Insulin Tolerance Test under Anaesthesia to Measure Tissue-specific Insulin-stimulated Glucose Disposal

Accurately assessing tissue-specific insulin action is essential for understanding disease progression and developing novel antidiabetic therapies.

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This protocol describes an insulin tolerance test (ITT) under anaesthesia in mice, combined with intravenous administration of the [³H]2-deoxyglucose (2-DOG) tracer. It enables quantitative assessment of insulin-stimulated glucose uptake specifically in adipose and muscle tissues, offering greater precision than conventional systemic ITT approaches.

Key Features

Tissue-Specific Readouts: Directly quantifies insulin-stimulated glucose disposal in adipose and skeletal muscle, moving beyond blood glucose-based endpoints.
Dynamic Tracer Kinetics: Measures [³H]2-DOG disappearance from the blood and accumulation of phosphorylated [³H]2-DOG-6-phosphate in tissues to calculate a glucose uptake index.
Pathway Validation: Allows downstream signaling analysis (e.g., Akt phosphorylation at Thr308 and Ser473) by Western blot to confirm insulin action at the molecular level.
Validated Model Applicability: Successfully applied in high-fat/high-sucrose diet-induced insulin-resistant mice, detecting impaired insulin-stimulated glucose uptake in epididymal adipose and quadricep muscles.

Applications

In vivo efficacy evaluation of antidiabetic drug candidates
Mechanistic studies of insulin resistance
Tissue-specific metabolic profiling

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Insulin Tolerance Test under Anaesthesia to Measure Tissue-specific Insulin-stimulated Glucose Disposal

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