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Authors: David A. Antonetti
Group: Mouse Metabolic Phenotyping Centers
Summary: A virtual optometry system is used to quantify the spatial vision of laboratory animal.
Proper citation: David A. Antonetti 2019. U Michigan - Optokinetic Measurements of Visual Acuity and Contrast Sensitivity. protocols.io dx.doi.org/10.17504/protocols.io.x99fr96 Copy
Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:Chronic indwelling catheter is placed in the jugular vein for intravenous infusion and injection of drug, hormones, and metabolites. Surival surgery is performed in anesthetized mice, and mice recover from surgery after 4~5 days.
Proper citation: Jason Kim 2019. U Mass - Surgery – jugular vein cannulation. protocols.io dx.doi.org/10.17504/protocols.io.x5cfq2w Copy
Authors: Kristin Evans
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
Vertical sleeve Gastrectomy (VSG) is a common surgical treatment of obesity, type-2 diabetes, and other comorbidities of obesity. The VSG mouse surgical model provides researchers with a mechanism to evaluate mechanistic studies to explore the methods by which VSG bariatric surgery produces beneficial effects on obesity. The surgical procedure is performed under sterile conditions in accordance with AAALAC standards. Surgical animals will be able to ship 2 weeks after the post-operative monitoring period is completed. Cohorts of Vertical Sleeve gastrectomy (VSG), a pair fed SHAM, and an ad lib SHAM will be provided with each cohort ordered.
Proper citation: Kristin Evans 2019. UC Davis - Vertical Sleeve Gastrectomy. protocols.io dx.doi.org/10.17504/protocols.io.za3f2gn Copy
Authors: Patrick Tso, Dana Lee
Group: Mouse Metabolic Phenotyping Centers
Summary: Total body composition in live, un-anaesthetized small animals and carcasses will reveal absolute amounts of body fat, lean tissue and body water via a quantitative magnetic resonance (QMR) instrument, EchoMRI, (Echo Medical Systems, LLC, Houston, TX). This instrument uses the differences in the nuclear magnetic resonance properties of hydrogen atoms in organic and non-organic environments to fractionate signals originating from fat, lean tissue and free water.
Proper citation: Patrick Tso, Dana Lee 2019. U Cinn - Body Composition & Carcass Analysis. protocols.io dx.doi.org/10.17504/protocols.io.2qigdue Copy
Authors: Louise Lantier
Group: Mouse Metabolic Phenotyping Centers
Summary: The glucose tolerance test measures the clearance of an intraperitoneally injected glucose load from the body. It is used to detect disturbances in glucose metabolism that can be linked to diabetes or metabolic syndrome. Animals are fasted for 5 hours, fasted blood glucose levels are determined before a solution of glucose is administered by intra-peritoneal (IP) injection. Subsequently, the blood glucose level is measured at different time points during the following 90 minutes.
Proper citation: Louise Lantier 2019. Vandy – IP glucose tolerance test. protocols.io dx.doi.org/10.17504/protocols.io.yzzfx76 Copy
Authors: Richard Mcindoe, Mark Kelly, Timothy P. Fitzgibbons
Group: Mouse Metabolic Phenotyping Centers
Summary: This is a mouse model of acute myocardial infarction. This protocol is suitable for studying mouse models thought to be relevant to coronary ischemia or the development of congestive heart failure.
Proper citation: Richard Mcindoe, Mark Kelly, Timothy P. Fitzgibbons 2019. UMass - Coronary Artery Ligation. protocols.io dx.doi.org/10.17504/protocols.io.wc4fayw Copy
Authors: Fawaz G. Haj
Group: Mouse Metabolic Phenotyping Centers
Summary: This test is designated to determine if rodents exhibit signs of endoplasmic reticulum stress, through evaluation of the activation state of the 3 sub-arms: PERK/EiF2α, Ire1α/sXBP1, and ATF6α pathways such. We will examine induction of ER stress in adipose and liver tissues.
Proper citation: Fawaz G. Haj 2019. UC Davis - Endoplasmic reticulum stress. protocols.io dx.doi.org/10.17504/protocols.io.yihfub6 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 - Creatine Kinase. protocols.io dx.doi.org/10.17504/protocols.io.xumfnu6 Copy
Authors: Peter Takeuchi, Trina Knotts
Group: Mouse Metabolic Phenotyping Centers
Summary: Energy expenditure is measured using an indirect respiration calorimetry system (CLAMS, Columbus Instruments, Columbus, OH). Air is drawn through calorimetry chambers and flow rate is controlled and measured with a mass flow controller. Oxygen consumption and carbon dioxide production are measured and used to calculate energy expenditure (or heat production, kilocalories (kcal)) and respiratory exchange ratio (RER: VCO2/VO2). The calorimetry chambers are also equipped with sensors, which allow the measurement of physical activity, food intake and water intake. All calorimetry chambers are housed within a temperature and light controlled incubator. The system contains 8 calorimetry chambers. Animals are acclimated to the facility for at least 1 week. Animals are then acclimated to the CLAMS cages and powdered diet for 48 hours and to the light and temperature-controlled chamber for 24 hours prior to testing. Analyzed data constitutes data collected from 48 hours of continuous measurement (2 light/2 dark cycles). To investigate the influence of exercise on energy expenditure and substrate oxidation, the chambers can also be fitted with running wheels or the animals can be placed in separate chambers containing treadmills.
Proper citation: Peter Takeuchi, Trina Knotts 2019. UC Davis - Comprehensive Laboratory Animal Monitoring System (CLAMS). protocols.io dx.doi.org/10.17504/protocols.io.yggfttw Copy
Authors: Jeff Hodgin
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:Collection of urine from experimental animals is a basic requirement to detect albumin/creatinine ratio for glomerular nephropathy. This is a non-invasive, simple, quick, and inexpensive method for collecting spot urine from mouse when a small volume of urine (10-500ul) sample is required.
Proper citation: Jeff Hodgin 2019. U Michigan - Spot urine collection. protocols.io dx.doi.org/10.17504/protocols.io.56fg9bn Copy
Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
The enzyme methodology employed in this reagent is based on the reaction first described by Talke and Schubert. To shorten and simplify the assay, the calculations are based on the discovery of Tiffany, et al. that urea concentration is proportional to absorbance change over a fixed time interval. The reaction is monitored by measuring the rate of decrease in absorbance at 340 nm as NADH is converted to NAD.
Proper citation: Peter Havel 2019. UC Davis - Urea Protocol. protocols.io dx.doi.org/10.17504/protocols.io.yw6fxhe Copy
Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: The method employed here is based on an enzymatic colorimetric determination of creatinine which largely eliminates interferences known to the Jaffe method. In the final reaction sequence, the formation of the quinoneimine dye product results in an increase in absorbance at 550 nm (530-570 nm) which is directly proportional to the creatinine concentration in the sample. Potential interference from endogenous creatine and sarcosine are eliminated by the reaction of creatine amidinohydrolase, sarcosine oxidase and catalase before creatinine is determined. Ascorbate oxidase is included in the reagent to eliminate the influence of ascorbate in the sample.
Proper citation: Peter Havel 2019. UC Davis - Creatinine Protocol. protocols.io dx.doi.org/10.17504/protocols.io.ygjftun Copy
Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
Glutathione peroxidase (GPx) catalyzes the reduction of hydroperoxides, including hydrogen peroxides, by reduced glutathione and functions to protect the cell from oxidative damage. With the exception of phospholipid-hydroperoxide GPx, a monomer, all of the GPx enzymes are tetramers of four identical subunits. Each subunit contains a selenocysteine in the active site which participates directly in the two-electron reduction of the peroxide substrate. The enzyme uses glutathione as the ultimate electron donor to regenerate the reduced form of the selenocysteine. The Cayman Chemical Glutathione Peroxidase Assay Kit measures GPx activity indirectly by a coupled reaction with glutathione reductase (GR). Oxidized glutathione (GSSG), produced upon reduction of an organic hydroperoxide by GPx, is recycled to its reduced state by GR and NADPH. The oxidation of NADPH to NADP+ is accompanied by a decrease in absorbance at 340 nm. The rate of decrease in the A340 is directly proportional to the GPx activity in the sample. The Cayman GPx Assay Kit can be used to measure all of the glutathione- dependent peroxidases in plasma, erythrocyte lysates, tissue homogenates, and cell lysates.
Proper citation: Peter Havel 2019. UC Davis - Glutathione Peroxidase. protocols.io dx.doi.org/10.17504/protocols.io.ykdfus6 Copy
Authors: Gary Cline, John Stack
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure to measure the amount of Alkaline Phosphatase activity. Alkaline Phosphatase (ALP) activity is measured from the hydrolysis of 4-nitrophenylphospate to 4-nitrophenyoxide ion (monitored at 405 nm) and phosphate.
Proper citation: Gary Cline, John Stack 2019. Yale - Alkaline Phosphatase. protocols.io dx.doi.org/10.17504/protocols.io.yz7fx9n Copy
Authors: K.C. Kent Lloyd
Group: Mouse Metabolic Phenotyping Centers
Summary: The purpose of this policy is to outline the minimum standards of care for rodents based on the Animal Welfare Act, the Public Health Service Policy, and the ILAR Guide for the Care and Use of Laboratory Animals. Specific procedures for this program can also be located in MBPVIV-20-105 Daily Obervations/Action and MBPVIV-31-101 Changing Mouse Cages. These procedures are to be followed by trained staff responsible for providing husbandry care for the mouse colony at MBP.
Proper citation: K.C. Kent Lloyd 2019. UC Davis - Husbandry Care for Mice. protocols.io dx.doi.org/10.17504/protocols.io.8nqhvdw Copy
Authors: John Stack, Gary Cline
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure to determine the total protein concentration in blood, serum, and plasma. Total Protein is determined by the biuret reaction.
Proper citation: John Stack, Gary Cline 2019. Yale - Total Protein. protocols.io dx.doi.org/10.17504/protocols.io.y4afyse Copy
Authors: John Rutledge
Group: Mouse Metabolic Phenotyping Centers
Summary: One index of arterial function that is compromised to a varying degree in individuals with cardiovascular disease is vascular stiffness. This protocol can be utilized to analyze vascular function as an index for cardiovascular disease. This protocol directly measures the stress and strain on an arterial vessel as an index of vascular function.Modified from: Symons et. al. Arterioscler Thromb Vasc Biol. 2002 May 1;22(5):772-80.
Proper citation: John Rutledge 2019. UC Davis - Maximum passive stiffness (MPS). protocols.io dx.doi.org/10.17504/protocols.io.yrtfv6n Copy
Authors: Mark Kelly, Timothy P. Fitzgibbons
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:
This is a Mouse model of Transverse aortic constriction (TAC). A commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart failure. Initially leading to compensated hypertrophy of the heart, which often is associated with a temporary enhancement of cardiac contractility. Resulting in cardiac dilatation and heart failure.
Proper citation: Mark Kelly, Timothy P. Fitzgibbons 2019. U Mass - Transverse Aortic Constriction. protocols.io dx.doi.org/10.17504/protocols.io.56ig9ce Copy
Authors: John Stack, Gary Cline
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure for determining the glycogen content within liver and muscle tissue.
Proper citation: John Stack, Gary Cline 2019. Yale - Tissue Glycogen. protocols.io dx.doi.org/10.17504/protocols.io.y39fyr6 Copy
Authors: Anil Singapuri
Group: Mouse Metabolic Phenotyping Centers
Summary: Cardiac hypertrophy is one of the most common causes of heart failure. The development of cardiac hypertrophy and failure can be monitored using noninvasive imaging with echocardiogram in conscious animals.Modified from: Timofeyev V. et al. J Mol Cell Cardiol. 2006 Jul;41(1):170-81.
Proper citation: Anil Singapuri 2019. UC Davis - Echocardiography. protocols.io dx.doi.org/10.17504/protocols.io.yiafuae Copy
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