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Authors: K.C. Kent Lloyd
Group: Mouse Metabolic Phenotyping Centers
Summary: The purpose of this document is to describe the procedure for accepting commercial vendor and courier-delivered (“Imports”) shipments of mice to the 2nd Street facility of the UC Davis Mouse Biology Program. This procedure is to be followed by administrative personnel responsible for accepting commercial or courier-delivered shipments of mice and by Vivarium personnel responsible for unpacking and caging mice. Transportation of mice is stressful and potentially dangerous to their health, especially in warm summer months. Mice need to be unpacked, have a health inspection, and housed in appropriate caging in appropriate numbers as soon as possible after arrival. This insures they are acclimated and in the best physical health prior to their use by the Mouse Biology Program.

Proper citation: K.C. Kent Lloyd 2019. UC Davis - Receipt of Vendor and Courier Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.xh4fj8w Copy   

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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 https://dx.doi.org/10.17504/protocols.io.yz7fx9n Copy   

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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 https://dx.doi.org/10.17504/protocols.io.8nqhvdw Copy   

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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 https://dx.doi.org/10.17504/protocols.io.yrtfv6n Copy   

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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 https://dx.doi.org/10.17504/protocols.io.y4afyse Copy   

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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 https://dx.doi.org/10.17504/protocols.io.xujfnun Copy   

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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 https://dx.doi.org/10.17504/protocols.io.yiafuae Copy   

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Authors: Jennifer Rutkowsky
Group: Mouse Metabolic Phenotyping Centers
Summary: This technique has been used routinely for the non-invasive measurement of blood pressure in rats, and more recently in mice. The technique provides a good estimate of actual systolic pressure. Hypertension major risk factor for stroke myocardial infarction, heart failure, aneurysm of the arteries, peripheral artery disease and is a cause of chronic kidney disease and erectile dysfunction (ED)Modified from: UC Davis MBP SOP-CODA Blood Pressure, Todd Tolentino 11/02/2012

Proper citation: Jennifer Rutkowsky 2019. UC Davis - Blood Pressure by Tail Cuff. protocols.io https://dx.doi.org/10.17504/protocols.io.yetften Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: A subcutaneous or intraperitoneal implantation of Alzet osmotic pump is used to chronically administer selected drug in mice. Chronic drug delivery may be used to examine intermediate to long-term effects of selected drug on obesity, insulin resistance, and metabolism.

Proper citation: Jason Kim 2019. U Mass - Chronic drug delivery. protocols.io https://dx.doi.org/10.17504/protocols.io.xuefnte Copy   

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Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: LDL and VLDL are separated from HDL using a precipitation reagent. Then the HDL fraction is measured for either TC or TG using the same reagents for total cholesterol or triglyceride.

Proper citation: Peter Havel 2019. UC Davis - LDL Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.yrcfv2w Copy   

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Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: The RIA (RadioImmuno Assay) is an assay method used for the quantification of various proteins. A standardized concentration of antibody specific to the analyte of interest is pipetted into test tubes. A standardized concentration of the analyte of interest that is labeled with a radioisotope (usually 125I) is added to the tubes. Standards and samples are pipetted into the tubes and the tubes are incubated. During the incubation period the standardized concentration of labeled analyte and the unknown concentration of analyte in the samples will compete for binding sites on the antibody. After the incubation period a precipitating reagent is added to the tubes and the bound antibodies are precipitated out using the double antibody-polyethyleneglycol precipitation technique. The tubes are centrifuged and the supernatant is aspirated and the precipitate is counted (using a gamma counter if 125I is used). If there is a low concentration of the analyte of interest in the samples, the labeled analyte will have a higher probability for binding to the antibody and thus there will be a higher count in the precipitate. If there is a high concentration of the analyte of interest in the samples there will be a lower count of labeled analyte in the precipitate. The counts and known concentrations of the standards are used to generate a standard curve, and the counts of the samples are used to interpolate quantitative concentrations for the analyte of interest from the standard curve.

Proper citation: Peter Havel 2019. UC Davis - Corticosterone RIA. protocols.io https://dx.doi.org/10.17504/protocols.io.ygiftue Copy   

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Authors: Fawaz G. Haj
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: An in-vivo glucose-stimulates insulin secretion test is designated to determine alterations in insulin secretion by the pancreas upon a bolus IP Glucose injection.

Proper citation: Fawaz G. Haj 2019. UC Davis - IN-VIVO Glucose-stimulates Insulin Secretion Test. protocols.io https://dx.doi.org/10.17504/protocols.io.yptfvnn Copy   

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Authors: Teri Stevenson, Vance L. Albaugh
Group: Mouse Metabolic Phenotyping Centers
Summary: This is the protocol for the modified gastric bypass in the mouse. In this procedure the stomach is bypassed and the food flows through the bypass arm directly into the jejunum.

Proper citation: Teri Stevenson, Vance L. Albaugh 2019. Vandy - Modified Roux-en-Y Gastric Bypass in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.zchf2t6 Copy   

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Authors: Jeff Hodgin
Group: Mouse Metabolic Phenotyping Centers
Summary: Immunoblotting (also called Western Blotting) is a technique to separate and identify individual proteins in a protein mixture (e.g. a cell lysate). In Immunoblotting (Western blotting) the protein mixture is applied to a gel electrophoresis (SDS-PAGE gel electrophoresis) to sort the proteins by its molecular weight size in individual protein bands. The separated protein bands are then transferred to a nitrocellulose transfer membrane. The proteins adhere to the membrane in the same pattern as they have been separated due to interactions of charges. To visualize the protein of interest the membrane is commonly first probed using a primary protein- specific antibody followed by a labeled secondary antibody. The specificity of the antibody-antigen interaction enables a target protein to be identified in the midst of a complex protein mixture. The bound antibodies are then detected by western blotting substrate (e.g. Pierce ECL Western Blotting Substrate), and then visualized by using adeveloping film (HyBlot CL Autoradiography Film) in the darkroom. The thickness of the band corresponds to the amount of protein present.

Proper citation: Jeff Hodgin 2019. U Michigan - Western Blot. protocols.io https://dx.doi.org/10.17504/protocols.io.56hg9b6 Copy   

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Authors: Lin Zhong, Jeffrey Rottman, Chee Lim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:The most common cause of cardiovascular mortality in man is the outcome from myocardial infarction. Accordingly, it is necesarry to study the corresponding process of heart injury recovery in many mouse models relevant to human cardiovascular disease. This protocol describes the surgical induction of myocardial injury via 1) permanent ligation of a coronary artery in the heart (myocardial infarction), 2) cryoinjury, and 3) transient occlusion of a coronary artery followed by reperfusion (ischemia-reperfusion injury).

Proper citation: Lin Zhong, Jeffrey Rottman, Chee Lim 2019. Vandy - Mouse Myocardial Infarction. protocols.io https://dx.doi.org/10.17504/protocols.io.6xghfjw Copy   

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Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: Cayman Chemical's Protein Carbonyl Colorimetric Assay Kit is a convenient colorimetric assay for the measurement of oxidized proteins. Protein samples are derivatized by making use of the reaction between 2,4-dinitrophenylhydrazine (DNPH) and protein carbonyls. Formation of a Schiff base produces the corresponding hydrazone which can be analyzed spectrophotometrically at 360-385 nm. This assay can be used to measure oxidized protein in plasma, serum, cell lysates, and tissue homogenates.

Proper citation: Peter Havel 2019. UC Davis - Protein Carbonyl. protocols.io https://dx.doi.org/10.17504/protocols.io.63hhgj6 Copy   

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Authors: John Stack, Gary Cline
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure used to measure the concentration of Blood Urea Nitrogen(BUN) in blood, plasma, and serum. Urea is determined by the enzymatically coupled reactions of urease (to form ammonia) and glutamate dehydrogenase (conversion of ammonia and glutamate to glutamine with oxidation of NADH to NAD). The rate of NAD formation is monitored by the change in absorbance at 340 nm.

Proper citation: John Stack, Gary Cline 2019. Yale - Blood Urea Nitrogen. protocols.io https://dx.doi.org/10.17504/protocols.io.y3qfymw Copy   

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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 https://dx.doi.org/10.17504/protocols.io.xxbfpin Copy   

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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 https://dx.doi.org/10.17504/protocols.io.x3hfqj6 Copy   

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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 https://dx.doi.org/10.17504/protocols.io.x5gfq3w Copy   

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