Animal Characterization Core

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Animal Characterization Core
University of Virginia DERC

• Dr. Susanna Keller, Director
• Melissa Horal, Lab Specialist
• Toni Barbera, Lab Specialist

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Animal Characterization Core

• Physiology Laboratory
• Provides facilities to measure metabolic and
  hemodynamic parameters in vivo in rats and mice
• Facilitates access to MRI, ultrasound and DEXA,
  all instruments for in vivo assessment of
  anatomic and functional abnormalities in
• rats and mice

• Assay Laboratory
• Conducts routine assays to determine circulating
  levels of hormones, metabolites and intracellular
  signaling molecules
• Provides access to specialized equipment
• Provides advice on evaluating hormone action at
  the cellular level

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Assay Laboratory
Located in Aurbach Medical Research Building,
Fontaine Research Park, Room 1255 Staffed by
Toni Barbera, Lab Specialist
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Services Provided
Assays Currently established Insulin (Human),
C-peptide (Human), glucagon, cAMP and cGMP,
hemoglobin A1c, glucose To be established
Insulin (Rat and Mouse), Free Fatty Acids, others
upon request and demand
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Access to Equipment
Molecular Dynamics Typhoon Equipped with
scanning lasers for two dimensional detection of
b- and g-emitting radioactive materials (e.g. 32P
and 125I) fluorescent chromophores, and
light-emitting particles Applications
Scanning of protein and DNA gels, labeled
peptides or amino acids on thin layer
chromatography plates Quantitation of the signals
through ImageQuant software PC connected to the
network for easy file transfer. The instrument
is located on the ground floor of the Aurbach
Medical Research Building
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Access to Equipment

• HPLC System
• Beckman HPLC instrument connected to a System
  Gold operating system unit
• ELISA Equipment
• a Flo Instrument microtiter plate washer and a
  Titertek plate reader.
• Radioisotope Counters
• LKB four-channel gamma-counter and a liquid
  scintillation counter.
• All of the above are located in the Assay
  Laboratory

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Other Services Advice on standard techniques
for analysis of insulin signaling at the
cellular level Development of New Techniques
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Physiology Laboratory

• Located in Animal Facility in Aurbach Medical
  Research Building, Fontaine Research Park,
• Room G-131
• Staffed by Melissa Horal, Lab Specialist

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Services Provided

• Collection of blood samples for the determination
  of blood parameters
• (peptide hormone levels, blood metabolites)
• Insulin and Glucose Tolerance Tests
• For rats and mice

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Insulin and Glucose Tolerance Tests
Insulin tolerance test Animals will be fasted
for 6 hours before the experiment. Insulin (0.75
U/kg) in 0.1 ml 0.9 NaCl will be injected
intraperitoneally into non-anesthetized mice. A
drop of blood will be taken from the nicked tail
vein before the injection of insulin and after
10, 20, 30, 45, and 60 min for the determination
of blood glucose with a glucometer. Glucose
tolerance test The procedure is the same as for
the insulin tolerance test, but instead of
insulin, glucose is injected intraperitoneally (2g
/kg) and the mice are fasted overnight (16 hours)
before the test. Blood samples will be taken
before and at 10, 20, 30, 60, 90 and 120
min after the injection of glucose. The same
mice can be used for both tests. After the first
test the mice are allowed to recover for two
weeks before the second test is performed. A
minimum of eight mice per group will be used for
the tests
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Glucose and Insulin Tolerance Tests
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Services Provided

• Metabolic caging for automated non-invasive 24
  hour
• monitoring of food and water intake, urine
• excretion, activity levels and
• oxygen consumption and carbon dioxide production
• (up to 4 mice simultaneously)

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Comprehensive Laboratory Animal Monitoring System
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Metabolic Component
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Services Provided
Non-invasive blood pressure monitoring systolic,
diastolic blood pressure and heart rate (up to
4 rats or 4 mice simultaneously)
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Non-Invasive BP Monitor
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Future Services

• Additional metabolic caging
• (4 rats and an additional 4 mice)
• Expand BP monitor to accommodate
• (up to 8 mice or 8 rats simultaneously)
• Clamp studies
• Fat cell and muscle isolation and measurement of
  glucose uptake

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Ready to start the first experiments Blood
sampling, insulin and glucose tolerance tests
Beginning of February Metabolic cages and blood
pressure monitoring Beginning of
April Webpage with description of services
will be up at the beginning of February (will
include order forms in pdf format) Animal
Protocols Basic procedures are covered under
core protocols, approval for special procedures
will have to be obtained prior to the start of
experiments