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Publication
Considerations in the design of hyperinsulinemic-euglycemic clamps in the
conscious mouse.
Authors Ayala JE, Bracy DP, McGuinness OP, Wasserman DH
Submitted By David Wasserman on 11/5/2007
Status Published
Journal Diabetes
Year 2006
Date Published 2/1/2006
Volume : Pages 55 : 390 - 397
PubMed Reference 16443772
Abstract Despite increased use of the hyperinsulinemic-euglycemic clamp to study insulin
action in mice, the effects of experimental parameters on the results obtained
have not been addressed. In our studies, we determined the influences of
sampling sites, fasting duration, and insulin delivery on results obtained from
clamps in conscious mice. Carotid artery and jugular vein catheters were
implanted in C57BL/6J mice (n = 6-10/group) fed a normal diet for sampling and
infusions. After a 5-day recovery period, mice underwent a 120-min clamp (2.5-mU
. kg(-1) . min(-1) insulin infusion; approximately 120-130 mg/dl glucose) while
receiving [3-(3)H]glucose to determine glucose appearance (endoR(a)) and
disappearance (R(d)). Sampling large volumes (approximately 100 mul) from the
cut tail resulted in elevated catecholamines and basal glucose compared with
artery sampling. Catecholamines were not elevated when taking small samples (
approximately 5 mul) from the cut tail. Overnight (18-h) fasting resulted in
greater loss of total body, lean, and fat masses and hepatic glycogen but
resulted in enhanced insulin sensitivity compared with 5-h fasting. Compared
with a 16-mU/kg insulin prime, a 300-mU/kg prime resulted in hepatic insulin
resistance and slower acquisition of steady-state glucose infusion rates (GIR)
after a 5-h fast. The steady-state GIR was expedited after the 300-mU/kg prime
in 18-h-fasted mice. The GIR and R(d) rose with increasing insulin infusions
(0.8, 2.5, 4, and 20 mU . kg(-1) . min(-1)), but endoR(a) was fully suppressed
with doses higher than 0.8 mU . kg(-1) . min(-1). Thus, common variations in
experimental factors yield different results and should be considered in
designing and interpreting clamps.




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