Following the basal glucose turnover study, the same cohort of ob/ob (B6) mice was subjected to an FSIGT. Blood sampling was performed via an arterial catheter in unrestrained, conscious animals. A continuous infusion of saline-washed erythrocytes was commenced at t = 0 min to prevent a >5% fall in hematocrit. Baseline fasting blood samples were drawn at −10 and 0 min. Based on a published protocol22 (link), a bolus of 50% dextrose (0.75 g/kg body weight) was injected iv over a period of 15s at t = 0 min. Blood (20 μl) was sampled both for measurement of glucose using a hand-held glucometer (Accu-Chek Aviva Plus, Roche; Indianapolis, IN ) and for subsequent assay of plasma insulin and lactate levels at time points 1, 2, 4, 8, 12, 16, 20, 30 and 60 min after the glucose injection. Additional samples were obtained for blood glucose measurement at 3, 5, 6, 10, 14, 18, 25, 40 and 50 min using a hand-held glucometer.
Protocol detail
Intravenous Glucose Tolerance Test in ob/ob Mice
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Animals
Arterial
Assay
Blood
Blood glucose
Body weight
Catheter
Conscious
Erythrocytes
Glucose
Held
Hematocrit
Insulin
Lactate
Mice
Plasma
Saline
Corresponding Organization :
Other organizations : University of Washington, University of Pennsylvania, Cedars-Sinai Medical Center, Vanderbilt University, St. Joseph's Hospital and Medical Center, Barrow Neurological Institute, University of Illinois at Chicago
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Variable analysis
independent variables
- Glucose injection: A bolus of 50% dextrose (0.75 g/kg body weight) was injected iv over a period of 15s at t = 0 min.
- Blood glucose levels measured using a hand-held glucometer at time points 1, 2, 4, 8, 12, 16, 20, 30 and 60 min after the glucose injection, and additional measurements at 3, 5, 6, 10, 14, 18, 25, 40 and 50 min.
- Plasma insulin levels measured at time points 1, 2, 4, 8, 12, 16, 20, 30 and 60 min after the glucose injection.
- Plasma lactate levels measured at time points 1, 2, 4, 8, 12, 16, 20, 30 and 60 min after the glucose injection.
- Continuous infusion of saline-washed erythrocytes commenced at t = 0 min to prevent a >5% fall in hematocrit.
- Baseline fasting blood samples were drawn at −10 and 0 min.
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