Summary of Study ST001245

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 PR000832. The data can be accessed directly via it's Project DOI: 10.21228/M84Q24 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001245
Study TitleLuteal lipids regulate progesterone production and may modulate immune cell function during the estrous cycle and pregnancy
Study SummaryDespite data indicating an important functional role for bioactive lipids in luteal function, little is known about the patterns of abundance of these lipids in corpus luteum (CL) during luteal development, maintenance, and rescue, in any species. Therefore, the abundance of lipid mediators, including endocannabinoids and oxylipins from cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P450 (CYP)-dependent metabolism were profiled in the CL on days 4, 11, and 18 of the estrous cycle and on day 18 of pregnancy. The objectives of this study were to identify lipid mediators that regulate luteal function during these transitions, to integrate the lipid profile with a previously published mRNA profile of CL during maternal recognition of pregnancy, and to determine the effect of a subset of lipids on in vitro progesterone production.
Institute
University of California, Davis
DepartmentGenome and Biomedical Sciences Facility
LaboratoryWCMC Metabolomics Core
Last NameFiehn
First NameOliver
Address1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Emailofiehn@ucdavis.edu
Phone(530) 754-8258
Submit Date2019-07-29
PublicationsHughes, C.H.K., R. Bosviel, J.W. Newman, J.L. Pate. Luteal lipids regulate progesterone production and may modulate immune cell function during the estrous cycle and pregnancy. Front. Endocrinol. Jun 27, 2019
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2019-09-10
Release Version1
Oliver Fiehn Oliver Fiehn
https://dx.doi.org/10.21228/M84Q24
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000832
Project DOI:doi: 10.21228/M84Q24
Project Title:Luteal lipids regulate progesterone production and may modulate immune cell function during the estrous cycle and pregnancy
Project Summary:Despite data indicating an important functional role for bioactive lipids in luteal function, little is known about the patterns of abundance of these lipids in corpus luteum (CL) during luteal development, maintenance, and rescue, in any species. Therefore, the abundance of lipid mediators, including endocannabinoids and oxylipins from cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P450 (CYP)-dependent metabolism were profiled in the CL on days 4, 11, and 18 of the estrous cycle and on day 18 of pregnancy. The objectives of this study were to identify lipid mediators that regulate luteal function during these transitions, to integrate the lipid profile with a previously published mRNA profile of CL during maternal recognition of pregnancy, and to determine the effect of a subset of lipids on in vitro progesterone production.
Institute:University of California, Davis
Department:Genome and Biomedical Sciences Facility
Laboratory:WCMC Metabolomics Core
Last Name:Fiehn
First Name:Oliver
Address:1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616
Email:ofiehn@ucdavis.edu
Phone:(530) 754-8258
Funding Source:NIH U24DK097154

Subject:

Subject ID:SU001313
Subject Type:Mammal
Subject Species:Bos taurus
Taxonomy ID:9913
Gender:Female

Factors:

Subject type: Mammal; Subject species: Bos taurus (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
SA090787971D18 cyclic
SA0907881022D18 cyclic
SA0907891008D18 cyclic
SA0907901012D18 cyclic
SA0907911011D18 pregnant
SA0907921021D18 pregnant
SA0907931020D18 pregnant
SA0907941025D18 pregnant
SA0907951005Early
SA0907961106Early
SA0907971006Early
SA0907981007Early
SA090799957Midcycle
SA090800936Midcycle
SA0908011129Midcycle
SA090802956Midcycle
SA090806876Regressing 12 hour
SA090807912Regressing 12 hour
SA090808909Regressing 12 hour
SA090809905Regressing 12 hour
SA090803604Regressing 1 hour
SA090804600Regressing 1 hour
SA090805858Regressing 1 hour
SA0908101379Regressing 24 hour
SA0908111378Regressing 24 hour
SA0908121380Regressing 24 hour
SA0908131381Regressing 24 hour
SA090814616Regressing 4 hour
SA090815575Regressing 4 hour
SA090816862Regressing 4 hour
SA090817857Regressing 4 hour
SA0908181352Regressing 8 hour
SA0908191342Regressing 8 hour
SA0908201351Regressing 8 hour
SA0908211353Regressing 8 hour
Showing results 1 to 35 of 35

Collection:

Collection ID:CO001307
Collection Summary:For cows assigned to the day 4 group, upon observation of estrus and a dominant follicle by ultrasound, cows were given an injection of GnRH (Factrel, 100 µg; Zoetis) in order to precisely time ovulation relative to time of collection for these early CL. Cows were slaughtered on day 4 following estrus. For samples collected later than day 4, precise synchrony of ovulation relative to CL collection was not necessary, so no GnRH was given, and CL were collected via colpotomy. For CL of pregnancy, cows were bred by artificial insemination and a uterine flush was performed immediately following CL collection and was examined for embryo fragments to confirm the presence of a viable pregnancy. For all samples, tissue was snap frozen in liquid nitrogen immediately following tissue collection and stored at -80 degrees Celsius thereafter. For in vitro experiments, three to five dairy cows were used in each group, CL were collected on day 10-12 of the estrous cycle, and each treatment was applied to cells from each cow
Sample Type:Corpus Luteum

Treatment:

Treatment ID:TR001328
Treatment Summary:For cows assigned to the day 4 group, upon observation of estrus and a dominant follicle by ultrasound, cows were given an injection of GnRH (Factrel, 100 µg; Zoetis) in order to precisely time ovulation relative to time of collection for these early CL. Cows were slaughtered on day 4 following estrus. For samples collected later than day 4, precise synchrony of ovulation relative to CL collection was not necessary, so no GnRH was given, and CL were collected via colpotomy. For CL of pregnancy, cows were bred by artificial insemination and a uterine flush was performed immediately following CL collection and was examined for embryo fragments to confirm the presence of a viable pregnancy. For all samples, tissue was snap frozen in liquid nitrogen immediately following tissue collection and stored at -80 degrees Celsius thereafter. For in vitro experiments, three to five dairy cows were used in each group, CL were collected on day 10-12 of the estrous cycle, and each treatment was applied to cells from each cow

Sample Preparation:

Sampleprep ID:SP001321
Sampleprep Summary:Oxylipins and endocannabinoids were isolated using a Waters Ostro™ Sample Preparation Plate. Luteal samples were homogenized and 40 ± 8 mg were added to 2 mL polypropylene tubes spiked with a 5 µL antioxidant solution (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water) and 10 μL 1000 nM analytical deuterated surrogates as previously described (Agrawal et al., 2017; La Frano et al., 2017). Samples were then mixed with 35 µL methanol, 550 µL isopropanol w/ 10 mM ammonium formate, 1% formic acid and 100 µL water, and the tube was placed in a Geno/Grinder 2010 (SPEX SamplePrep) for 30 sec and centrifuged at 10,000 x g for 5 min at room temperature. Supernatants were transferred into the Ostro plate wells and captured in glass inserts containing 10 μL of 20% glycerol in methanol by applying 15 mmHg of vacuum for 10 min. The eluent was dried under vacuum and reconstituted with 100 µL, 1:1 MeOH/ACN (v/v) containing 100 nM of 1-cyclohexyl ureido, 3 dodecanoic acid and 1-phenyl ureido, 3-hexanoic acid urea used as internal standards (gifts from Dr. B.D. Hammock, University of California, Davis). The samples were then vortexed and filtered at 0.1µm through PVDF membranes (Millipore) by centrifugation < 4500 x g (rcf) for 3 min at 6 ºC. The filtrate was transferred to inserts in amber glass and stored at -20 ºC for less than 48 hours before analysis by UPLC-MS/MS. Analytes in 5 μL extract aliquot were separated on a 2.1 mm x 150 mm, 1.7 µm Acquity BEH column (Waters) using published protocols for oxylipins and endocannabinoids (Agrawal et al., 2017; Pedersen and Newman, 2018). Samples were held at 10ºC. Separated residues were detected by negative mode electrospray ionization for oxylipins and positive mode electrospray ionization for endocannabinoids using multiple reaction monitoring on an API 6500 QTRAP (AB Sciex). Analytes were quantified using internal standard methods and 5- to 7-point calibration curves (r2 ≥ 0.997). Calibrants and internal standards were either synthesized [10,11-DHN, 10,11-DHHep, 10(11)-EpHep] or purchased from Cayman Chemical, Avanti Polar Lipids Inc., or Larodan Fine Lipids. Data was processed with AB Sciex MultiQuant version 3.0.2. The internal standards were used to quantify recovery of surrogate standards

Combined analysis:

Analysis ID AN002068 AN002069
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Shimadzu Nexera X2 Shimadzu Nexera X2
Column Waters Acquity BEH C18 (150 x 2mm,1.7um) Waters Acquity BEH C18 (150 x 2mm,1.7um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name ABI Sciex 6500 QTrap ABI Sciex 6500 QTrap
Ion Mode POSITIVE NEGATIVE
Units nM nM

Chromatography:

Chromatography ID:CH001506
Instrument Name:Shimadzu Nexera X2
Column Name:Waters Acquity BEH C18 (150 x 2mm,1.7um)
Column Temperature:60
Flow Rate:0.5 mL/min
Chromatography Type:Reversed phase

MS:

MS ID:MS001919
Analysis ID:AN002068
Instrument Name:ABI Sciex 6500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Multiquant
Ion Mode:POSITIVE
  
MS ID:MS001920
Analysis ID:AN002069
Instrument Name:ABI Sciex 6500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Multiquant
Ion Mode:NEGATIVE
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