The application of 2-NBDG as a fluorescent tracer for assessing hepatic glucose production in mice during hyperinsulinemic euglycemic clamp

摘要

Methods of performing insulin clamps vary between laboratories. Here we present a protocol of insulin clamping in conscious mice, with the significant advantage of avoiding multiple surgical catheterizations and non-physiologic metabolism during the induction of anesthesia. Using this technique we also established a new method for measuring hepatic glucose production (HGP) using a fluorescent d -glucose analog, 2-[ N -(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxyglucose (2-NBDG). To prove the reliability and feasibility of this method, whole-body insulin sensitivity was compared between conscious normal ICR mice and diabetic KKAy mice using the insulin clamp. Basal and clamp HGP was compared between normal C57 mice and diabetic db/db mice by using the modified clamp with 2-NBDG as a tracer. The glucose infusion rate (GIR), an index of insulin sensitivity, was significantly lower in KKAy mice than normal ICR mice (6.2±1.3?mg/kg/min vs. 31.3±2.9?mg/kg/min, P Graphical We present a protocol of insulin clamping in conscious mice, with the significant advantage of avoiding multiple surgical catheterizations and non-physiologic metabolism during the induction of anesthesia. Figure options