Summary of Study ST002304
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 PR001477. The data can be accessed directly via it's Project DOI: 10.21228/M8T40Q This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
Study ID | ST002304 |
Study Title | White-nose syndrome disrupts the splenic lipidome of little brown bats (Myotis lucifugus) at early disease stages |
Study Summary | The fungal disease of bats, white-nose syndrome (WNS), is caused by the pathogen Pseudogymnoascus destructans (Pd). WNS-positive little brown bats (Myotis lucifugus) can exhibit an immune response during infection that include increases in cytokine and pro-inflammatory mediator gene levels. While bioactive lipid mediators (oxylipins) formed by enzymatic oxidation of polyunsaturated fatty acids (PUFAs) can contribute to this type of immune response, their role in WNS pathophysiology have not been investigated. Nonenzymatic conversion of PUFAs can also occur due to reactive oxygen species (ROS), however, these enantiomeric isomers will lack the same signaling properties. In this study, we performed a series of targeted lipidomic approaches on laboratory Pd-inoculated bats to assess changes in their splenic lipidome, including the formation of lipid mediators at early stages of WNS. Hepatic lipids previously identified were also resolved to a higher structural detail. We compared WNS-susceptible M. lucifugus to a WNS-resistant species, the big brown bat (Eptesicus fuscus). Altered splenic lipid levels were only observed in M. lucifugus, with lower total levels of glycerophospholipids (GPs) and free fatty acids (FFAs) in the Pd-inoculated group compared to the sham-inoculated group. Lower concentrations of splenic GPs were observed in lipid compounds containing 18:2 or saturated acyl chains. Differences in splenic FFAs included both omega-3 (including docosahexaenoic acid [DHA]) and omega-6 compounds. Increased levels of an enantiomeric monohydroxy DHA (4-hydroxydocosahexaenoic [HDoHE], 10-HDoHE, and 13-HDoHE) mixture, suggesting nonenzymatic formation, along with 6-keto-PGF1a were found. Changes in previously identified hepatic lipids were confined to omega-3 constituents. Together, these results suggest that increased oxidative stress, but not an inflammatory response, is occurring in bats at early stages of WNS that proceeds fat depletion. |
Institute | Georgetown University |
Last Name | Pannkuk |
First Name | Evan |
Address | 3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA |
elp44@georgetown.edu | |
Phone | 2026875650 |
Submit Date | 2022-09-22 |
Analysis Type Detail | LC-MS |
Release Date | 2023-09-22 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP002396 |
Sampleprep Summary: | Tissue samples were homogenized in 300 μL of extraction buffer (isopropanol [IPA]) containing internal standards for the above lipid classes per the manufacturer’s instructions. A 10 μL aliquot of the tissue/extraction buffer mixture was collected for determining protein concentration. The samples were vortexed for 30 seconds, incubated on ice for 30 min, and then centrifuged for 10 min (10,000 x g, 4°C). The supernatant was transferred to a MS vial for LC-MS analysis. A pooled sample of all aliquots was prepared as a quality control and run every 10 samples in addition to the NIST SRM 1950 plasma mix. The NIST SRM 1950 plasma was prepared by aliquoting 20 µL in 100 μL of chilled IPA containing internal standards. Samples were vortexed for 1 min, incubated on ice for 30 min then incubated at -20°C for 2 hours for complete protein precipitation, then centrifuged for 20 min (10,000 x g, 4°C). |