ACURF Amendment Guidelines for the Addition of Glucose/Insulin Tolerance Test
(GTT/ITT)
January, 2014
The purpose of this document is to streamline the submission of amendments to IACUC protocols at the University of Iowa, specifically to add the use of the new Fraternal Order of Eagles / University of Iowa Diabetes Research
Center (FOE/DRC) Glucose/Insulin Tolerance Test (GTT/ITT).
Users should simply copy-and-paste some of the preformatted sections (below) into their existing IACUC protocols to satisfy the requirements of the University of Iowa IACUC.
Specific sections that should be copied verbatim include:

Question 4 (Personnel)

Question 10 (Special Housing)

Question 16 (Euthanasia)

Question 20 & 21 (antemortem blood samples)

Question 24 (food restriction)

Question 27 (Procedures)

Question 4 (Personnel):
Add the following personnel to protocol:
Name:
Address
Dept:
:
Phone #:
E-mail address:
Describe Training:
Describe exposure to each species that will occur during
Jamie Soto
140 EMRB
Internal Medicine
335-7962 jamie-soto@uiowa.edu
Extensive animal husbandry experience and familiar with all procedures on this protocol for which she is responsible.
X Yes Has this person taken the University online training module on
Policies/Regulations?
Describe related experience with techniques Proposed:
No
Experienced with all aspects of mouse work performed in the laboratory for which she is responsible. Over 7 years experience with Dr. Abel and 1- year prior.
Jamie is the technical director of the University of
Iowa/Fraternal Order of the Eagles’ Diabetes
Research Center Metabolic Phenotyping Core
Facility. She is the primary coordinator of the maintenance and use of the CLAMS unit, and has been involved in this unit’s operation since
November 2013. Jamie has received training directly from Columbus Instruments in the use of the CLAMS system, as well as from Justin Grobe who is the primary coordinator of the acquisition, installation and deployment of the CLAMS unit.
She is involved in assisting in euglycemic hyperinsulinemic glucose clamps for the Core
Facility. She also performs GTTs/ITTs for the Core.
N/A Describe how individual will be trained if no prior relevant training or experience
Is this person performing surgery?
X Survival Surgery
Nonsurvival surgery
Not Applicable
Exposure per day is highly variable based upon the procedures involved, ranging from a few minutes

performance of this proposal. per day, to 4 hours per day. Maximum of 20 hours / week

Question 10 (special housing):
Add the following special housing details to question 10:
10. Will any special housing conditions be required for this project (lighting, feed, special caging, biological containment, etc.)?
No; Proceed to Next Question
X Yes
Please Specify
Mice will be singly housed during GTT’s and ITT’s for the 6 hour fast and 3 hour procedure. This is to prevent fighting and to facilitate the ease of the nature of the experiment. They will be returned to normal housing at the end of the experiment.

Question 20 & 21 (antemortem blood samples):
Add the following antemortem blood samples details to question 20 & 21:
20. Is the “only” procedure performed the harvest of tissue and/or body fluids under anesthesia followed by euthanasia?
X No; Proceed to Question 21.
Yes; You are finished; you do not need to complete the rest of the form
21. Will antemortem blood samples be taken?
No; Proceed to Question 22.
X Yes
Species
Route
Mouse
Tail bleed
Amount
# of collections
5µL & 90µL
5µL: Up to 6 during a glucose/insulin tolerance test. 90µL: twice during a glucose/insulin tolerance test
5 – 30 minutes Interval between collections
Will blood collection be performed under anesthesia?
Yes;
X No
Anesthesia should be described in answer to Question 19.
Please copy and paste additional tables if needed.

Question 24 (food restriction):
Add the following food restriction details to question 24:
No ; Proceed to Question 25.
X Yes What is restricted? X Food Water
Species
Length of restriction
Interval between restrictions
Monitoring during restriction
Mouse
6 hours
1 week
Only during 3 hours of testing
Justify restricting food and/or water To normalize blood glucose levels before performing Glucose and Insulin Tolerance Tests as described in Q. #27, below.

Question 27 (Procedures):
Add the following (and customize) procedural details to question 27:
27. For each distinct procedure, describe the manipulations that will be performed (including surgery) and the order in which they will be performed. Include time frames and intervals for the sequence of procedures. Unless listed elsewhere in this document, describe all drugs
(including test substances) administered during the study as well as the dosage, route, and frequency. Procedures that will be performed after the animal has been euthanized need not be described. (Describe the procedures in enough detail so that reviewers will be able to determine what is actually being done to the animal. You DO NOT need to re-describe in detail procedures addressed in previous Questions (e.g anesthesia, euthanasia).
Glucose and Insulin Tolerance Tests:
Animals will be singly-housed for the duration of the fast (see Q. #24 above) and the 3 hour test, with no access to food. Samples will be collected from the mouse tail following a needle stick or scalpel nick into the lateral tail vein. Small samples (single drops) of blood will be collected at the same spot on the tail at baseline and after 5, 15, 30, 60, 90, and 120 minutes following delivery of glucose (1mg/kg in a 20% D-glucose formulation by IP injection), or insulin (0.5 to 2 units per kg body mass, IP). The maximum value given of each substance will be 10 milliliters per kilogram (10 microliters per gram) of body mass.
Thus, a 30 gram mouse receives 300 microliters (0.3 mL).
Because the volume required at each time point is approximately 5 microliters, the total blood volume collected over the entire test will not exceed the mandated maximum collection limits (test = less than 50 microliters, vs. limit = 1% of body mass, which for a
25g mouse would be 250 microliters). We also collect ~90microliters blood at T0 and T30 for serum to assess insulin levels. At least one week recovery will be allowed between serial tests on a single animal so as to clearly avoid weekly blood collection limits.
At the end of this procedure, animals will be returned to normal housing conditions, or sacrificed for tissue collection.