{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002335","ANALYSIS_ID":"AN003814","VERSION":"1","CREATED_ON":"November 2, 2022, 6:41 am"},

"PROJECT":{"PROJECT_TITLE":"Parkinsonism mutations in DNAJC6 cause lipid defects and neurodegeneration that are rescued by Synj1","PROJECT_SUMMARY":"Recent evidence links dysfunctional lipid metabolism to the pathogenesis of Parkinson’s disease, but the mechanisms are not resolved. Here, we generated a new Drosophila knock-in model of DNAJC6/Auxilin and find that the pathogenic mutation causes synaptic dysfunction, neurological defects and neurodegeneration, as well as specific lipid metabolism alterations. In these mutants, membrane lipids containing long-chain polyunsaturated fatty acids, including phosphatidylinositol lipid species that are key for synaptic vesicle recycling and organelle function, are reduced. Overexpression of another protein mutated in Parkinson’s disease, Synaptojanin-1, known to bind and metabolize specific phosphoinositides, rescues the DNAJC6/Auxilin lipid alterations, the neuronal function defects and neurodegeneration. Our work reveals a functional relation between two proteins mutated in Parkinsonism and implicates deregulated phosphoinositide metabolism in the maintenance of neuronal integrity and neuronal survival.","INSTITUTE":"VIB-KU Leuven","DEPARTMENT":"Department of Neurosciences","LABORATORY":"Laboratory of Neuronal Communication - Dr. Patrik Verstreken","LAST_NAME":"Jacquemyn","FIRST_NAME":"Julie","ADDRESS":"Medical Sciences Building 7-25, Edmonton, Alberta, T6H 0L2, Canada","EMAIL":"jacquemynjulie@ualberta.ca","PHONE":"5873409325"},

"STUDY":{"STUDY_TITLE":"Pathogenic Auxilin mutations affect lipids that are critical to Synaptojanin function","STUDY_SUMMARY":"Mass spectrometry was performed on heads of 15DO flies homogenized in 100 µl D-PBS (Dulbecco’s phosphate-buffered saline without Mg2+ and Ca2+) by Lipotype. Fifteen fly heads were pooled from three independent crosses for each analysis, and mass spectrometry was performed on n=7 for wild-type, n≥4 analyses for dAuxWT/WT, dAuxRG/RG, dAuxWT/F956x and dAuxRG/F956x and n=3 for dAuxWT/F956x and dAuxRG/F956x overexpressing Synj.","INSTITUTE":"VIB-KU Leuven","LAST_NAME":"Jacquemyn","FIRST_NAME":"Julie","ADDRESS":"Medical Sciences Building 7-25, Edmonton, Alberta, T6H 0L2, Canada","EMAIL":"jacquemynjulie@ualberta.ca","PHONE":"+1 (587) 3409325"},

"SUBJECT":{"SUBJECT_TYPE":"Invertebrate","SUBJECT_SPECIES":"Drosophila melanogaster","TAXONOMY_ID":"7227","AGE_OR_AGE_RANGE":"15 day old flies","GENDER":"Male and female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"S1",
"Factors":{"Genotype":"AuWT/WT"}
},
{
"Subject ID":"-",
"Sample ID":"S2",
"Factors":{"Genotype":"AuWT/WT"}
},
{
"Subject ID":"-",
"Sample ID":"S3",
"Factors":{"Genotype":"AuWT/WT"}
},
{
"Subject ID":"-",
"Sample ID":"S4",
"Factors":{"Genotype":"AuWT/WT"}
},
{
"Subject ID":"-",
"Sample ID":"S5",
"Factors":{"Genotype":"AuxRG/RG"}
},
{
"Subject ID":"-",
"Sample ID":"S6",
"Factors":{"Genotype":"AuxRG/RG"}
},
{
"Subject ID":"-",
"Sample ID":"S7",
"Factors":{"Genotype":"AuxRG/RG"}
},
{
"Subject ID":"-",
"Sample ID":"S8",
"Factors":{"Genotype":"AuxRG/RG"}
},
{
"Subject ID":"-",
"Sample ID":"S9",
"Factors":{"Genotype":"AuxRG/RG"}
},
{
"Subject ID":"-",
"Sample ID":"S10",
"Factors":{"Genotype":"AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S11",
"Factors":{"Genotype":"AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S12",
"Factors":{"Genotype":"AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S13",
"Factors":{"Genotype":"AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S14",
"Factors":{"Genotype":"AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S15",
"Factors":{"Genotype":"AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S16",
"Factors":{"Genotype":"AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S17",
"Factors":{"Genotype":"AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S18",
"Factors":{"Genotype":"AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S19",
"Factors":{"Genotype":"AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S20",
"Factors":{"Genotype":"UAS-synj-AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S21",
"Factors":{"Genotype":"UAS-synj-AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S22",
"Factors":{"Genotype":"UAS-synj-AuxWT/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S23",
"Factors":{"Genotype":"UAS-synj-AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S24",
"Factors":{"Genotype":"UAS-synj-AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S25",
"Factors":{"Genotype":"UAS-synj-AuxRG/F956x"}
},
{
"Subject ID":"-",
"Sample ID":"S26",
"Factors":{"Genotype":"w1118"}
},
{
"Subject ID":"-",
"Sample ID":"S27",
"Factors":{"Genotype":"w1118"}
},
{
"Subject ID":"-",
"Sample ID":"S28",
"Factors":{"Genotype":"w1118"}
},
{
"Subject ID":"-",
"Sample ID":"S29",
"Factors":{"Genotype":"w1118"}
},
{
"Subject ID":"-",
"Sample ID":"S30",
"Factors":{"Genotype":"w1118"}
},
{
"Subject ID":"-",
"Sample ID":"S31",
"Factors":{"Genotype":"w1118"}
},
{
"Subject ID":"-",
"Sample ID":"S32",
"Factors":{"Genotype":"w1118"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Heads of 15DO flies were homogenized in 100 µl D-PBS (Dulbecco’s phosphate-buffered saline without Mg2+ and Ca2+) and stored at -80°C until processed for MS analysis by Lipotype. Fifteen fly heads were pooled from three independent crosses for each analysis, and mass spectrometry was performed on n=7 for wild-type, n≥4 analyses for dAuxWT/WT, dAuxRG/RG, dAuxWT/F956x and dAuxRG/F956x and n=3 for dAuxWT/F956x and dAuxRG/F956x overexpressing Synj.","SAMPLE_TYPE":"Insect tissue","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"In this study we did not apply any additional treatment(s) to the samples."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Lipids were extracted using chloroform and methanol (1). Prior to lipid extraction, samples were spiked with lipid class-specific internal standards. After drying and resuspending in an appropriate MS acquisition mixture, lipid extracts were infused directly in QExactive mass spectrometer (Thermo Fisher Scientific) with TriVersa NanoMate ion source (Advion Biosciences). Samples are analyzed in both positive and negative ion modes, with MS resolution Rm/z=200=280000 and MSMS resolution Rm/z=200=17500, in a single acquisition. Sample preparations and analysis were done by Lipotype. (1) Sampaio, J. L. et al. Membrane lipidome of an epithelial cell line. Proceedings of the National Academy of Sciences 108, 1903–1907 (2011)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"None (Direct infusion)","INSTRUMENT_NAME":"none","COLUMN_NAME":"none"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Samples are analyzed in both positive and negative ion modes, with MS resolution Rm/z=200=280000 and MSMS resolution Rm/z=200=17500, in a single acquisition. Acquired data was processed using lipid identification software based on LipidXplorer (1). Data post-processing and normalization were performed by Lipotype using a developed data management system. Data analysis was performed using GraphPad Prism software 9.3.1. Pmol values (obtained by MS) of individual lipid species were transformed into a fraction of the total PA, DAG, PI, PC, PE, and PS lipids in the sample. (1)Herzog, R. et al. Lipidxplorer: A software for consensual cross-platform lipidomics. PLoS ONE 7, e29851 (2012)."},

"MS_METABOLITE_DATA":{
"Units":"pMol",

"Data":[{"Metabolite":"DAG 30:0;0","S1":"0.00","S2":"3.30","S3":"10.82","S4":"4.52","S5":"7.57","S6":"3.28","S7":"9.65","S8":"10.84","S9":"9.84","S10":"0.00","S11":"2.99","S12":"10.38","S13":"10.04","S14":"18.01","S15":"5.83","S16":"1.77","S17":"14.54","S18":"10.26","S19":"12.17","S20":"11.68","S21":"0.00","S22":"0.00","S23":"13.02","S24":"8.38","S25":"9.23","S26":"0.00","S27":"0.00","S28":"1.57","S29":"0.00","S30":"9.51","S31":"4.82","S32":"7.51"},{"Metabolite":"DAG 30:1;0","S1":"54.02","S2":"39.51","S3":"201.34","S4":"42.06","S5":"63.66","S6":"29.88","S7":"125.97","S8":"136.62","S9":"181.53","S10":"60.18","S11":"34.27","S12":"251.54","S13":"202.39","S14":"333.86","S15":"57.47","S16":"17.46","S17":"61.14","S18":"74.07","S19":"108.26","S20":"243.45","S21":"320.86","S22":"318.31","S23":"134.28","S24":"90.87","S25":"100.84","S26":"22.05","S27":"29.10","S28":"16.64","S29":"16.62","S30":"224.54","S31":"111.83","S32":"256.60"},{"Metabolite":"DAG 30:2;0","S1":"14.47","S2":"7.96","S3":"35.23","S4":"8.59","S5":"17.99","S6":"4.53","S7":"22.37","S8":"33.30","S9":"43.67","S10":"21.24","S11":"9.16","S12":"71.28","S13":"49.13","S14":"80.79","S15":"23.62","S16":"5.16","S17":"18.03","S18":"21.17","S19":"29.00","S20":"53.79","S21":"68.51","S22":"82.78","S23":"46.22","S24":"37.83","S25":"36.29","S26":"8.25","S27":"9.90","S28":"4.16","S29":"4.51","S30":"60.14","S31":"32.23","S32":"68.83"},{"Metabolite":"DAG 31:0;0","S1":"0.00","S2":"0.00","S3":"0.00","S4":"3.48","S5":"5.43","S6":"0.00","S7":"0.00","S8":"0.00","S9":"0.00","S10":"0.00","S11":"0.00","S12":"0.00","S13":"0.00","S14":"0.00","S15":"2.08","S16":"0.00","S17":"14.67","S18":"0.00","S19":"3.90","S20":"0.00","S21":"0.00","S22":"0.00","S23":"10.16","S24":"6.41","S25":"7.93","S26":"0.00","S27":"0.00","S28":"1.42","S29":"0.00","S30":"0.00","S31":"0.00","S32":"0.00"},{"Metabolite":"DAG 31:1;0","S1":"5.25","S2":"7.98","S3":"24.25","S4":"16.59","S5":"10.50","S6":"4.38","S7":"25.49","S8":"21.24","S9":"26.64","S10":"0.00","S11":"5.13","S12":"27.60","S13":"21.90","S14":"27.18","S15":"10.31","S16":"2.47","S17":"16.87","S18":"12.98","S19":"23.37","S20":"20.88","S21":"25.92","S22":"26.66","S23":"57.49","S24":"37.03","S25":"42.99","S26":"2.94","S27":"0.00","S28":"2.90","S29":"3.72","S30":"23.60","S31":"20.10","S32":"24.77"},{"Metabolite":"DAG 31:2;0","S1":"0.00","S2":"0.00","S3":"0.00","S4":"4.67","S5":"0.00","S6":"0.00","S7":"0.00","S8":"0.00","S9":"0.00","S10":"0.00","S11":"0.00","S12":"0.00","S13":"0.00","S14":"0.00","S15":"2.44","S16":"0.00","S17":"3.51","S18":"0.00","S19":"0.00","S20":"0.00","S21":"0.00","S22":"0.00","S23":"11.77","S24":"7.88","S25":"7.13","S26":"0.00","S27":"0.00","S28":"0.00","S29":"0.00","S30":"0.00","S31":"0.00","S32":"0.00"},{"Metabolite":"DAG 31:3;0","S1":"0.00","S2":"0.00","S3":"0.00","S4":"2.32","S5":"0.00","S6":"0.00","S7":"0.00","S8":"0.00","S9":"0.00","S10":"0.00","S11":"0.00","S12":"0.00","S13":"0.00","S14":"0.00","S15":"0.00","S16":"0.00","S17":"0.00","S18":"0.00","S19":"0.00","S20":"0.00","S21":"0.00","S22":"0.00","S23":"0.00","S24":"0.00","S25":"0.00","S26":"0.00","S27":"0.00","S28":"0.00","S29":"0.00","S30":"0.00","S31":"0.00","S32":"0.00"},{"Metabolite":"DAG 32:0;0","S1":"0.00","S2":"0.00","S3":"0.00","S4":"0.00","S5":"4.09","S6":"0.00","S7":"0.00","S8":"0.00","S9":"0.00","S10":"0.00","S11":"0.00","S12":"0.00","S13":"0.00","S14":"0.00","S15":"2.09","S16":"0.00","S17":"12.81","S18":"0.00","S19":"0.00","S20":"0.00","S21":"0.00","S22":"0.00","S23":"0.00","S24":"0.00","S25":"0.00","S26":"0.00","S27":"0.00","S28":"0.00","S29":"2.28","S30":"0.00","S31":"0.00","S32":"0.00"},{"Metabolite":"DAG 32:1;0","S1":"39.05","S2":"20.13","S3":"109.55","S4":"22.31","S5":"30.08","S6":"14.32","S7":"85.57","S8":"76.40","S9":"101.94","S10":"41.68","S11":"15.73","S12":"170.06","S13":"114.21","S14":"240.06","S15":"28.81","S16":"6.52","S17":"37.65","S18":"44.47","S19":"75.92","S20":"122.24","S21":"168.35","S22":"193.37","S23":"81.32","S24":"46.29","S25":"59.95","S26":"15.72","S27":"23.68","S28":"8.76","S29":"7.13","S30":"168.71","S31":"69.34","S32":"164.02"},{"Metabolite":"DAG 32:2;0","S1":"36.71","S2":"19.70","S3":"70.03","S4":"17.47","S5":"33.11","S6":"8.67","S7":"56.47","S8":"60.69","S9":"83.84","S10":"45.63","S11":"13.30","S12":"153.93","S13":"96.68","S14":"199.88","S15":"32.63","S16":"6.33","S17":"32.17","S18":"40.10","S19":"59.55","S20":"108.00","S21":"112.01","S22":"164.50","S23":"67.30","S24":"52.67","S25":"46.82","S26":"16.57","S27":"25.28","S28":"7.29","S29":"7.98","S30":"158.55","S31":"63.42","S32":"136.04"},{"Metabolite":"DAG 32:3;0","S1":"0.00","S2":"0.00","S3":"0.00","S4":"4.38","S5":"0.00","S6":"0.00","S7":"0.00","S8":"0.00","S9":"5.28","S10":"5.87","S11":"0.00","S12":"8.63","S13":"5.84","S14":"0.00","S15":"0.00","S16":"0.00","S17":"0.00","S18":"0.00","S19":"0.00","S20":"0.00","S21":"0.00","S22":"13.73","S23":"0.00","S24":"3.03","S25":"0.00","S26":"2.50","S27":"3.30","S28":"0.00","S29":"0.00","S30":"11.74","S31":"7.07","S32":"13.62"},{"Metabolite":"DAG 33:0;0","S1":"0.00","S2":"0.00","S3":"0.00","S4":"0.00","S5":"0.00","S6":"0.00","S7":"0.00","S8":"0.00","S9":"0.00","S10":"0.00","S11":"0.00","S12":"0.00","S13":"0.00","S14":"0.00","S15":"0.00","S16":"0.00","S17":"14.25","S18":"0.00","S19":"0.00","S20":"0.00","S21":"0.00","S22":"0.00","S23":"0.00","S24":"0.00","S25":"0.00","S26":"0.00","S27":"0.00","S28":"0.00","S29":"0.00","S30":"0.00","S31":"0.00","S32":"0.00"},{"Metabolite":"DAG 33:1;0","S1":"0.00","S2":"3.84","S3":"14.13","S4":"11.54","S5":"7.60","S6":"0.00","S7":"14.75","S8":"14.49","S9":"19.27","S10":"0.00","S11":"2.57","S12":"20.25","S13":"14.88","S14":"22.18","S15":"5.16","S16":"1.35","S17":"16.02","S18":"10.42","S19":"20.27","S20":"0.00","S21":"0.00","S22":"13.16","S23":"36.90","S24":"26.85","S25":"28.81","S26":"0.00","S27":"0.00","S28":"1.87","S29":"0.00","S30":"16.85","S31":"11.69","S32":"19.62"},{"Metabolite":"DAG 33:2;0","S1":"0.00","S2":"0.00","S3":"7.96","S4":"7.04","S5":"3.16","S6":"0.00","S7":"6.41","S8":"5.71","S9":"8.10","S10":"0.00","S11":"0.00","S12":"8.45","S13":"6.66","S14":"0.00","S15":"2.63","S16":"0.00","S17":"5.72","S18":"4.11","S19":"5.71","S20":"0.00","S21":"0.00","S22":"0.00","S23":"16.78","S24":"10.30","S25":"12.91","S26":"0.00","S27":"0.00","S28":"0.00","S29":"0.00","S30":"9.79","S31":"7.38","S32":"12.99"},{"Metabolite":"DAG 34:0;0","S1":"0.00","S2":"2.15","S3":"5.48","S4":"0.00","S5":"0.00","S6":"1.70","S7":"5.61","S8":"0.00","S9":"0.00","S10":"0.00","S11":"1.87","S12":"0.00","S13":"0.00","S14":"0.00","S15":"0.00","S16":"1.30","S17":"10.94","S18":"5.41","S19":"0.00","S20":"4.62","S21":"0.00","S22":"0.00","S23":"6.39","S24":"4.79","S25":"5.45","S26":"0.00","S27":"0.00","S28":"1.71","S29":"1.48","S30":"7.40","S31":"0.00","S32":"7.25"},{"Metabolite":"DAG 34:1;0","S1":"17.53","S2":"6.49","S3":"36.40","S4":"7.92","S5":"12.45","S6":"4.69","S7":"41.91","S8":"34.68","S9":"44.53","S10":"22.38","S11":"4.26","S12":"65.72","S13":"41.61","S14":"94.49","S15":"15.94","S16":"2.36","S17":"22.62","S18":"26.14","S19":"46.27","S20":"32.20","S21":"45.14","S22":"61.36","S23":"39.33","S24":"22.32","S25":"28.37","S26":"7.20","S27":"9.94","S28":"3.15","S29":"1.83","S30":"61.50","S31":"23.02","S32":"52.43"},{"Metabolite":"DAG 34:2;0","S1":"24.89","S2":"10.07","S3":"46.66","S4":"12.19","S5":"17.43","S6":"5.23","S7":"39.89","S8":"35.70","S9":"49.56","S10":"27.05","S11":"8.02","S12":"90.81","S13":"54.24","S14":"115.43","S15":"18.43","S16":"3.75","S17":"24.72","S18":"31.79","S19":"51.03","S20":"53.21","S21":"56.37","S22":"79.51","S23":"44.72","S24":"29.10","S25":"35.10","S26":"13.13","S27":"16.70","S28":"4.62","S29":"3.63","S30":"100.83","S31":"42.19","S32":"86.04"},{"Metabolite":"DAG 34:3;0","S1":"10.86","S2":"3.37","S3":"12.69","S4":"3.88","S5":"6.74","S6":"0.00","S7":"8.18","S8":"9.79","S9":"10.19","S10":"14.41","S11":"0.00","S12":"25.43","S13":"14.37","S14":"34.50","S15":"4.85","S16":"0.00","S17":"6.33","S18":"8.82","S19":"9.40","S20":"17.32","S21":"26.67","S22":"36.67","S23":"9.56","S24":"8.25","S25":"5.84","S26":"8.49","S27":"11.30","S28":"2.20","S29":"2.35","S30":"31.84","S31":"14.71","S32":"27.66"},{"Metabolite":"DAG 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