#METABOLOMICS WORKBENCH ZJ309_20231112_222225 DATATRACK_ID:4456 STUDY_ID:ST002981 ANALYSIS_ID:AN004900 PROJECT_ID:PR001857 VERSION 1 CREATED_ON November 16, 2023, 12:29 am #PROJECT PR:PROJECT_TITLE Developmental Neurotoxicity of Deltamethrin Exposure on Hypothalamic PR:PROJECT_TITLE Neurogenesis in Embryonic Zebrafish PR:PROJECT_SUMMARY In this study, to investigate DM effects at different stages of early life, PR:PROJECT_SUMMARY zebrafish embryos were exposed to DM just before (10-16 hpf), at the onset of PR:PROJECT_SUMMARY (16-24 hpf), at the peak of (24-36 hpf) hypothalamic neurogenesis and across PR:PROJECT_SUMMARY 10-120 hpf with different dosage levels (0, 1, 100, and 250 nM). PR:INSTITUTE College of Marine Food and Biological Engineering, Jimei University PR:LAST_NAME Gao PR:FIRST_NAME Longhua PR:ADDRESS No. 185 Yinjiang Road, Jimei District, Xiamen City, Fujian Province, China PR:EMAIL 1932076629@qq.com PR:PHONE 18718180398 #STUDY ST:STUDY_TITLE Developmental Neurotoxicity of Deltamethrin Exposure on Hypothalamic ST:STUDY_TITLE Neurogenesis in Embryonic Zebrafish ST:STUDY_SUMMARY In this study, to investigate DM effects at different stages of early life, ST:STUDY_SUMMARY zebrafish embryos were exposed to DM just before (10-16 hpf), at the onset of ST:STUDY_SUMMARY (16-24 hpf), at the peak of (24-36 hpf) hypothalamic neurogenesis and across ST:STUDY_SUMMARY 10-120 hpf with different dosage levels (0, 1, 100, and 250 nM). ST:INSTITUTE College of Marine Food and Biological Engineering, Jimei University ST:LAST_NAME Gao ST:FIRST_NAME Longhua ST:ADDRESS No. 185 Yinjiang Road, Jimei District, Xiamen City, Fujian Province, China ST:EMAIL 1932076629@qq.com ST:PHONE 18718180398 #SUBJECT SU:SUBJECT_TYPE Fish SU:SUBJECT_SPECIES Danio rerio SU:TAXONOMY_ID 7955 #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS 0_1 0_1 Treatment:Control Model=Positive/Negitive; RAW_FILE_NAME=0_1.RAW SUBJECT_SAMPLE_FACTORS 0_2 0_2 Treatment:Control Model=Positive/Negitive; RAW_FILE_NAME=0_2.RAW SUBJECT_SAMPLE_FACTORS 0_3 0_3 Treatment:Control Model=Positive/Negitive; RAW_FILE_NAME=0_3.RAW SUBJECT_SAMPLE_FACTORS 0_4 0_4 Treatment:Control Model=Positive/Negitive; RAW_FILE_NAME=0_4.RAW SUBJECT_SAMPLE_FACTORS 0_5 0_5 Treatment:Control Model=Positive/Negitive; RAW_FILE_NAME=0_5.RAW SUBJECT_SAMPLE_FACTORS 0_6 0_6 Treatment:Control Model=Positive/Negitive; RAW_FILE_NAME=0_6.RAW SUBJECT_SAMPLE_FACTORS 1n-10-16_1 1n-10-16_1 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-16_1.RAW SUBJECT_SAMPLE_FACTORS 1n-10-16_2 1n-10-16_2 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-16_2.RAW SUBJECT_SAMPLE_FACTORS 1n-10-16_3 1n-10-16_3 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-16_3.RAW SUBJECT_SAMPLE_FACTORS 1n-10-16_4 1n-10-16_4 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-16_4.RAW SUBJECT_SAMPLE_FACTORS 1n-10-16_5 1n-10-16_5 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-16_5.RAW SUBJECT_SAMPLE_FACTORS 1n-10-16_6 1n-10-16_6 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-16_6.RAW SUBJECT_SAMPLE_FACTORS 1n-16-24_1 1n-16-24_1 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-16-24_1.RAW SUBJECT_SAMPLE_FACTORS 1n-16-24_2 1n-16-24_2 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-16-24_2.RAW SUBJECT_SAMPLE_FACTORS 1n-16-24_3 1n-16-24_3 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-16-24_3.RAW SUBJECT_SAMPLE_FACTORS 1n-16-24_4 1n-16-24_4 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-16-24_4.RAW SUBJECT_SAMPLE_FACTORS 1n-16-24_5 1n-16-24_5 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-16-24_5.RAW SUBJECT_SAMPLE_FACTORS 1n-16-24_6 1n-16-24_6 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-16-24_6.RAW SUBJECT_SAMPLE_FACTORS 1n-24-36_1 1n-24-36_1 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-24-36_1.RAW SUBJECT_SAMPLE_FACTORS 1n-24-36_2 1n-24-36_2 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-24-36_2.RAW SUBJECT_SAMPLE_FACTORS 1n-24-36_3 1n-24-36_3 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-24-36_3.RAW SUBJECT_SAMPLE_FACTORS 1n-24-36_4 1n-24-36_4 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-24-36_4.RAW SUBJECT_SAMPLE_FACTORS 1n-24-36_5 1n-24-36_5 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-24-36_5.RAW SUBJECT_SAMPLE_FACTORS 1n-24-36_6 1n-24-36_6 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-24-36_6.RAW SUBJECT_SAMPLE_FACTORS 1n-10-5d_1 1n-10-5d_1 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-5d_1.RAW SUBJECT_SAMPLE_FACTORS 1n-10-5d_2 1n-10-5d_2 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-5d_2.RAW SUBJECT_SAMPLE_FACTORS 1n-10-5d_3 1n-10-5d_3 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-5d_3.RAW SUBJECT_SAMPLE_FACTORS 1n-10-5d_4 1n-10-5d_4 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-5d_4.RAW SUBJECT_SAMPLE_FACTORS 1n-10-5d_5 1n-10-5d_5 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-5d_5.RAW SUBJECT_SAMPLE_FACTORS 1n-10-5d_6 1n-10-5d_6 Treatment:1nM Model=Positive/Negitive; RAW_FILE_NAME=1n-10-5d_6.RAW SUBJECT_SAMPLE_FACTORS 100n-10-16_1 100n-10-16_1 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-16_1.RAW SUBJECT_SAMPLE_FACTORS 100n-10-16_2 100n-10-16_2 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-16_2.RAW SUBJECT_SAMPLE_FACTORS 100n-10-16_3 100n-10-16_3 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-16_3.RAW SUBJECT_SAMPLE_FACTORS 100n-10-16_4 100n-10-16_4 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-16_4.RAW SUBJECT_SAMPLE_FACTORS 100n-10-16_5 100n-10-16_5 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-16_5.RAW SUBJECT_SAMPLE_FACTORS 100n-10-16_6 100n-10-16_6 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-16_6.RAW SUBJECT_SAMPLE_FACTORS 100n-16-24_1 100n-16-24_1 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-16-24_1.RAW SUBJECT_SAMPLE_FACTORS 100n-16-24_2 100n-16-24_2 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-16-24_2.RAW SUBJECT_SAMPLE_FACTORS 100n-16-24_3 100n-16-24_3 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-16-24_3.RAW SUBJECT_SAMPLE_FACTORS 100n-16-24_4 100n-16-24_4 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-16-24_4.RAW SUBJECT_SAMPLE_FACTORS 100n-16-24_5 100n-16-24_5 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-16-24_5.RAW SUBJECT_SAMPLE_FACTORS 100n-16-24_6 100n-16-24_6 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-16-24_6.RAW SUBJECT_SAMPLE_FACTORS 100n-24-36_1 100n-24-36_1 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-24-36_1.RAW SUBJECT_SAMPLE_FACTORS 100n-24-36_2 100n-24-36_2 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-24-36_2.RAW SUBJECT_SAMPLE_FACTORS 100n-24-36_3 100n-24-36_3 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-24-36_3.RAW SUBJECT_SAMPLE_FACTORS 100n-24-36_4 100n-24-36_4 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-24-36_4.RAW SUBJECT_SAMPLE_FACTORS 100n-24-36_5 100n-24-36_5 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-24-36_5.RAW SUBJECT_SAMPLE_FACTORS 100n-24-36_6 100n-24-36_6 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-24-36_6.RAW SUBJECT_SAMPLE_FACTORS 100n-10-5d_1 100n-10-5d_1 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-5d_1.RAW SUBJECT_SAMPLE_FACTORS 100n-10-5d_2 100n-10-5d_2 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-5d_2.RAW SUBJECT_SAMPLE_FACTORS 100n-10-5d_3 100n-10-5d_3 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-5d_3.RAW SUBJECT_SAMPLE_FACTORS 100n-10-5d_4 100n-10-5d_4 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-5d_4.RAW SUBJECT_SAMPLE_FACTORS 100n-10-5d_5 100n-10-5d_5 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-5d_5.RAW SUBJECT_SAMPLE_FACTORS 100n-10-5d_6 100n-10-5d_6 Treatment:100nM Model=Positive/Negitive; RAW_FILE_NAME=100n-10-5d_6.RAW SUBJECT_SAMPLE_FACTORS 250n-10-16_1 250n-10-16_1 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-16_1.RAW SUBJECT_SAMPLE_FACTORS 250n-10-16_2 250n-10-16_2 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-16_2.RAW SUBJECT_SAMPLE_FACTORS 250n-10-16_3 250n-10-16_3 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-16_3.RAW SUBJECT_SAMPLE_FACTORS 250n-10-16_4 250n-10-16_4 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-16_4.RAW SUBJECT_SAMPLE_FACTORS 250n-10-16_5 250n-10-16_5 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-16_5.RAW SUBJECT_SAMPLE_FACTORS 250n-10-16_6 250n-10-16_6 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-16_6.RAW SUBJECT_SAMPLE_FACTORS 250n-16-24_1 250n-16-24_1 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-16-24_1.RAW SUBJECT_SAMPLE_FACTORS 250n-16-24_2 250n-16-24_2 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-16-24_2.RAW SUBJECT_SAMPLE_FACTORS 250n-16-24_3 250n-16-24_3 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-16-24_3.RAW SUBJECT_SAMPLE_FACTORS 250n-16-24_4 250n-16-24_4 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-16-24_4.RAW SUBJECT_SAMPLE_FACTORS 250n-16-24_5 250n-16-24_5 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-16-24_5.RAW SUBJECT_SAMPLE_FACTORS 250n-16-24_6 250n-16-24_6 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-16-24_6.RAW SUBJECT_SAMPLE_FACTORS 250n-24-36_1 250n-24-36_1 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-24-36_1.RAW SUBJECT_SAMPLE_FACTORS 250n-24-36_2 250n-24-36_2 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-24-36_2.RAW SUBJECT_SAMPLE_FACTORS 250n-24-36_3 250n-24-36_3 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-24-36_3.RAW SUBJECT_SAMPLE_FACTORS 250n-24-36_4 250n-24-36_4 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-24-36_4.RAW SUBJECT_SAMPLE_FACTORS 250n-24-36_5 250n-24-36_5 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-24-36_5.RAW SUBJECT_SAMPLE_FACTORS 250n-24-36_6 250n-24-36_6 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-24-36_6.RAW SUBJECT_SAMPLE_FACTORS 250n-10-5d_1 250n-10-5d_1 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-5d_1.RAW SUBJECT_SAMPLE_FACTORS 250n-10-5d_2 250n-10-5d_2 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-5d_2.RAW SUBJECT_SAMPLE_FACTORS 250n-10-5d_3 250n-10-5d_3 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-5d_3.RAW SUBJECT_SAMPLE_FACTORS 250n-10-5d_4 250n-10-5d_4 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-5d_4.RAW SUBJECT_SAMPLE_FACTORS 250n-10-5d_5 250n-10-5d_5 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-5d_5.RAW SUBJECT_SAMPLE_FACTORS 250n-10-5d_6 250n-10-5d_6 Treatment:250nM Model=Positive/Negitive; RAW_FILE_NAME=250n-10-5d_6.RAW #COLLECTION CO:COLLECTION_SUMMARY Wild-type zebrafish embryos (AB line) were maintained in fish water (0.2 % CO:COLLECTION_SUMMARY Instant Ocean Salt in deionized water, pH 6.5-8.5, conductivity 450-550 μS.cm-1 CO:COLLECTION_SUMMARY and hardness 50-100 mg·L-1 CaCO3) on a 14-h light:10-h dark cycle at 28 °C. CO:COLLECTION_SUMMARY Thirty-three embryos were placed in each well of a 6-well plate (Nest Biotech., CO:COLLECTION_SUMMARY China) in 3 mL fresh fish water. Zebrafish embryos were exposed to DM from CO:COLLECTION_SUMMARY 10-16, 16-24, 24-36 and 10-120 hpf, respectively. DM was washed out after each CO:COLLECTION_SUMMARY restricted time point, and zebrafish were assayed at the larval stage, 120 hpf. CO:COLLECTION_SUMMARY At each exposure stage, zebrafish embryos were given DM dosages of 1, 100, and CO:COLLECTION_SUMMARY 250 nM. The control group received an equivalent volume of dimethyl sulfoxide CO:COLLECTION_SUMMARY (DMSO). In each group, 3 wells of zebrafish were utilized to evaluate CO:COLLECTION_SUMMARY developmental toxicity, locomotor behavior and apoptotic cells in the central CO:COLLECTION_SUMMARY nervous system, while 6 wells of zebrafish were created as separate biological CO:COLLECTION_SUMMARY replicates for subsequent lipidomics analysis and kept immediately at -80 °C. CO:SAMPLE_TYPE Zebrafish #TREATMENT TR:TREATMENT_SUMMARY Wild-type zebrafish embryos (AB line) were maintained in fish water (0.2 % TR:TREATMENT_SUMMARY Instant Ocean Salt in deionized water, pH 6.5-8.5, conductivity 450-550 μS.cm-1 TR:TREATMENT_SUMMARY and hardness 50-100 mg·L-1 CaCO3) on a 14-h light:10-h dark cycle at 28 °C. TR:TREATMENT_SUMMARY Thirty-three embryos were placed in each well of a 6-well plate (Nest Biotech., TR:TREATMENT_SUMMARY China) in 3 mL fresh fish water. Zebrafish embryos were exposed to DM from TR:TREATMENT_SUMMARY 10-16, 16-24, 24-36 and 10-120 hpf, respectively. DM was washed out after each TR:TREATMENT_SUMMARY restricted time point, and zebrafish were assayed at the larval stage, 120 hpf. TR:TREATMENT_SUMMARY At each exposure stage, zebrafish embryos were given DM dosages of 1, 100, and TR:TREATMENT_SUMMARY 250 nM. The control group received an equivalent volume of dimethyl sulfoxide TR:TREATMENT_SUMMARY (DMSO). In each group, 3 wells of zebrafish were utilized to evaluate TR:TREATMENT_SUMMARY developmental toxicity, locomotor behavior and apoptotic cells in the central TR:TREATMENT_SUMMARY nervous system, while 6 wells of zebrafish were created as separate biological TR:TREATMENT_SUMMARY replicates for subsequent lipidomics analysis and kept immediately at -80 °C. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Each replicate of zebrafish (1 well, ~10 mg) was accurately weighted and SP:SAMPLEPREP_SUMMARY transferred to an Eppendorf tube. Then, each sample was spiked with 400 μL of SP:SAMPLEPREP_SUMMARY precooled methanol containing lipid standards (i.e., 1.5 μg/mL of SP:SAMPLEPREP_SUMMARY phosphatidylcholine (PC) (19:0/19:0), 1.5 μg/mL of phosphatidylethanolamine SP:SAMPLEPREP_SUMMARY (PE) (15:0/15:0), 1.5 μg/mL of lysophospatidylcholine (LPC) (19:0), 1.5 μg/mL SP:SAMPLEPREP_SUMMARY of sphingomyelin (SM) (d18:1/12:0), 1.2 μg/mL of triacylglycerol (TG) SP:SAMPLEPREP_SUMMARY (15:0/15:0/15:0), 1.2 μg/mL of ceramide (Cer) (d18:1/17:0), 1 μg/mL of SP:SAMPLEPREP_SUMMARY palmitic acid (fatty acid (FA) (16:0))-d3, and 1 μg/mL of stearic acid SP:SAMPLEPREP_SUMMARY (FA(18:0))-d3), followed by tissue homogenization using the bead-based SP:SAMPLEPREP_SUMMARY homogenizer (Tissuelyser-24, Shanghai jingxin industrial development co., ltd, SP:SAMPLEPREP_SUMMARY China) for 1 min at 65 Hz. Next, 1 mL of tert-butyl methyl ether (MTBE) was SP:SAMPLEPREP_SUMMARY added to the tube, and the mixture was thoroughly vortexed (1,000 rpm, 10 ℃, SP:SAMPLEPREP_SUMMARY 30 min). Phase separation was induced by adding 400 μL of Milli-Q water and SP:SAMPLEPREP_SUMMARY centrifugation at 15,000 g for 15 min at 6 ℃. The upper hydrophobic phase was SP:SAMPLEPREP_SUMMARY collected and vacuum-dried in in a refrigerated CentriVap concentrator SP:SAMPLEPREP_SUMMARY (Labconco, USA), followed by storage at -80 °C until subsequent analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) CH:SOLVENT_A Acetonitrile/water (6:4, v/v) CH:SOLVENT_B Isopropanol/acetonitrile (9:1, v/v) CH:FLOW_GRADIENT First maintain 32% B for 1.5 minutes, then linearly increase from 1.5 to 15.5 CH:FLOW_GRADIENT minutes to 85% B, then linearly increase to 97% B at 0.1 minutes, maintain for CH:FLOW_GRADIENT 18 minutes, and then rapidly decrease to 32% B at 0.1 minutes, equilibrate until CH:FLOW_GRADIENT the next injection CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 55 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Exploris120 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Xcalibular MS:MS_RESULTS_FILE ST002981_AN004900_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END