{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000294","ANALYSIS_ID":"AN000471","VERSION":"1","CREATED_ON":"December 27, 2015, 4:25 pm"},

"PROJECT":{"PROJECT_TITLE":"Mechanisms of Metabolic Cycles in Diapausing Flesh Fly by Metabolomics Approach","PROJECT_TYPE":"Time course","PROJECT_SUMMARY":"Insects use diapause, a programmed period of dormancy, to avoid stressful times of the year and to exploit seasonal times of resource availability. Because most diapausing insects do not feed, they must live off their body reserves for several months and the proper use of metabolic reserves is critical for surviving diapause and performing after diapause termination. Across multiple insects, metabolic depression during diapause has been associated with a switch from aerobic metabolism to facultative anaerobic metabolism, despite insects not suffering environmental oxygen limitation. While metabolic rates are depressed during diapause overall to save energy, some insects show regular cyclical bouts of higher metabolic activity during diapause. The functional importance of these metabolic cycles and the mechanisms underlying these cycles are still unknown, but they may be critical for properly maintaining the balance between energy states and purge the accumulation of anaerobic metabolic byproducts. In the present study, we will test the hypothesis that periodic cycles of increased metabolism during insect diapause are associated with both regenerating organismal energetic states, particularly ATP that may decline during metabolic depression, and for purging metabolites associated with anaerobic metabolism. We will use a combination of non-targeted uHPLC-MS/MS metabolomics and targeted NMR-spectroscopy to identify and quantify metabolites that are altered during the cycles in diapausing pupae of the flesh fly, Sarcophaga crassipalpis. This work will allow us to propose specific biochemical and cellular hypotheses for the regulation of cyclic releases from metabolic depression in diapausing insects. Our work may not only reveal the physiological mechanisms regulating metabolic cycles during diapause in flesh fly, but also provide insight to understand the regulation of similar metabolic cycles in mammalian hibernators (i.e., periodic arousal), and also provide insights into how these cycles could be exploited to disrupt the diapause of insect pests.","INSTITUTE":"The Institute of Food and Agricultural Sciences","DEPARTMENT":"Department of Entomology and Nematology","LAST_NAME":"Hahn","FIRST_NAME":"Daniel","ADDRESS":"P.O. Box 110620 Gainesville FL 32611-0620","EMAIL":"dahahn@ufl.edu","PHONE":"352-273-3968"},

"STUDY":{"STUDY_TITLE":"Mechanisms of Metabolic Cycles in Diapausing Flesh Fly by Metabolomics Approach","STUDY_TYPE":"time course","STUDY_SUMMARY":"Insects use diapause, a programmed period of dormancy, to avoid stressful times of the year and to exploit seasonal times of resource availability. Because most diapausing insects do not feed, they must live off their body reserves for several months and the proper use of metabolic reserves is critical for surviving diapause and performing after diapause termination. Across multiple insects, metabolic depression during diapause has been associated with a switch from aerobic metabolism to facultative anaerobic metabolism, despite insects not suffering environmental oxygen limitation. While metabolic rates are depressed during diapause overall to save energy, some insects show regular cyclical bouts of higher metabolic activity during diapause. The functional importance of these metabolic cycles and the mechanisms underlying these cycles are still unknown, but they may be critical for properly maintaining the balance between energy states and purge the accumulation of anaerobic metabolic byproducts. In the present study, we will test the hypothesis that periodic cycles of increased metabolism during insect diapause are associated with both regenerating organismal energetic states, particularly ATP that may decline during metabolic depression, and for purging metabolites associated with anaerobic metabolism. We will use a combination of non-targeted uHPLC-MS/MS metabolomics and targeted NMR-spectroscopy to identify and quantify metabolites that are altered during the cycles in diapausing pupae of the flesh fly, Sarcophaga crassipalpis. This work will allow us to propose specific biochemical and cellular hypotheses for the regulation of cyclic releases from metabolic depression in diapausing insects. Our work may not only reveal the physiological mechanisms regulating metabolic cycles during diapause in flesh fly, but also provide insight to understand the regulation of similar metabolic cycles in mammalian hibernators (i.e., periodic arousal), and also provide insights into how these cycles could be exploited to disrupt the diapause of insect pests.","INSTITUTE":"University of Florida","DEPARTMENT":"SECIM","LAST_NAME":"Chen","FIRST_NAME":"Chao","ADDRESS":"Department of Entomology and Nematology, Bldg. 970, 1881 Natural Area Dr., Gainesville, FL 32611","EMAIL":"chaochenjxau@126.com","PHONE":"352-273-3949","NUM_GROUPS":"5","TOTAL_SUBJECTS":"45"},

"SUBJECT":{"SUBJECT_TYPE":"Insect","SUBJECT_SPECIES":"Sarcophaga crassipalpis","GENOTYPE_STRAIN":"Lab colony","AGE_OR_AGE_RANGE":"15-25 day pupae","WEIGHT_OR_WEIGHT_RANGE":"110-130 mg","ANIMAL_ANIMAL_SUPPLIER":"lab colony","ANIMAL_HOUSING":"sterilite containers","ANIMAL_LIGHT_CYCLE":"10:14 L:D","ANIMAL_FEED":"beef liver","ANIMAL_INCLUSION_CRITERIA":"must be in diapause"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"B1P101",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B1P102",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B1P105",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B1P203",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B1P202",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B1P205",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B1P301",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B1P305",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B1P306",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B1P402",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B1P403",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B1P406",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B1P504",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B1P505",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B1P506",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B2P101",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B2P102",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B2P103",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B2P201",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B2P202",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"BEP203",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B2P301",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B2P302",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B2P304",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B2P401",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B2P402",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B2P403",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B2P502",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B2P503",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B2P504",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B3P103",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B3P105",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B3P106",
"Factors":{"Phase":"ED"}
},
{
"Subject ID":"-",
"Sample ID":"B3P202",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B3P204",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B3P205",
"Factors":{"Phase":"LD"}
},
{
"Subject ID":"-",
"Sample ID":"B3P302",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B3P303",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B3P304",
"Factors":{"Phase":"AD"}
},
{
"Subject ID":"-",
"Sample ID":"B3P402",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B3P403",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B3P404",
"Factors":{"Phase":"IBA"}
},
{
"Subject ID":"-",
"Sample ID":"B3P503",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B3P504",
"Factors":{"Phase":"EN"}
},
{
"Subject ID":"-",
"Sample ID":"B3P506",
"Factors":{"Phase":"EN"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Pupae were sampled at the appropriate time point and flash frozen in liquid nitrogen. Pupae were stored at -80C until extraction.","COLLECTION_PROTOCOL_COMMENTS":"Samples were homogenized in ice cold 80% methanol and centrifuged. Supernatant was retained and stored at -80C until analysis.","SAMPLE_TYPE":"whole fly","COLLECTION_METHOD":"flash freezing in liquid nitrogen","COLLECTION_LOCATION":"Hahn Lab, 3137 Steinmetz","COLLECTION_FREQUENCY":"Daily","COLLECTION_TIME":"see treatments","VOLUMEORAMOUNT_COLLECTED":"one pupa per sample","STORAGE_CONDITIONS":"-80C","COLLECTION_VIALS":"1.5 ml microcentrifuge tubes","STORAGE_VIALS":"1.5 ml microcentrifuge tubes","COLLECTION_TUBE_TEMP":"room temperature"},

"TREATMENT":{"TREATMENT_SUMMARY":"ED=Early metabolic depression (48h) LD=Late metabolic depression (144h) AD=Arousing from metabolic depression (6h) IBA=Interbout arousal (15h) EN=Entering metabolic depression (30h)"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"This sample set is FleshFly tissue. The samples were received dried and frozen. The samples were reconstituted in 1 mL 90:10 Water:Acetonitrile then diluted 10x."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Scientific-Dionex Ultimate 3000","COLUMN_NAME":"ACE Excel 2 C18-PFP (100 x 2.1mm, 2um)","METHODS_FILENAME":"Metabolomics_LCMSProtocol.pdf"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"Metabolomics_LCMSProtocol.pdf"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap Thermo Scientific- Q Exactive"},

"MS_METABOLITE_DATA":{
"Units":"Peak height",

"Data":[{"Metabolite":"(+/-)-6-Acetonyldihydrosanguinarine","B3P504":"1.72E+07","B3P105":"1.98E+07","B3P204":"2.08E+07","B3P506":"2.18E+07","B1P506":"2.17E+07","B2P304":"2.22E+07","B3P303":"1.96E+07","B2P103":"1.77E+07","B3P202":"1.94E+07","B2P502":"2.26E+07","B1P305":"1.86E+07","B2P504":"2.32E+07","B2P202":"2.43E+07","B1P306":"2.56E+07","B3P402":"1.72E+07","B1P105":"2.03E+07","B3P106":"2.4E+07","B3P403":"2.19E+07","B2P402":"2.13E+07","B1P403":"2.11E+07","B2P102":"2.04E+07","BEP203":"2.08E+07","B1P505":"1.85E+07","B1P402":"1.86E+07","B3P503":"1.97E+07","B2P201":"1.63E+07","B1P102":"1.17E+07","B2P101":"2.18E+07","B1P202":"1.78E+07","B1P301":"1.77E+07","B1P101":"1.81E+07","B2P301":"1.85E+07","B1P504":"2.27E+07","B2P503":"1.96E+07","B3P404":"2.06E+07","B3P304":"2.13E+07","B1P203":"1.7E+07","B2P403":"1.83E+07","B3P103":"1.97E+07","B2P302":"1.74E+07","B1P205":"2.5E+07","B2P401":"1.78E+07","B3P302":"2.18E+07","B3P205":"2.41E+07","B1P406":"2.03E+07"},{"Metabolite":"(4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexaenoic 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}