Test descriptions


  • CA2000:  Body Composition
    Food consumption is measured over a 24-hour period. This is done once a week for the duration of the study.
    (Link to: www.mmpc.org Shared Catalog)
    (Link to related publication(s))



  • CA2001:  Food Consumption
    Food consumption is measured over a 24-hour period. This is done once a week for the duration of the study.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2002:  Body Weight
    Mice are weighed 1-3 times per week for the duration of the study.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2003:  Continuous measurement of body temperature (by probe)
    Body temperature will be measured by anal probe for a short period of time. This can be done at room temperature or under cold challenge conditions.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2004:  GTT - Glucose Tolerance Test
    Mice will be fasted for 5 hours. A fasting blood sample will be removed from the tail vein and a concentrated solution of glucose injected into the abdominal cavity of the mice through a needle passed through the abdominal skin. 

    Blood samples (~ 5ul) will be removed from the tail vein 15, 30, 60 and 120 minutes later and glucose will be measured by glucometer.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2005: Insulin concentrations at fasting and post intraperitoneal glucose administration
    Mice will be fasted for 5 hours. A fasting blood sample will be removed from the tail vein and a concentrated solution of glucose injected into the abdominal cavity of the mice through a needle passed through the abdominal skin.  Thirty minutes later, another blood sample will be taken.  Insulin in plasma at time 0 and 30 min. will be measured by ELISA. (CA2006)
    (Link to: www.mmpc.org Shared Catalog)



  • CA2006:  Plasma insulin measurement by ELISA
    We will measure insulin concentration in plasma from mice (~25ul/mouse in duplicate) by commercially available ELISA.   (Link to: www.mmpc.org Shared Catalog)



  • CA2007:  Insulin concentrations at fasting and post intraperitoneal insulin administration
    Insulin concentrations at fasting and post intraperitoneal insulin administration.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2008:  Insulin Tolerance Test (ITT)
    Glucose concentrations at fasting and post intraperitoneal insulin administration.

    Mice will be fasted for 5 hours and anesthetized.  A fasting blood sample will be removed from the tail vein and insulin (0.5mU/g) will be injected into the abdominal cavity. Blood samples will be removed from the tail vein 15, 30, 45 and 60 minutes later.  Samples of plasma obtained during the test will be measured for concentrations of glucose and insulin.  (Link to: www.mmpc.org Shared Catalog)



  • CA2009:  Triglycerides in liver
    A small piece of liver tissue will be homogenized and liver triglycerides will be saponified in KOH. Glycerol will be measured against glycerol standards using a commercially available reagent set.
    (Link to: www.mmpc.org Shared Catalog)


  • CA2010:  Plasma Triglycerides
    Plasma triglycerides will be saponified in KOH. Glycerol will be measured against glycerol standards using a commercially available reagent set.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2011:  Total Energy Expenditure using (2H-labeled water)
    TEE is equal to the sum of basal metabolic metabolic, thermic effect of eating and physical activity. Following a single bolus injection of 2H and 18O-labeled water one can determine TEE via the elimination of 2H and 18O from body water. This test requires serial measurements of the labeling of body water over approximately 1 week, which necessitates the collection of blood or urine samples.

    Note: This test does not require catheterized mice, nor does it require that mice be shipped to the MMPC. The isotopes are non-radioactive and no special safety precautions are required, the tracers will be shipped from the MMPC to the investigator. Investigators will be instructed on how to administer the isotopes, collect samples and then ship them back to the MMPC.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2013:  Hyperinsulinemic Clamp (Hypoglycemic or Euglycemic) using stable isotopes
    The euglycemic-hyperinsulinemic clamp is used to investigate insulin action and glucose metabolism in the intact, conscious mouse. The use of different glucose or glucose analog tracers with the option of using dual tracers (3-H or 14-C labeled) allows the determination of tissue-specific glucose uptake in addition to whole body glucose production and disposal. We can also use stable isotopes for the glucose clamp studies.

    A chronic in-dwelling catheter is surgically implanted into the right jugular vein of the mouse.  After a minimum of 4 days of recovery, the mice are fasted the morning of the experiment.  The 120-minute protocol begins with a prime-continuous infusion of glucose tracer and a prime-continuous infusion of insulin, at a rate dependant on the goals and conditions of the experiment.  Glucose is infused at a rate that maintains euglycemia and is a measure of insulin sensitivity.  Tissues can be collected at the conclusion of the clamp for later assessment of tissue-specific glucose uptake as well as other biochemical assays as desired.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2015: Turnover of glucose, lipid and/or protein
    During a constant tracer infusion, the dilution of the infused tracer yields a measure of that molecule's rate of appearance. One can measure the turnover of numerous molecules using this strategy. One can determine the kinetics glucose, glycerol and protein using [6,6-2H2]glucose, [2H5]glycerol and [2H5]phenylalanine.

    Note: This test requires a catheterized animal.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2016:  Fatty acid and cholesterol synthesis using 2H-labeled water
    Rates of fatty acid and cholesterol synthesis can be determined in tissues via the incorporation of 2H or 13C-labeled precursors. For example, following a bolus injection of 2H-labeled water one can collect samples (e.g. blood, liver and/or adipose tissue). The respective lipids are isolated and their 2H-labeling is determined. This test can be performed in 2 modes, short term vs long term.
    In a short term study, the tracer is administered and samples are collected within hours to determine the synthesis of lipids in plasma and/or liver. In a long term study, the tracer is continuously administered over several days.  Samples of adipose tissue are collected. The difference in time scale is necessary since the pool of lipids in adipose tissue is relatively large and requires more time for label to appear.

    Note: This test does not require catheterized mice, nor does it require that mice be shipped to the MMPC. The isotopes are non-radioactive and no special safety precautions are required, the tracers will be shipped from the MMPC to the investigator. Investigators will be instructed on how to administer the isotopes, collect samples and then ship them back to the MMPC.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2017:  Tissue-specific protein synthesis using 2H-labeled water
    Rates of protein synthesis can be determined from the incorporation of 2H-labeled water. For example, following a bolus injection of 2H-labeled water one can collect samples (e.g. blood, liver, muscle, etc). Total proteins are isolated and their 2H-labeling is determined. This test can be performed in 2 modes, short term vs long term. In a short term study, the tracer is administered and samples are collected within hours to determine the synthesis of proteins in plasma, liver, etc.  

    This mode is well-suited for examining the acute response of protein synthesis to a perturbation (e.g. food intake).  In a long-term study, the tracer is continuously administered over several days.  Samples are collected and the assays are performed. The long term design yields an integrative measure of protein synthesis;  i.e.: the isotope is present during the fed and the fasted state and accounts for all protein synthesis over such a transition.

    Note: This test does not require catheterized mice, nor does it require that mice be shipped to the MMPC. The isotopes are non-radioactive and no special safety precautions are required, the tracers will be shipped from the MMPC to the investigator. Investigators will be instructed on how to administer the isotopes, collect samples and then ship them back to the MMPC.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2018:  Profile of acylcarnitines in plasma/urine or tissue samples
    These LC-MS/MS assays are routinely run in which acylcarnitines are identified ac Cx, where x is the number of carbons in the acyl group. Samples are spiked with unlabeled and labeled internal standards. The mass isotopomer distribution of each peak is determined to characterize its labeling pattern.

    This test, coupled with the assay of the profile of urinary organic acids helps in the characterization of a number of metabolic defects, such as inborn errors of fatty acid oxidation disorders.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2019:  Profile of long chain acyl-CoAs in tissue
    Commercial preparations of CoA and acyl-CoA contain an unnatural analog of CoA, iso-CoA, in which the 3' phosphate has been moved to the 2' position of ribose. We can use the acyl-iso-CoA esters as internal standards to calculate the concentration and mass isotopomer distribution of acyl-CoAs from LC-MS data.
    (Link to List of Phenotype Assays CATALOG)



  • CA2020:  Measurement of acetyl-CoA, propionyl-CoA and/or succinyl-CoA in tissue
    Commercial preparations of CoA and acyl-CoA contain an unnatural analog of CoA, iso-CoA, in which the 3' phosphate has been moved to the 2' position of ribose. We can use the acyl-iso-CoA esters as internal standards to calculate the concentration and mass isotopomer distribution of acyl-CoAs from LC-MS data.   (See list of phenotype assays HERE)



  • CA2022:  13C-Labeling pattern of acetyl moiety of citrate (substrate oxidation)
    A number of investigators, who use 13C-labeled precursors of acetyl-CoA -in vivo- in isolated organs or in cell incubations, have attempted to estimate the labeling of mitochondrial acetyl-CoA to calculate the contribution of the substrate to the acetyl-CoA oxidized in the citric acid cycle (CAC). The best proxy for mitochondrial acetyl-CoA is the acetyl moiety of citrate.

    We developed an assay of the labeling of the acetyl moiety of citrate which involves (i) tissue extraction, (ii) alkaline hydrolysis of extant acetyl-CoA, (iii) after pH adjustment, cleavage of citrate with CoA + ATP-citrate lyase which we isolated from rat liver.

    The acetyl-CoA formed is either assayed as such by LC-MS, or reacted with thiophenol, followed by GC-MS assay of acetylthiophenol. This assay allows one to calculate the contribution of two or three substrates to mitochondrial acetyl-CoA in the same experiment.

    For example, consider a mouse heart perfused with unlabeled glucose + [1-13C]palmitate + [U-13C4]acetoacetate.

    These substrates yield acetyl-CoA that is unlabeled (M), singly labeled (M1), or doubly labeled (M2), respectively.  So the percent abundances of the M, M1, and M2 mass isotopomers of the acetyl moiety of citrate yield the contribution of each of the substrates to mitochondrial energy production.  (Link to: www.mmpc.org Shared Catalog)



  • CA2024:  Metabolomic profile of citric acid cycle and gluconeogenic intermediates
    We will assay the relative concentration of citric acid cycle intermediates and those in the gluconeognic pathway. Assays can be run using samples from mice/organs that have also been infused with a 13C-labeled tracer, e.g. 13C-lactate.  This strategy allows one to determine flux rates (via the 13C-labeling patterns) and identify points of control of a pathway, e.g. gluconeogenesis (via the relative concentration profiles).
    (link to mmpc.org shared catalog and more on catalog group: Carbohydrate Metabolism)



  • CA2024CT:  CUSTOM-DESIGNED TRACER EXPERIMENT
    We help users design experiments aimed at measuring: (i) carbon flux through a pathway, or  (ii) the distribution of carbon fluxes through multiple pathways.

    We emphasize the need to distinguish net carbon flux from isotopic flux. The latter often results from net carbon flux, isotopic exchange, or both. We suggest the use of isotopic substrate(s) and analytical equipment (mass spectrometry, NMR) most appropriate to the problem being studied. If the isotopic substrate is not commercially available, we can, in some cases, arrange for a pilot synthesis of the compound in an academic lab.

    We welcome users and their staff to spend time in our lab to learn and practice isotopic techniques.  (INQUIRE with Dr. Brunengraber  or  Dr. Puchowicz directly, if necessary)



  • CA2025:  Chronic arterial and jugular vein catherization
    Mice are anesthetized with isoflurane. Under sterile surgical conditions, permanent catheters are inserted into one jugular vein and one carotid artery. The catheters are funneled to the back of the neck, filled with heparinized saline and plugged. The skin is sutured. The mice are allowed to recover for 2-4 days before being used for metabolic studies if they have recovered their pre-surgical body weight. The patency of the catheters is periodically checked, by flushing with heparinized saline.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2026:  Chronic arterial or jugular vein catherization
    Mice are anesthetized with isoflurane. Under sterile surgical conditions, a permanent catheter is inserted into one jugular vein or one carotid artery. The catheter is funneled to the back of the neck, filled with heparinized saline and plugged. The skin is sutured. The mice are allowed to recover for 2-4 days before being used for metabolic studies if they have recovered their pre-surgical body weight. The patency of the catheter is periodically checked, by flushing with heparinized saline.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2027:  Acute arterial and jugular vein catherization
    Mice are anesthetized with isoflurane. Under sterile surgical conditions, permanent catheters are inserted into one jugular vein and one carotid artery. After 30 min of recovery, during which rectal temperature is monitored and maintained at 37ºC, the catheters are connected to syringe pumps or syringes and the acute metabolic investigation is started. At the end of the experiment, the mouse is euthanized by a bolus injection of pentobarbital.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2028:  Acute arterial or jugular vein catherization
    Mice are anesthetized with isoflurane. Under sterile surgical conditions, a permanent catheter is inserted into one jugular vein or one carotid artery. After 30 min of recovery, during which rectal temperature is monitored and maintained at 37ºC, the catheter is connected to a syringe pump or a syringe and the acute metabolic investigation is started. At the end of the experiment, the mouse is euthanized by a bolus injection of pentobarbital.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2029:  Acute portal vein catherization
    Mice are anesthetized with isoflurane. Under sterile surgical conditions, a permanent catheter is inserted into the portal vein through mid-line laparotomy. The skin is sutured. After 30 min of recovery, during which rectal temperature is monitored and maintained at 37ºC, the catheter is connected to a syringe pump or a syringe and the acute metabolic investigation is started. At the end of the experiment, the mouse is euthanized by a bolus injection of pentobarbital.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2030:  Implant [G2 E – Mitters™]
    Implant [G2 E – Mitters™].
    (See list of Metabolite Concentration and Enrichment HERE)



  • CA2031:  Long-term analysis of surgery implantation on Min-Mitter™
    Long-term analysis of surgery implantation on Min-Mitter™.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2040:  Metabolomic profile of Free Fatty Acids/sterols in Plasma, Urine or Tissue
    Metabolomic profile of free fatty acids in tissue.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2041:  Tissue processing by Pathology Core
    Tissue processing by Pathology Core (embedded in paraffin)
    (Link to: www.mmpc.org Shared Catalog; ref. publication)



  • CA2043:  Portal vein injection
    To observe the effects in insulin signaling, insulin (1 milli-unit · kg-1 body weight) will be administered to anesthetize mice by injection into the portal vein. Liver, skeletal muscle and fat biopsies will be removed before basal-time (0) and 5 min after the insulin injection. The tissues will be quick-frozen for the investigator's future analysis, using the Western Blot Procedure.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2044:  Brain uptake and blood flow
    Brain uptake and blood flow.
    (Link to: www.mmpc.org Shared Catalog)



  • CA2045:  Measurement of ATP/ADP
    ATP tissue concentrations will be analyzed enzymatically from 25-30 mg of tissue and reported as nmol/mg wet tissue weight.
    (Link to: www.mmpc.org Shared Catalog and the Metabolite Concentration and Enrichment Catalog Group)



  • CA2046:  Indirect Calorimetry / First 36-hr measurement (for 8 mice)
    Both total energy expenditure and relative rates of carbohydrate versus fat oxidation will be determined via indirect calorimetry measured by CO2 production in specially-designed chambers for mice.
    (Link to: www.mmpc.org Shared Catalog and the 'Energetics' and 'Energy Expenditure & Exercise' Catalog Groups)



  • CA2047:  Treadmill Training/Endurance Study PLUS Indirect Calorimetry (for 8 mice)
    Both total energy expenditure and relative rates of carbohydrate versus fat oxidation will be determined via indirect calorimetry measured by CO2 production in specially designed treadmill chambers for mice.
    (Links to related www.mmpc.org Energy Expenditure & Exercise CATALOG; and Metabolite Concentration and Enrichment CATALOG)



  • CA2048:  Whole Body Fixation (PFA) + Tissue Collection
    Both total energy expenditure and relative rates of carbohydrate versus fat oxidation will be determined via indirect calorimetry measured by CO2 production in specially designed treadmill chambers for mice.
    (Links to related www.mmpc.org Imaging CATALOG; and Isolated Organ and Cell Perfusion CATALOG and Pathology & Immunohistochemistry CATALOG)



  • CA2049:  Additional 24-hr measurement (for 8 mice)
    Additional 24-hr measurement (for 8 mice).
    (Links to www.mmpc.org Energy Expenditure & Exercise CATALOG; and Metabolite Concentration and Enrichment CATALOG)



  • CA2050:  Data Summary Interpretation
    Data Summary Interpretation.
    (Links to www.mmpc.org Shared Catalog; and related Miscellaneous Tests CATALOG)



  • CA2051:  Excise Tissues, Blood Serum/Plasma
    Excise Tissues, Blood Serum/Plasma.
    (Links to www.mmpc.org Shared Catalog; and Pathology & Immunohistochemistry CATALOG)



  • CA2052:  Isolated mouse HEART perfusion
    Isolated mouse HEART perfusion.
    (Links to www.mmpc.org Shared Catalog; and the Isolated Organ and Cell Perfusion CATALOG)



  • CA2053:  Isolated mouse LIVER perfusion
    Isolated mouse LIVER perfusion.
    (Links to www.mmpc.org Shared Catalog; the Isolated Organ and Cell Perfusion CATALOG and the Liver Function CATALOG)



  • CA2053CB:  CUSTOM-DESIGNED BIOLOGICAL EXPERIMENT
    We help users to design and implement in vivo experiments aimed at unraveling the metabolic process they are investigating. Protocols use a combination of physiological, metabolic and, if needed, isotopic techniques. The experiments and/or analyses are conducted in the user's lab or at the MMPC, with some of the analyses conducted in one of the MMPCs.
    (Links to www.mmpc.org Shared Catalog; and related Miscellaneous Tests CATALOG)



  • CA2055:  Quarantine (4 weeks) mice imported to Case
    Quarantine (4 weeks) mice imported to Case Western Reserve University.
    (Links to www.mmpc.org Shared Catalog; and related Miscellaneous Tests CATALOG)



  • CA2056:  Housing mice (1 to 14 days)
    Housing mice (1 to 14 days).
    (Links to www.mmpc.org Shared Catalog; and related Miscellaneous Tests CATALOG)