Summary of Study ST002356

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 PR001513. The data can be accessed directly via it's Project DOI: 10.21228/M8471X This work is supported by NIH grant, U2C- DK119886.

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Study IDST002356
Study TitleIsolated murine skeletal muscles utilize pyruvate over glucose for oxidation-Part 1
Study TypeStudy of the different substrate by isolated skeletal muscle at room temperature via C-13 isotopomer analysis
Study SummaryPreclinical studies of muscle contractile function often employ ex vivo preparations of the soleus and/or extensor digitorum longus (EDL) muscles which are relatively easy to prepare and represent slow and fast fiber properties, respectively. Therefore, the current study sought to examine the utility of this preparation for understanding the metabolic fuel utilization in isolated resting mouse muscles at room temperature. 13C-labeling in both muscle types was performed using three fuels: glucose, pyruvate, and acetate, followed by NMR-based metabolomics analyses. Incubating 13C-labeled substrates in the isolated skeletal muscles makes it possible to examine TCA cycle flux and substrate selection by these muscles.
Institute
University of Florida
DepartmentApplied Physiology and Kinesiology
LaboratoryRm 42 and Rm 43
Last NameKhattri
First NameRam
Address1864 Stadium RD, Gainesville, FL, 32611, USA
Emailrbk11@ufl.edu
Phone3307856045
Submit Date2022-10-31
Num Groups4
Total Subjects18
Study CommentsSoutheastern Center for Integrated Metabolomics (SECIM) (ERB), NIH AR U54 AR052646 (Physiological Assessment Core, ERB), and Wellstone Muscular Dystrophy Cooperative Research Center Grant (NIAMS: U54AR052646/P50 AR052646). The AMRIS Facility is supported by the National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida.
Raw Data AvailableYes
Raw Data File Type(s)fid
Analysis Type DetailNMR
Release Date2023-05-01
Release Version1
Ram Khattri Ram Khattri
https://dx.doi.org/10.21228/M8471X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Collection:

Collection ID:CO002438
Collection Summary:Mice were anesthetized (using a combination of xylazine (80mg/kg) and ketamine (10mg/kg)) to allow removal of soleus and extensor digitorum longus (EDL) muscles. Upon removal, muscles were incubated at 22 oC in Ringer/MEM solution gas equilibrated with 95/5% O2/CO2 with appropriate 13C labeled substrates in a perfusion chamber routinely used for isolated muscle mechanics for 30 minutes. These included the following: 5.5 mM [U-13C6] glucose; 5.5 mM [U-13C3] pyruvate, or 16.5 mM [13C2] labeled Na-acetate. Following incubation, muscles were quickly removed, blotted, and then rapidly frozen in liquid nitrogen for subsequent NMR analysis.
Collection Protocol Comments:Mice were anesthetized (using a combination of xylazine (80mg/kg) and ketamine (10mg/kg)) to allow removal of soleus and extensor digitorum longus (EDL) muscles. Upon removal, muscles were incubated at 22 oC in Ringer/MEM solution gas equilibrated with 95/5% O2/CO2 with appropriate 13C labeled substrates in a perfusion chamber routinely used for isolated muscle mechanics for 30 minutes. These included the following: 5.5 mM [U-13C6] glucose; 5.5 mM [U-13C3] pyruvate, or 16.5 mM [13C2] labeled Na-acetate. Following incubation, muscles were quickly removed, blotted, and then rapidly frozen in liquid nitrogen for subsequent NMR analysis. N=4 muscles were pooled into a single biological replicate of 30-50 mg tissue to afford detectable levels of substrates in the NMR analysis.
Sample Type:Muscle
Collection Method:Mice were anesthetized (using a combination of xylazine (80mg/kg) and ketamine (10mg/kg)) to allow removal of soleus and extensor digitorum longus (EDL) muscles.
Collection Location:University of Florida, Applied Physiology and Kinesiology, MBI, 1149 Newell Dr, Gainesville, FL 32610
Storage Conditions:-80℃
Collection Vials:cryovials
Storage Vials:cryovials
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