Summary of Study ST000598

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000436. The data can be accessed directly via it's Project DOI: 10.21228/M8PW36 This work is supported by NIH grant, U2C- DK119886.

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Study IDST000598
Study TitleDysfunctional lipid metabolism underlies the effect of the perinatal DDT exposure on the development of metabolic syndrome (part II)
Study SummaryThis study aims to identify changes in lipid mediators in the hypothalamus with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP (n=8 per group) or not treated (n=8 per group). Each group was analyzed for oxylipin, nitro lipids, endocannabinoid, and endocannabinoid-like monoacylglycerol and N-acylethanolamide changes to investigate alterations in lipid mediator signaling due to TPP exposure. Targeted metabolomic analysis of lipid mediators in rat hypothalamus samples was performed by the Newman lab.
Institute
University of California, Davis
DepartmentUSDA Western Human Nutrition Research Center
LaboratoryNewman Lab
Last NameNewman
First NameJohn
Address430 W. Health Sciences Dr., Davis, CA 95616
Emailjohn.newman@ars.usda.gov
Phone+1-530-752-1009
Submit Date2017-04-19
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2017-07-10
Release Version1
John Newman John Newman
https://dx.doi.org/10.21228/M8PW36
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000436
Project DOI:doi: 10.21228/M8PW36
Project Title:Dysfunctional lipid metabolism underlies the effect of the perinatal DDT exposure on the development of metabolic syndrome
Project Summary:This study aims to identify changes in lipid mediators in the hypothalamus with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP (n=8 per group) or not treated (n=8 per group). Each group was analyzed for oxylipin, nitro lipids, endocannabinoid, and endocannabinoid-like monoacylglycerol and N-acylethanolamide changes to investigate alterations in lipid mediator signaling due to TPP exposure. Targeted metabolomic analysis of lipid mediators in rat hypothalamus samples was performed by the Newman lab.
Institute:University of California, Davis
Department:Department of Environmental Toxicology
Last Name:La Merrill
First Name:Michele
Address:1 Shields Ave., Davis, CA 95616
Email:mlamerrill@ucdavis.edu
Phone:(530) 754-7254

Subject:

Subject ID:SU000621
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammal

Factors:

Subject type: Animal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
SA032766LAM-03E (vehicle control)
SA032767LAM-16E (vehicle control)
SA032768LAM-14E (vehicle control)
SA032769LAM-15E (vehicle control)
SA032770LAM-01&09 avg.E (vehicle control)
SA032771LAM-07E (vehicle control)
SA032772LAM-08E (vehicle control)
SA032773LAM-13E (vehicle control)
SA032774LAM-17T (treated with TPhP)
SA032775LAM-02T (treated with TPhP)
SA032776LAM-06T (treated with TPhP)
SA032777LAM-11&12 avg.T (treated with TPhP)
SA032778LAM-10T (treated with TPhP)
SA032779LAM-18T (treated with TPhP)
SA032780LAM-04T (treated with TPhP)
SA032781LAM-05T (treated with TPhP)
Showing results 1 to 16 of 16

Collection:

Collection ID:CO000615
Collection Summary:"Prior to sacrifice, rats were fasted between 8 and12 h,theirbody weights recorded, and blood was collected from the tail as above prior to euthanasia. Rats were anesthetized with sodium pentobarbital and euthanized with a 1 mL/kg intracardiac injection of saturated potassium chloride. Once cardiac movement had stopped for 30 s the rat was decapitated and the hypothalamus, liver, pancreas, heart, mesenteric adipose tissue, quadriceps, kidney, gonadal adipose tissue, inguinal adipose tissue, and brown adipose tissue were collected. All tissues were removed in the order listed above, wet weighed, and snap frozen in liquid nitrogen"
Collection Protocol Filename:Green_et_al_2017-TPP_Exposure_Accelerat_T2DM_Rats.pdf
Sample Type:Tissue

Treatment:

Treatment ID:TR000635
Treatment Summary:"Adult non-pregnant female UCD-T2DM rats (n = 16; 3 months old) were paired with males (n = 10; 3–4 months old) for a 24 h period at which point males were removed. This was defined as gestational day zero (G0) if a sperm plug was observed or if the female rats gained at least 30 g of body weight over the next 7 days. The day of birth was designated postnatal day zero (P0). Pregnant dams were randomly assigned to an exposure group (n = 8 per group), and received daily oral TPhP or ethanol vehicle exposure from G8 through weaning (P21) as described in Section 2.2 below. Gestational length and litter size were recorded on P0 and the sex of pups was determined and recorded on P4. Body weights of all pups in each litter were obtained periodically from P4–21. On P4 the litters were culled to 8 pups ensuring up to 4males and 2 females in each litter by random selection (Fig. 1A & B). This was done to ensure consistent exposure of pups between litters [13,23]. The time ittakes to develop T2DM is accelerated among UCD-T2DM rats with higher body weights on P21. Hence at weaning the largest pups were housed in same sex littermate groups of two females and up to four males as available (Fig. 1A & B). Urine was collected from the dams using an adapted plastic wrap method outlined by Kurien [24], 60 mins after final dose. Dams were placed in clean cages without bedding for at least 20 min then using a pipette up to 500 L of urine was collected in ethanol rinsed glass vials and placed on ice. At weaning all dams and remaining weanlings were sacrificed (90–330 min post-exposure) by CO2 asphyxiation and rapid decapitation. Two male rats weighing between 350–400 g on P61, from the TPhP group and the vehicle group were weight-matched across treatments for the diabetes study to eliminate confounding effects of body mass on the association between TPhP and T2DM onset (Fig. 1B). This weight range was selected because male UCD-T2DM rats that are between 350 and 400 g at 8 weeks of age develop T2DM at approximately 23 weeks of age [18]. Weight-matched rats were followed until 26 weeks or until they developed T2DM, which was defined as two consecutive weekly non-fasting glucose measurements of ≥200 mg/dL [18] in accordance with the American Diabetes Association (ADA) guideline of diagnosing diabetes with a random plasma glucose of 200 mg/dL or higher [19]. The remaining rats were not weight-matched and followed for the 3.5 month obesity study (Fig. 1A).
Treatment Protocol Filename:Green_et_al_2017-TPP_Exposure_Accelerat_T2DM_Rats.pdf

Sample Preparation:

Sampleprep ID:SP000628
Sampleprep Summary:Oxylipins and endocannabinoids were isolated using a Waters Ostro Sample Preparation Plate (Milford, MA). Hypothalamus samples were pulverized and aliquoted (~20-25mg) were added to 2mL polypropylene tubes and spiked with a 5 µL anti-oxidant solution (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water) and 10 μL 1000nM analytical deuterated surrogates. A total of 100 µL methanol was added to the sample and vrtexed 90 sec. Next, 500 µL D.I. water and 1000 µL ethyl acetate was added and the tube was vortexed 3 minutes, before being centrifuged at 15,000g for 10 min at room temp. The supernate was then transferred into a clean 2 mL autosampler vial. The extraction with ethyl acetate was repeated and the eluent was dried by speed vacuum for 35 min at the medium BP setting. Once dry, samples were re-constituted with the internal standard 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU) at 100 nM (50:50 MeOH:CAN), vortexed 1 min, transferred to a spin filter (0.1 µm, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf), before being transferred to 2 mL LC-MS amber vials. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of surrogate standards.
Sampleprep Protocol Filename:Hypothal_Newman_Data_Report.docx

Combined analysis:

Analysis ID AN000916
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex API 4000 QTrap
Ion Mode NEGATIVE
Units Concentration pmol/g

Chromatography:

Chromatography ID:CH000653
Instrument Name:Waters Acquity
Column Name:Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)
Chromatography Type:Reversed phase

MS:

MS ID:MS000815
Analysis ID:AN000916
Instrument Name:ABI Sciex API 4000 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
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