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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:Glucose tolerance test with insulin secretion measures systemic clearance of glucose and systemic appearance of insulin following an intraperitoneal bolus injection of 20% dextrose. This experiment measures insulin sensitivity and insulin secretion (pancreatic -cell function) in awake mice assuming that there are no alterations in systemic insulin clearance. Insulin sensitivity and pancreatic -cell function are altered in obese mice.
Proper citation: Jason Kim 2019. U Mass - Glucose Tolerance Test with insulin secretion. protocols.io dx.doi.org/10.17504/protocols.io.xxbfpin Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary:
This experiment measures serum and plasma concentrations of non-esterified fatty acids using a 96-well kit. The experiment involves a coupled reaction to measure non-esterified fatty acids (NEFA) which ultimately forms a purple product that absorbs light at 550nm. This allows the concentration of NEFA to be determined from the optical density measured at 540~550nm. Serum fatty acids levels reflect systemic lipid metabolism, lipid digestion/absorption, and lipid clearance. Serum fatty acids levels are altered in obesity, insulin resistance, and type 2 diabetes.
Proper citation: Jason Kim 2019. U Mass - Non-esterified fatty acids. protocols.io dx.doi.org/10.17504/protocols.io.x3hfqj6 Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum levels of total protein reflect systemic protein metabolism.
Proper citation: Jason Kim 2019. U Mass - Total protein. protocols.io dx.doi.org/10.17504/protocols.io.x5gfq3w Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary:
The EchoMRI 3-in-1 uses ¹H- magnetic resonance spectroscopy to noninvasively measure fat mass, lean mass, and water mass in individual organs. Fat, lean, and water mass composition in specific organs is altered in obesity.
Proper citation: Jason Kim 2019. U Mass - Body composition (organs). protocols.io dx.doi.org/10.17504/protocols.io.xsnfnde Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers, Metabolomics Protocols & Workflows
Summary: This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Triglyceride levels may be measured in serum, plasma, and tissue samples. Serum and tissue triglyceride levels are affected by alterations in systemic lipid metabolism, lipid digestion/absorption, and lipid clearance. Serum and tissue triglyceride levels are altered in obesity, insulin resistance, type 2 diabetes, alcoholic steatohepatitis, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis.
Proper citation: Jason Kim 2019. U Mass - Triglyceride. protocols.io dx.doi.org/10.17504/protocols.io.x5pfq5n Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:Phloridzin is a potent inhibitor of renal glucose reabsorption and may be used to lower serum glucose levels. Phloridzin may be administered chronically to induce glycosuria and lower hyperglycemia in diabetic mice. Phloridzin may be administered acutely to reduce serum glucose levels for experiments in diabetic mice.
Proper citation: Jason Kim 2019. U Mass - Chronic/acute phloridzin treatment. protocols.io dx.doi.org/10.17504/protocols.io.xujfnun Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
This experiment provides the quantification of multiple cytokines and chemokines using multiplexed-Luminex technology based on beads containing specific antibodies. Serum cytokine levels reflect chronic or acute inflammation, and circulating cytokines and chemokines are altered in obesity.
Cytokines Panel I include IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, KC (IL-8 homologue), IL-9, IL-10 (interleukin-10), IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, TNFα (tumor necrosis factor alpha), IFNγ (interferon gamma), IP-10 (interferon gamma-induced protein 10; CXCL-10), Eotaxin (CCL-11), G-CSF (granulocyte colony stimulating factor), GM-CSF (granulocyte macrophage colony stimulating factor), LIF (leukemia inhibitory factor), LIX (LPS- induced CXC chemokine), MCP-1 (monocyte chemotactic protein-1; CCL-2), M-CSF (macrophage colony stimulating factor), MIG (monokine induced by gamma interferon; CXCL- 9), MIP-1α (macrophage inflammatory protein alpha; CCL-3), MIP-1 (macrophage inflammatory protein beta; CCL-4), and MIP-2/RANTES (macrophage inflammatory protein 2- alpha; CXCL-2).
A service can be requested for all or any combination of listed cytokines/chemokines for customized multiplexed Luminex assay.
Proper citation: Jason Kim 2019. U Mass - Cytokines Panel I - multiplex. protocols.io dx.doi.org/10.17504/protocols.io.xvnfn5e Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
Whole body glucose turnover and hepatic glucose production rates are measured at basal state using an intravenous infusion of labeled glucose in awake mice. Whole body glucose turnover and hepatic glucose production regulate basal glucose levels and are altered in obesity.
Proper citation: Jason Kim 2019. U Mass - Basal glucose metabolism. protocols.io dx.doi.org/10.17504/protocols.io.xr8fm9w Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Hyperinsulinemic-euglycemic clamp is the gold-standard method to assess insulin sensitivity. The hyperinsulinemic-euglycemic clamp is widely used in clinics and laboratories to measure insulin action on glucose utilization in humans and animals for clinical and basic science research. Incorporation of radioactive-labeled glucose during hyperinsulinemic-euglycemic clamps makes it possible to measure glucose metabolism in individual organs in awake mice. Impaired insulin sensitivity (insulin resistance) is a major characteristic of obesity and an early requisite event in the development of type 2 diabetes.
Proper citation: Jason Kim 2019. U Mass - Protein metabolism. protocols.io dx.doi.org/10.17504/protocols.io.x4ffqtn Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: A high-fat diet of varying composition and percent fat is administered to induce obesity in mice. High-fat diet induced obesity is causally associated with insulin resistance and type 2 diabetes.
Proper citation: Jason Kim 2019. U Mass - Chronic high-fat feeding. protocols.io dx.doi.org/10.17504/protocols.io.xugfntw Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum levels of bilirubin reflect bile acid metabolism.
Proper citation: Jason Kim 2019. U Mass - Bilirubin. protocols.io dx.doi.org/10.17504/protocols.io.xscfnaw Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary:
The EchoMRI 3-in-1 uses ¹H- magnetic resonance spectroscopy to noninvasively measure whole body fat mass, lean mass, and water mass in awake mice. Whole body fat mass is a measure of adiposity that is altered in obesity. Whole body lean mass may be altered by exercise and in mouse models of altered metabolism.
Proper citation: Jason Kim 2019. U Mass - Body composition (whole body). protocols.io dx.doi.org/10.17504/protocols.io.xsqfndw Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:The TSE PhenoMaster/LabMaster Metabolic Cage system is used to measure indirect calorimetry, food/water intake, energy expenditure, and physical activity in individually housed awake mice. The experiment noninvasively measures VO² consumption and VCO² production rates in individual mice using metabolic chambers and calculates the respiratory exchange ratio (respiratory quotient) to reflect energy expenditure. Metabolic cage measurement is conducted continuously for 72 hours (3 days) to account for acclimation of mice housed in home cages during the study. With our high-speed Siemens O²/CO² sensing unit, indirect calorimetry measurements may be performed at a 20-min interval for a full 12-cage study.
Proper citation: Jason Kim 2019. U Mass - Energy balance – food intake, energy expenditure, physical activity. protocols.io dx.doi.org/10.17504/protocols.io.xwwfpfe Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Chronic indwelling catheter is placed in the carotid artery for blood sampling during experiments. Surival surgery is performed in anesthetized mice, and mice recover from surgery after 4~5 days.
Proper citation: Jason Kim 2019. U Mass - Surgery – carotid artery cannulation. protocols.io dx.doi.org/10.17504/protocols.io.x4pfqvn Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary:
This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Lipase is a gastrointestinal enzyme that processes fat/lipid digestion.
Proper citation: Jason Kim 2019. U Mass - Lipase. protocols.io dx.doi.org/10.17504/protocols.io.x3bfqin Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
This experiment provides the quantification of multiple cytokines and chemokines using multiplexed-Luminex technology based on beads containing specific antibodies. Serum cytokine levels reflect chronic or acute inflammation, and circulating cytokines and chemokines are altered in obesity.
Cytokines Panel II include IL-16 (interleukin-16), IL-17E/IL-25, IL-21, IL-22, IL-28B, EPO (erythropoietin), Exodus-2 (CCL-21), Fractalkine (CX3CL1), MCP-5 (monocyte chemotactic protein-5; CCL-12), MIP-3α (macrophage inflammatory protein 3-alpha; CCL-20), MIP-3β(macrophage inflammatory protein 3-beta; CCL-19), and TARC (thymus and activation- regulated; CCL-17).
A service can be requested for all or any combination of listed cytokines/chemokines for customized multiplexed Luminex assay.
Proper citation: Jason Kim 2019. U Mass - Cytokines Panel II - multiplex. protocols.io dx.doi.org/10.17504/protocols.io.xvvfn66 Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Triglyceride emulsion and heparin will be intravenously infused for 5 hours to acutely raise circulating fatty acids levels in awake mice. Acute lipid infusion is shown to cause insulin resistance in peripheral organs.
Proper citation: Jason Kim 2019. U Mass - Acute lipid infusion. protocols.io dx.doi.org/10.17504/protocols.io.xm5fk86 Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary:
This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer.
Proper citation: Jason Kim 2019. U Mass - Lactate dehydrogenase. protocols.io dx.doi.org/10.17504/protocols.io.x2yfqfw Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum levels of C-reactive peptide reflect systemic inflammation and may be altered in obesity.
Proper citation: Jason Kim 2019. U Mass - C-reactive Peptide. protocols.io dx.doi.org/10.17504/protocols.io.xt5fnq6 Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Streptozotocin can selectively destroy the pancreatic β-cells with rapid and irreversible necrosis and can be used to generate a chronic model of hyperglycemia and type 1 diabetes.
Proper citation: Jason Kim 2019. U Mass - STZ-induced type 1 diabetes model. protocols.io dx.doi.org/10.17504/protocols.io.xh8fj9w Copy
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