Summary of Study ST000451

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR000349. The data can be accessed directly via it's Project DOI: 10.21228/M8XS4Q This work is supported by NIH grant, U2C- DK119886.


Perform statistical analysis  |  Show all samples  |  Show named metabolites  |  Download named metabolite data  
Download mwTab file (text)   |  Download mwTab file(JSON)
Study IDST000451
Study TitleThe alpha-1A adrenergic receptor agonist A61603 reduces cardiac polyunsaturated fatty acid-Heart raw data
Study TypeGC-MS non-targeted metabolomic profiling
Study SummaryStudies of skeletal muscle disuse either in patients on bed rest or experimentally in animals(immobilization) have demonstrated that decreased protein synthesis is common, with transient parallel increases in protein degradation. Muscle disuse atrophy involves a process of transition from slow to fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for fat oxidation, and substrate accumulation in atrophied muscles have been reported as has accommodation of the liver with an increased gluconeogenic capacity. Recent studies have modeled skeletal muscle disuse by using cyclic stretch of differentiated myotubes (C2C12), which mimics the loading pattern of mature skeletal muscle, followed by cessation of stretch.We utilized this model to determine the metabolic changes using non-targeted metabolomics analysis of the media. We identified increases in amino acids resulting from protein degradation (largely sarcomere) that occurs with muscle atrophy that are involved in feeding the Kreb’s cycle through anaplerosis. Specifically, we identified increased alanine/proline metabolism (significantly elevated proline, alanine, glutamine, and asparagine) and increased -ketoglutaric acid, the proposed Kreb’s cycle intermediate being fed by the alanine/proline metabolic anaplerotic mechanism. Additionally, several unique pathways not clearly delineated in previous studies of muscle unloading were seen, including: 1) elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and 2-keovaline) represent intermediates in the Ehlrich amino acid degradation pathway, which feeds into a metabolic pathway supplying acetyl-CoA and 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an intermediate of purine metabolism, was seen at 12 hours unloading. Given the interest in targeting different aspects of the ubiquitin proteasome system to inhibit protein degradation, this C2C12 system may allow the identification of direct and indirect alterations in metabolism due to anaplerosis or through other yet to be identified mechanisms using a non-targeted metabolomics approach.
University of North Carolina at Chapel Hill
DepartmentMcAllister Heart Institute
LaboratoryMutliple Centers
Last NameIlaiwy;WIllis
First NameAmro;Monte
Address111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA,
Submit Date2016-08-17
Analysis Type DetailGC-MS
Release Date2016-09-23
Release Version1
Amro Ilaiwy Amro Ilaiwy
Monte WIllis Monte WIllis application/zip

Select appropriate tab below to view additional metadata details:


Project ID:PR000349
Project DOI:doi: 10.21228/M8XS4Q
Project Title:The alpha-1A adrenergic receptor agonist A61603 reduces cardiac polyunsaturated fatty acid
Project Type:GC-MS non targeted qualitative analysis
Project Summary:Introduction Alpha-1-adrenergic receptors (α1-ARs) are G-protein coupled receptors (GPCRs) with three highly homologous subtypes (α1A, α1B, and α1D). Of these three subtypes, only the α1A and α1B are expressed in the heart. Multiple pre-clinical models of heart injury demonstrate cardioprotective roles for the α1A. Non-selective α1-AR activation promotes glycolysis in the heart, but the functional α1-AR subtype and broader metabolic effects have not been studied. Objectives Given the high metabolic demands of the heart and previous evidence indicating benefit from α1A activation, we chose to investigate the effects of α1A activation on the cardiac metabolome in vivo. Methods Mice were treated for 1 week with a low, subpressor dose of A61603, a highly selective and potent α1A agonist. Cardiac tissue and serum were analyzed using a non-targeted metabolomics approach. Results We identified previously unrecognized metabolic responses to α1A activation,most notably broad reduction in the abundance of polyunsaturated fatty acids(PUFAs) and endocannabinoids (ECs). Conclusion Given the well characterized roles of PUFAs and ECs in inflammatory pathways, these findings suggest a possible role for cardiac α1A-ARs in the regulation of inflammation and may offer novel insight into the mechanisms underlying the cardioprotective benefit of selective pharmacologic α1A activation.
Institute:University of North Carolina at Chapel Hill
Department:McAllister heart Institute, Department of Internal medicine
Laboratory:Multiple Centers
Last Name:Ilaiwy; Willis
First Name:Amro; Monte
Address:111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA,
Funding Source:NIH, Fondation Leducq


Subject ID:SU000472
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammal


Subject type: Animal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
Showing results 1 to 16 of 16


Collection ID:CO000466
Collection Summary:Mice were sacrificed under deep isoflurane anesthesia, blood was collected by cardiac puncture, clotted, and 100–200 μL serum was snap frozen in liquid nitrogen. The heart was dissected, the atria and great vessels were excised,and the ventricles were allowed to beat in cold PBS to clear blood.
Sample Type:Muscle


Treatment ID:TR000486
Treatment Summary:N/A

Sample Preparation:

Sampleprep ID:SP000479
Sampleprep Summary:sample extract reconstituted in solvents containing a series of standards at fixed concentrations to ensure injection and chromatographic consistency. One aliquot was analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. The extract was gradient eluted from a C18 column (Waters UPLC BEH C18-2.1 × 100 mm, 1.7 μm) using water and methanol, containing 0.05 % perfluoropentanoic acid (PFPA) and 0.1 % formic acid (FA). Another aliquot was also analyzed using acidic positive ion conditions,however it was chromatographically optimized for more hydrophobic compounds. In this method, the extract was gradient eluted from the same C18 column using methanol, acetonitrile, water, 0.05 % PFPA and 0.01 % FA and was operated at an overall higher organic content. Another aliquot was analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts were gradient eluted from the column using methanol and water

Combined analysis:

Analysis ID AN000707
Analysis type MS
Chromatography type GC
Chromatography system Waters Acquity
Column Waters Acquity BEH C18 (100 x 2mm,1.7um)
MS Type EI
MS instrument type Single quadrupole
MS instrument name Agilent 5975
Units Peak values (scaled)


Chromatography ID:CH000511
Instrument Name:Waters Acquity
Column Name:Waters Acquity BEH C18 (100 x 2mm,1.7um)
Chromatography Type:GC


MS ID:MS000629
Analysis ID:AN000707
Instrument Name:Agilent 5975
Instrument Type:Single quadrupole
MS Type:EI