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Insulin is secreted primarily in response to glucose, while other nutrients such as free fatty acids and amino acids can augment glucose-induced insulin secretion. In addition, various hormones, such as melatonin, estrogen, leptin, growth hormone, and glucagon like peptide-1 also regulate insulin secretion.
Insulin sensitivity is calculated as the glucose infusion rate during the second hour (M) divided by the mean insulin concentration at 60, 90, and 120 min (I), and the clamp glucose clearance per unit of insulin is calculated as M/I divided by the clamped glucose concentration.
The glucose clamp method is the reference standard for direct measurement of insulin sensitivity. Regarding simple surrogates, QUICKI and Log (HOMA) are among the best and most extensively validated. Dynamic tests are useful if information about both insulin secretion and insulin action are needed.
Basic Considerations Pre fasting for 6 or 12 hrs. Measure mice body weight after fasting. Calculate the amount of glucose and insulin needed. Cut tail and measure basal glucose in the blood. i.p. with a time-lapse between mice of 3 min. Repeat determination of blood glucose concentration after 15, 30, 60, 90 and 120 min.
Increased delivery of insulin into the circulation causes further suppression of hepatic glucose release (to 0.5 mg/kg/min) and increased stimulation of glucose uptake by insulin-sensitive tissues such as muscle to restore normoglycemia.
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In mice, the GTT is the simplest, and usually the first, test applied to an animal model, and it provides a physiological overview of any changes in glucose tolerance without determining the causative mechanisms. Briefly, mice are routinely fasted before the administration of glucose.
Hyperinsulinemic-euglycemic clamp: The gold standard for evaluating insulin sensitivity, this clamp technique requires a steady IV infusion of insulin to be administered in one arm. The serum glucose level is clamped at a normal fasting concentration by administering a variable IV glucose infusion in the other arm.

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