Considerations in the Design of Hyperinsulinemic-Euglycemic Clamps in the Conscious Mouse

• DOI: 10.2337/diabetes.55.02.06.db05-0686
• Publication Date: 2006-06-01T23:03:17Z
• Authors: Julio E. Ayala; Deanna P. Bracy; Owen P. McGuinness; David H. Wasserman
• Keywords: Basal (medicine); Jugular vein; Clamp; Blood sampling; Glucose clamp technique

Despite increased use of the hyperinsulinemic-euglycemic clamp to study insulin action in mice, effects experimental parameters on results obtained have not been addressed. In our studies, we determined influences sampling sites, fasting duration, and delivery from clamps conscious mice. Carotid artery jugular vein catheters were implanted C57BL/6J mice (n = 6–10/group) fed a normal diet for infusions. After 5-day recovery period, underwent 120-min (2.5-mU · kg−1 min−1 infusion; ∼120–130 mg/dl glucose) while receiving [3-3H]glucose determine glucose appearance (endoRa) disappearance (Rd). Sampling large volumes (∼100 μl) cut tail resulted elevated catecholamines basal compared with sampling. Catecholamines when taking small samples (∼ 5 tail. Overnight (18-h) greater loss total body, lean, fat masses hepatic glycogen but enhanced sensitivity 5-h fasting. Compared 16-mU/kg prime, 300-mU/kg prime resistance slower acquisition steady-state infusion rates (GIR) after fast. The GIR was expedited 18-h–fasted Rd rose increasing infusions (0.8, 2.5, 4, 20 mU min−1), endoRa fully suppressed doses higher than 0.8 min−1. Thus, common variations factors yield different should be considered designing interpreting clamps.