Summary of Study ST001106
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 PR000740. The data can be accessed directly via it's Project DOI: 10.21228/M81D57 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST001106 |
| Study Title | Lipidomics of Newborn Heart Tissue Exposed to Excess Maternal Cortisol in Late Gestation (part-1) |
| Study Summary | Cardiac tissue from newborn hearts from animals exposed to excess maternal cortisol in late gestation and untreated was compared via MS lipidomic analysis |
| Institute | University of Florida |
| Department | Biochemsitry & Molecular Biology |
| Last Name | Walejko |
| First Name | Jacquelyn |
| Address | R3-226 Academic Research Building, Department of Biochemistry and Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245 |
| jwalejko@ufl.edu | |
| Phone | na |
| Submit Date | 2018-11-30 |
| Num Groups | 2 |
| Total Subjects | 12 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | fid |
| Analysis Type Detail | LC-MS |
| Release Date | 2019-03-06 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000740 |
| Project DOI: | doi: 10.21228/M81D57 |
| Project Title: | Chronic Maternal Cortisol Excess During Late Gestation Leads to Metabolic Alterations in the Newborn Heart |
| Project Summary: | Our laboratory has previously shown in an ovine model of pregnancy that abnormal elevations in maternal cortisol during late gestation lead to increased fetal cardiac arrhythmias and mortality during peripartum. Furthermore, transcriptomic analysis of the fetal heart suggested alterations in TCA cycle intermediates and lipid metabolites in animals exposed to excess cortisol in utero. Therefore, we utilized a sheep model of pregnancy to determine how chronic increases in maternal cortisol alter maternal and fetal serum prior to birth and neonatal cardiac metabolites and lipids at term. Ewes were either infused with 1 mg/kg/day of cortisol starting at gestational day 115 (n=9), or untreated (n=6). Serum was collected from the mother and fetus (125 d-birth), and hearts were collected following birth. Proton nuclear magnetic resonance (1H-NMR) spectroscopy was conducted to measure metabolic profiles of newborn heart specimens as well as fetal and maternal serum specimens. Mass spectrometry was conducted to measure lipid profiles of newborn heart specimens. We observed alterations in amino acid and TCA cycle metabolism as well as lipid and glycerophospholipid metabolism in newborn hearts after excess maternal cortisol in late gestation. In addition, we observed alterations in amino acid and TCA cycle metabolites in fetal but not in maternal serum during late gestation. These results suggest that fetal exposure to excess maternal cortisol alters placental and fetal metabolism prior to birth and limits normal cardiac metabolic maturation, which may contribute to increased risk of peripartum cardiac arrhythmias observed in these animals, or later life cardiomyopathies. |
| Institute: | University of Florida |
| Department: | Pharmacodynamics |
| Last Name: | Keller-Wood |
| First Name: | Maureen |
| Address: | 1345 SW Archer Rd, PO 100487, Gainesville, FL, 32610 |
| Email: | kellerwd@cop.ufl.edu |
| Phone: | NA |
Subject:
| Subject ID: | SU001151 |
| Subject Type: | Mammal |
| Subject Species: | Ovis aries |
| Taxonomy ID: | 9940 |
| Age Or Age Range: | within 6 hours of birth |
Factors:
Subject type: Mammal; Subject species: Ovis aries (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA075241 | 767 | Control |
| SA075242 | 1577 | Control |
| SA075243 | 1610-16 | Control |
| SA075244 | 753 | Control |
| SA075245 | 770 | Control |
| SA075246 | 734 | Control |
| SA075247 | 1576 | Cortisol |
| SA075248 | 3843 | Cortisol |
| SA075249 | 717 | Cortisol |
| SA075250 | 729 | Cortisol |
| SA075251 | 706 | Cortisol |
| SA075252 | 802 | Cortisol |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO001145 |
| Collection Summary: | Immediately following birth, the lambs were euthanized with an overdose of Euthasol (pentobarbital sodium and phenytoin sodium; Virbac AH Inc), and heart tissue was collected from the right ventricle (RV), left ventricle (LV), and intraventricular septum (IVS) and immediately frozen in liquid nitrogen. Tissue samples were stored at -80 °C until data collection. |
| Sample Type: | Cardiac tissue |
| Collection Method: | Heart tissue was collected under sterile conditions and immediately frozen in liquid nitrogen |
| Collection Location: | University of Florida |
| Storage Conditions: | -80℃ |
| Collection Vials: | Cryovials |
| Storage Vials: | Cryovials |
Treatment:
| Treatment ID: | TR001165 |
| Treatment Summary: | The treatment protocol for this study was previous published in Antolic, A., et al. (2018). Chronic maternal hypercortisolemia in late gestation alters fetal cardiac function at birth. Am J Physiol Regul Integr Comp Physiol 314(3): R342-R352. |
| Treatment Compound: | Hydrocortisone sodium succinate in sodium phosphate (Solu-Cortef; Pfizer, New York, NY, USA) |
| Treatment Dose: | 1 mg/kg/day |
Sample Preparation:
| Sampleprep ID: | SP001158 |
| Sampleprep Summary: | Homogenized right ventricle tissue samples (100 mg from 7 fetuses and 7 newborns) were used for lipid extraction via the Folch method. Briefly, 2 mL of internal standard mixture was added to each sample before adding 1.2 mL of 2:1 chloroform/methanol solution (HPLC-grade). Samples were incubated at 4 °C for 20 min with occasional vortexing. Following incubation, 200 mL of water (HPLC-grade) was added and samples were incubated at 4 °C for 10 min. Samples were centrifuged at 2,000 g for 5 min at 4 °C, and the resulting chloroform layer was removed and transferred into a clean tube. The extraction process was repeated with 400 mL 2:1 chloroform/methanol (HPLC-grade) and the resulting chloroform layers were combined. Samples were dried under a nitrogen stream at 30 °C and stored at -80 °C until reconstitution. Samples were reconstituted with 200 mL of isopropanol and 2 mL injection standard mixture. Ammonium acetate and all analytical grade solvents (formic acid, chloroform, and methanol) were purchased from Fisher Scientific (Waltham, MA, USA). All mobile phase solvents were Fisher Optima LC/MS-grade (acetonitrile, isopropanol, and water). Organ: Heart, Organ Specification: Right Ventricle |
| Processing Method: | 50 mg of tissue was weighed and added to tubes with three 3mm glass beads, two 5mm steal beads, and 0.7mm zirconia beads (“2 squirts”). HPLC-water (1mL) was added to each tube before homogenization at 1800 rpm for 30 seconds for 5 rounds. Samples were incubated at 4C for 20 min between rounds. Following homogenization, samples were centrifuged at 2,000g for 10 min at 4C to pellet tissue debris before transferring supernatant to a clean tube. Homogenates were stored at -80C. |
| Extraction Method: | 2 mL of internal standard mixture was added to each sample before adding 1.2 mL of 2:1 chloroform/methanol solution (HPLC-grade). Samples were incubated at 4 °C for 20 min with occasional vortexing. Following incubation, 200 mL of water (HPLC-grade) was added and samples were incubated at 4 °C for 10 min. Samples were centrifuged at 2,000 g for 5 min at 4 °C, and the resulting chloroform layer was removed and transferred into a clean tube. The extraction process was repeated with 400 mL 2:1 chloroform/methanol (HPLC-grade) and the resulting chloroform layers were combined. Samples were dried under a nitrogen stream at 30 °C (Organomation Associated MultiVap) and stored at -80 °C until reconstitution. |
| Sample Resuspension: | Samples were reconstituted with 200 mL of isopropanol and 2 mL injection standard mixture and vortexed to mix. Fifty uL of reconstituted sample was transferred to a glass LC vial for analysis. |
| Sample Spiking: | Internal standards: lysophosphatidylcholine (17:0), phosphatidylcholine (17:0/17:0), phosphatidylglycerol (14:0/14:0), phosphatidylethanolamine (15:0/15:0), phosphatidylserine (14:0/14:0), triglyceride (15:0/15:0/15:0). Injection Standards: lysophosphatidylcholine (19:0), phosphatidylcholine (19:0/19:0), phosphatidylglycerol (17:0/17:0), phosphatidylethanolamine (17:0/17:0), phosphatidylserine (17:0/17:0), triglyceride (17:0/17:0/17:0). |
Combined analysis:
| Analysis ID | AN001799 | AN001800 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Relative peak area | Relative peak area |
Chromatography:
| Chromatography ID: | CH001270 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) |
| Column Temperature: | 4C |
| Flow Gradient: | 80% A and 20% B at 0 min; 80% A and 20% B at 1 min; 70% A and 30% B at 3 min; 55% A and 45% B at 4 min; 40% A and 60% B at 6 min; 35% A and 65% B at 8 min; 35% A and 65% B at 10 min; 10% A and 90% B at 15 min; 2% A and 98% B at 17 min; 2% A and 98% B at 18 min; 80% A and 20% B at 19 min; 80% A and 20% B at 23 min |
| Flow Rate: | 0.5mL/min |
| Sample Injection: | 2 uL |
| Solvent A: | 40% water/60% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/8% acetonitrile/2% water; 0.1% formic acid; 10 mM ammonium formate |
| Analytical Time: | 18 min |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001660 |
| Analysis ID: | AN001799 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 250C |
| Ion Spray Voltage: | 1500 |
| Ionization: | Heated electrospray |
| Resolution Setting: | 70,000 |
| Scanning Range: | 200-2200 m/z |
| MS ID: | MS001661 |
| Analysis ID: | AN001800 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 250C |
| Ion Spray Voltage: | 1500 |
| Ionization: | Heated electrospray |
| Resolution Setting: | 70,000 |
| Scanning Range: | 200-2200 m/z |
