Summary of Study ST003437

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 PR002120. The data can be accessed directly via it's Project DOI: 10.21228/M8KN7C This work is supported by NIH grant, U2C- DK119886.

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Study IDST003437
Study TitleWhite adipose tissue remodeling in Little Brown Myotis (Myotis lucifugus) with white-nose syndrome
Study SummaryWhite-nose syndrome (WNS) is a fungal wildlife disease of bats that has caused precipitous declines in certain Nearctic bat species. A key driver of mortality is premature exhaustion of fat reserves, primarily white adipose tissue (WAT), that bats rely on to meet their metabolic needs during winter. However, the pathophysiological and metabolic effects of WNS have remained ill-defined. To elucidate metabolic mechanisms associated with WNS mortality, we infected a WNS susceptible species, the Little Brown Myotis (Myotis lucifugus), with Pseudogymnoascus destructans (Pd) and collected WAT biopsies for histology and targeted lipidomics. These results were compared to the WNS-resistant Big Brown Bat (Eptesicus fuscus). A similar distribution in broad lipid class was observed in both species, with total WAT primarily consisting of triacylglycerides. Baseline differences in WAT chemical composition between species showed that higher glycerophospholipids (GPs) levels in E. fuscus were dominated by unsaturated or monounsaturated moieties and n-6 (18:2, 20:2, 20:3, 20:4) fatty acids. Conversely, higher GP levels in M. lucifugus WAT were primarily compounds containing n-3 (20:5 and 22:5) fatty acids. Following Pd-infection, we found that perturbation to WAT reserves occurs in M. lucifugus, but not in the resistant E. fuscus. A total of 66 GPs (primarily glycerophosphocholines and glycerophosphoethanolamines) were higher in Pd-infected M. lucifugus, indicating perturbation to the WAT structural component. In addition to changes in lipid chemistry, smaller adipocyte sizes and increased extracellular matrix deposition was observed in Pd-infected M. lucifugus. This is the first study to describe WAT GP composition of bats with different susceptibilities to WNS and highlights that recovery from WNS may require repair from adipose remodeling in addition to replenishing depot fat during spring emergence.
Institute
Georgetown University
Last NamePannkuk
First NameEvan
Address3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Emailelp44@georgetown.edu
Phone2026875650
Submit Date2024-08-20
Analysis Type DetailLC-MS
Release Date2025-01-02
Release Version1
Evan Pannkuk Evan Pannkuk
https://dx.doi.org/10.21228/M8KN7C
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002120
Project DOI:doi: 10.21228/M8KN7C
Project Title:White adipose tissue remodeling in Little Brown Myotis (Myotis lucifugus) with white-nose syndrome
Project Summary:White-nose syndrome (WNS) is a fungal wildlife disease of bats that has caused precipitous declines in certain Nearctic bat species. A key driver of mortality is premature exhaustion of fat reserves, primarily white adipose tissue (WAT), that bats rely on to meet their metabolic needs during winter. However, the pathophysiological and metabolic effects of WNS have remained ill-defined. To elucidate metabolic mechanisms associated with WNS mortality, we infected a WNS susceptible species, the Little Brown Myotis (Myotis lucifugus), with Pseudogymnoascus destructans (Pd) and collected WAT biopsies for histology and targeted lipidomics. These results were compared to the WNS-resistant Big Brown Bat (Eptesicus fuscus). A similar distribution in broad lipid class was observed in both species, with total WAT primarily consisting of triacylglycerides. Baseline differences in WAT chemical composition between species showed that higher glycerophospholipids (GPs) levels in E. fuscus were dominated by unsaturated or monounsaturated moieties and n-6 (18:2, 20:2, 20:3, 20:4) fatty acids. Conversely, higher GP levels in M. lucifugus WAT were primarily compounds containing n-3 (20:5 and 22:5) fatty acids. Following Pd-infection, we found that perturbation to WAT reserves occurs in M. lucifugus, but not in the resistant E. fuscus. A total of 66 GPs (primarily glycerophosphocholines and glycerophosphoethanolamines) were higher in Pd-infected M. lucifugus, indicating perturbation to the WAT structural component. In addition to changes in lipid chemistry, smaller adipocyte sizes and increased extracellular matrix deposition was observed in Pd-infected M. lucifugus. This is the first study to describe WAT GP composition of bats with different susceptibilities to WNS and highlights that recovery from WNS may require repair from adipose remodeling in addition to replenishing depot fat during spring emergence.
Institute:Georgetown University
Last Name:Pannkuk
First Name:Evan
Address:3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Email:elp44@georgetown.edu
Phone:2026875650

Subject:

Subject ID:SU003564
Subject Type:Mammal
Subject Species:Myotis lucifugus
Taxonomy ID:59463
Species Group:Mammals

Factors:

Subject type: Mammal; Subject species: Myotis lucifugus (Factor headings shown in green)

mb_sample_id local_sample_id Treatment Sample source
SA3783021_S_018PBST white adipose tissue
SA3783031_S_006PBST white adipose tissue
SA3783041_S_024PBST white adipose tissue
SA3783051_S_023PBST white adipose tissue
SA3783061_S_020PBST white adipose tissue
SA3783071_S_019PBST white adipose tissue
SA3783081_S_001PBST white adipose tissue
SA3783091_S_015PBST white adipose tissue
SA3783101_S_007PBST white adipose tissue
SA3783111_S_013PBST white adipose tissue
SA3783121_S_025PBST white adipose tissue
SA3783131_S_008PBST white adipose tissue
SA3783141_S_010PBST white adipose tissue
SA3783151_S_011PBST white adipose tissue
SA3783162_S_002Pd white adipose tissue
SA3783172_S_003Pd white adipose tissue
SA3783182_S_004Pd white adipose tissue
SA3783192_S_009Pd white adipose tissue
SA3783202_S_012Pd white adipose tissue
SA3783212_S_014Pd white adipose tissue
SA3783222_S_016Pd white adipose tissue
SA3783232_S_017Pd white adipose tissue
SA3783242_S_021Pd white adipose tissue
SA3783252_S_022Pd white adipose tissue
SA3783262_S_005Pd white adipose tissue
Showing results 1 to 25 of 25

Collection:

Collection ID:CO003557
Collection Summary:The WAT samples used in this study were collected as part of a captive hibernation experiment performed at Bucknell University from November 2011 to March 2013. The bats were captured between 5 November 2011 and 17 November 2011 in hibernacula in Michigan or Illinois, USA (M. lucifugus) and in Iowa or Illinois, USA (E. fuscus), as previously described (Moore et al. 2018). At the time of capture, none of these populations was known to have previous exposure to WNS. The bats were transported while torpid to Bucknell University in Lewisburg, Pennsylvania, USA where they were treated with either phosphate-buffered saline with 0.5% TWEEN-20 (control) or with a suspension of 350,000 Pd conidia (Pd exposed). Control and treated bats were separately housed in environmental chambers at 4°C and 95% relative humidity with each species in separate enclosures within the chamber. Following 3, 7, or 13 weeks of hibernation, bats were removed from the chamber and humanely euthanized by isoflurane overdose followed by decapitation. For the current study, the analysis was based on disease severity rather than hibernation time and time points were combined. The WAT was dissected from between the pelvis and the skin on both sides of the back, among other tissues, and snap-frozen in liquid nitrogen. Tissues were stored at -80°C to minimize enzymatic activity until they were processed for analysis.
Sample Type:Adipose tissue

Treatment:

Treatment ID:TR003573
Treatment Summary:The WAT samples used in this study were collected as part of a captive hibernation experiment performed at Bucknell University from November 2011 to March 2013. The bats were captured between 5 November 2011 and 17 November 2011 in hibernacula in Michigan or Illinois, USA (M. lucifugus) and in Iowa or Illinois, USA (E. fuscus), as previously described (Moore et al. 2018). At the time of capture, none of these populations was known to have previous exposure to WNS. The bats were transported while torpid to Bucknell University in Lewisburg, Pennsylvania, USA where they were treated with either phosphate-buffered saline with 0.5% TWEEN-20 (control) or with a suspension of 350,000 Pd conidia (Pd exposed). Control and treated bats were separately housed in environmental chambers at 4°C and 95% relative humidity with each species in separate enclosures within the chamber. Following 3, 7, or 13 weeks of hibernation, bats were removed from the chamber and humanely euthanized by isoflurane overdose followed by decapitation. For the current study, the analysis was based on disease severity rather than hibernation time and time points were combined. The WAT was dissected from between the pelvis and the skin on both sides of the back, among other tissues, and snap-frozen in liquid nitrogen. Tissues were stored at -80°C to minimize enzymatic activity until they were processed for analysis.

Sample Preparation:

Sampleprep ID:SP003571
Sampleprep Summary:Briefly, ~10 mg of tissue was homogenized as above, with tissue disruption after a 12-hr period at -80 °C, lipid isolation/purification using a liquid-liquid extraction, and lyophilization. The dried extract was reconstituted in 200 μL of extraction buffer (IPA) and filtered using a 0.2 μM microcentrifuge filter. The supernatant was transferred to MS vial for LC-MS analysis.

Combined analysis:

Analysis ID AN005647 AN005648
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Shimadzu Nexera X2 Shimadzu Nexera X2
Column Waters XBridge Amide (100 x 4.6mm,3.5um) Waters XBridge Amide (100 x 4.6mm,3.5um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap
Ion Mode POSITIVE NEGATIVE
Units peak area peak area

Chromatography:

Chromatography ID:CH004286
Chromatography Summary:Nexera X2 SIL-30 AC auto sampler (Shimazdu) connected with a high flow LC-30AD solvent delivery unit (Shimazdu) and Exion 30AD communication bus module (Shimazdu)
Instrument Name:Shimadzu Nexera X2
Column Name:Waters XBridge Amide (100 x 4.6mm,3.5um)
Column Temperature:35
Flow Gradient:The mobile phase was initially 100% solvent A, then a gradient of 3.0 min (solvent A 99.9%, B 0.01%), 3.0 min (solvent A 94%, B 6%), 4.0 min (solvent A 25%, B 75%), 6.0 min (solvent A 0%, B 100%), and then 6.0 min equilibration back to 100% solvent A
Flow Rate:0.7 mL/min
Solvent A:95% acetonitrile/5% water; 10 mM ammonium acetate
Solvent B:50% acetonitrile/50% water; 10 mM ammonium acetate
Chromatography Type:HILIC
  
Chromatography ID:CH004287
Chromatography Summary:Nexera X2 SIL-30 AC auto sampler (Shimazdu) connected with a high flow LC-30AD solvent delivery unit (Shimazdu) and Exion 30AD communication bus module (Shimazdu)
Instrument Name:Shimadzu Nexera X2
Column Name:Waters XBridge Amide (100 x 4.6mm,3.5um)
Column Temperature:35
Flow Gradient:The mobile phase was initially 100% solvent A, then a gradient of 3.0 min (solvent A 99.9%, B 0.01%), 3.0 min (solvent A 94%, B 6%), 4.0 min (solvent A 25%, B 75%), 6.0 min (solvent A 0%, B 100%), and then 6.0 min equilibration back to 100% solvent A
Flow Rate:0.7 mL/min
Solvent A:95% acetonitrile/5% water; 10 mM ammonium acetate
Solvent B:50% acetonitrile/50% water; 10 mM ammonium acetate
Chromatography Type:HILIC

MS:

MS ID:MS005371
Analysis ID:AN005647
Instrument Name:ABI Sciex 5500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:the qlm file was imported into MultiQuant v 2.0
Ion Mode:POSITIVE
  
MS ID:MS005372
Analysis ID:AN005648
Instrument Name:ABI Sciex 5500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:the qlm file was imported into MultiQuant v 2.0
Ion Mode:NEGATIVE
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