537","LMRI_DietRev_Skin_20J4":"86.6271135","LMRI_DietRev_Skin_20J5":"99.53540183"},{"Metabolite":"Cer d18:1/22:0","LMRI_DietRev_Skin_20B2":"97.90061354","LMRI_DietRev_Skin_20B3":"138.0681234","LMRI_DietRev_Skin_20B4":"148.9664501","LMRI_DietRev_Skin_20C2":"143.9696511","LMRI_DietRev_Skin_20C3":"119.9210221","LMRI_DietRev_Skin_20D1":"198.1413869","LMRI_DietRev_Skin_20D2":"148.9590866","LMRI_DietRev_Skin_20D3":"126.9292885","LMRI_DietRev_Skin_20D4":"127.781412","LMRI_DietRev_Skin_20D5":"128.9980642","LMRI_DietRev_Skin_20H1":"134.1801265","LMRI_DietRev_Skin_20H2":"122.922786","LMRI_DietRev_Skin_20H3":"86.52179983","LMRI_DietRev_Skin_20H4":"115.7915237","LMRI_DietRev_Skin_20H5":"107.2687903","LMRI_DietRev_Skin_20J1":"105.1825005","LMRI_DietRev_Skin_20J2":"123.0803457","LMRI_DietRev_Skin_20J3":"156.5689006","LMRI_DietRev_Skin_20J4":"120.0620839","LMRI_DietRev_Skin_20J5":"137.4534856"},{"Metabolite":"Cer d18:1/23:0","LMRI_DietRev_Skin_20B2":"22.07082978","LMRI_DietRev_Skin_20B3":"31.23834461","LMRI_DietRev_Skin_20B4":"34.45184521","LMRI_DietRev_Skin_20C2":"37.7616687","LMRI_DietRev_Skin_20C3":"26.52187592","LMRI_DietRev_Skin_20D1":"38.81346722","LMRI_DietRev_Skin_20D2":"34.58843476","LMRI_DietRev_Skin_20D3":"27.80957167","LMRI_DietRev_Skin_20D4":"35.82522361","LMRI_DietRev_Skin_20D5":"29.48047","LMRI_DietRev_Skin_20H1":"29.49992122","LMRI_DietRev_Skin_20H2":"27.36173125","LMRI_DietRev_Skin_20H3":"19.52498785","LMRI_DietRev_Skin_20H4":"21.62071208","LMRI_DietRev_Skin_20H5":"20.6730245","LMRI_DietRev_Skin_20J1":"22.5795209","LMRI_DietRev_Skin_20J2":"31.968999","LMRI_DietRev_Skin_20J3":"28.48879536","LMRI_DietRev_Skin_20J4":"26.52600017","LMRI_DietRev_Skin_20J5":"25.78205579"},{"Metabolite":"Cer d18:1/24:0","LMRI_DietRev_Skin_20B2":"126.9502905","LMRI_DietRev_Skin_20B3":"180.487734","LMRI_DietRev_Skin_20B4":"247.8644994","LMRI_DietRev_Skin_20C2":"244.9327811","LMRI_DietRev_Skin_20C3":"165.1872522","LMRI_DietRev_Skin_20D1":"242.3969092","LMRI_DietRev_Skin_20D2":"238.6403661","LMRI_DietRev_Skin_20D3":"128.5935482","LMRI_DietRev_Skin_20D4":"189.9150892","LMRI_DietRev_Skin_20D5":"145.121953","LMRI_DietRev_Skin_20H1":"199.3276221","LMRI_DietRev_Skin_20H2":"204.8222373","LMRI_DietRev_Skin_20H3":"113.9796516","LMRI_DietRev_Skin_20H4":"147.7996512","LMRI_DietRev_Skin_20H5":"161.4578843","LMRI_DietRev_Skin_20J1":"153.935038","LMRI_DietRev_Skin_20J2":"190.7475559","LMRI_DietRev_Skin_20J3":"179.0444127","LMRI_DietRev_Skin_20J4":"173.8828219","LMRI_DietRev_Skin_20J5":"148.4437621"},{"Metabolite":"Cer 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}

}