Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics
Stable isotope-resolved metabolomics (SIRM) is a powerful tool for understanding disease. Advances in SIRM techniques have improved isotopic delivery and expanded the workflow from exclusively in vitro applications to in vivo methodologies to study systemic metabolism. Here, we report a simple, mini...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-12-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/10/12/501 |
| _version_ | 1827700532655947776 |
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| author | Holden C. Williams Margaret A. Piron Grant K. Nation Adeline E. Walsh Lyndsay E. A. Young Ramon C. Sun Lance A. Johnson |
| author_facet | Holden C. Williams Margaret A. Piron Grant K. Nation Adeline E. Walsh Lyndsay E. A. Young Ramon C. Sun Lance A. Johnson |
| author_sort | Holden C. Williams |
| collection | DOAJ |
| description | Stable isotope-resolved metabolomics (SIRM) is a powerful tool for understanding disease. Advances in SIRM techniques have improved isotopic delivery and expanded the workflow from exclusively in vitro applications to in vivo methodologies to study systemic metabolism. Here, we report a simple, minimally-invasive and cost-effective method of tracer delivery to study SIRM in vivo in laboratory mice. Following a brief fasting period, we orally administered a solution of [U-<sup>13</sup>C] glucose through a blunt gavage needle without anesthesia, at a physiological dose commonly used for glucose tolerance tests (2 g/kg bodyweight). We defined isotopic enrichment in plasma and tissue at 15, 30, 120, and 240 min post-gavage. <sup>13</sup>C-labeled glucose peaked in plasma around 15 min post-gavage, followed by period of metabolic decay and clearance until 4 h. We demonstrate robust enrichment of a variety of central carbon metabolites in the plasma, brain and liver of C57/BL6 mice, including amino acids, neurotransmitters, and glycolytic and tricarboxylic acid (TCA) cycle intermediates. We then applied this method to study in vivo metabolism in two distinct mouse models of diseases known to involve dysregulation of glucose metabolism: Alzheimer’s disease and type II diabetes. By delivering [U-<sup>13</sup>C] glucose via oral gavage to the 5XFAD Alzheimer’s disease model and the Lep<sup>ob/ob</sup> type II diabetes model, we were able to resolve significant differences in multiple central carbon pathways in both model systems, thus providing evidence of the utility of this method to study diseases with metabolic components. Together, these data clearly demonstrate the efficacy and efficiency of an oral gavage delivery method, and present a clear time course for <sup>13</sup>C enrichment in plasma, liver and brain of mice following oral gavage of [U-<sup>13</sup>C] glucose—data we hope will aid other researchers in their own <sup>13</sup>C-glucose metabolomics study design. |
| first_indexed | 2024-03-10T14:14:26Z |
| format | Article |
| id | doaj.art-3f3d40d074544e3fa09a3c8ed3e8da6a |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-10T14:14:26Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-3f3d40d074544e3fa09a3c8ed3e8da6a2023-11-20T23:53:04ZengMDPI AGMetabolites2218-19892020-12-01101250110.3390/metabo10120501Oral Gavage Delivery of Stable Isotope Tracer for In Vivo MetabolomicsHolden C. Williams0Margaret A. Piron1Grant K. Nation2Adeline E. Walsh3Lyndsay E. A. Young4Ramon C. Sun5Lance A. Johnson6Department of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY 40536, USASanders-Brown Center on Aging, University of Kentucky College of Medicine, Lexington, KY 40536, USADepartment of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USAStable isotope-resolved metabolomics (SIRM) is a powerful tool for understanding disease. Advances in SIRM techniques have improved isotopic delivery and expanded the workflow from exclusively in vitro applications to in vivo methodologies to study systemic metabolism. Here, we report a simple, minimally-invasive and cost-effective method of tracer delivery to study SIRM in vivo in laboratory mice. Following a brief fasting period, we orally administered a solution of [U-<sup>13</sup>C] glucose through a blunt gavage needle without anesthesia, at a physiological dose commonly used for glucose tolerance tests (2 g/kg bodyweight). We defined isotopic enrichment in plasma and tissue at 15, 30, 120, and 240 min post-gavage. <sup>13</sup>C-labeled glucose peaked in plasma around 15 min post-gavage, followed by period of metabolic decay and clearance until 4 h. We demonstrate robust enrichment of a variety of central carbon metabolites in the plasma, brain and liver of C57/BL6 mice, including amino acids, neurotransmitters, and glycolytic and tricarboxylic acid (TCA) cycle intermediates. We then applied this method to study in vivo metabolism in two distinct mouse models of diseases known to involve dysregulation of glucose metabolism: Alzheimer’s disease and type II diabetes. By delivering [U-<sup>13</sup>C] glucose via oral gavage to the 5XFAD Alzheimer’s disease model and the Lep<sup>ob/ob</sup> type II diabetes model, we were able to resolve significant differences in multiple central carbon pathways in both model systems, thus providing evidence of the utility of this method to study diseases with metabolic components. Together, these data clearly demonstrate the efficacy and efficiency of an oral gavage delivery method, and present a clear time course for <sup>13</sup>C enrichment in plasma, liver and brain of mice following oral gavage of [U-<sup>13</sup>C] glucose—data we hope will aid other researchers in their own <sup>13</sup>C-glucose metabolomics study design.https://www.mdpi.com/2218-1989/10/12/501metabolomicsstable isotope<sup>13</sup>C-glucosegavageSIRMAlzheimer’s disease |
| spellingShingle | Holden C. Williams Margaret A. Piron Grant K. Nation Adeline E. Walsh Lyndsay E. A. Young Ramon C. Sun Lance A. Johnson Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics Metabolites metabolomics stable isotope <sup>13</sup>C-glucose gavage SIRM Alzheimer’s disease |
| title | Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics |
| title_full | Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics |
| title_fullStr | Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics |
| title_full_unstemmed | Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics |
| title_short | Oral Gavage Delivery of Stable Isotope Tracer for In Vivo Metabolomics |
| title_sort | oral gavage delivery of stable isotope tracer for in vivo metabolomics |
| topic | metabolomics stable isotope <sup>13</sup>C-glucose gavage SIRM Alzheimer’s disease |
| url | https://www.mdpi.com/2218-1989/10/12/501 |
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