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HORMONES AND SIGNALING
1Center for Ulcer Research and Education, Center for Digestive Disease Research, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles 90073 and Digestive Diseases Division, University of California Los Angeles School of Medicine, Los Angeles 90024; 2Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229; and 3Division of Gastroenterology, Harbor University of Los Angeles Medical Center, Torrance, California 90509
Submitted 11 December 2002 ; accepted in final form 3 April 2003
CCK-58 differs from CCK-8 in patterns of expression of pancreatic secretion of fluid and amylase and gallbladder contraction. These differences have physiological relevance only if CCK-58 release is stimulated by nutrients entering the intestine and if CCK-58 circulates in sizeable amounts. In this study, we report that when radiolabeled CCK-58 is added to rat blood and plasma is formed, there is extensive loss and degradation of the radioactive peptide. Therefore, a new method was developed to minimize loss and degradation of this label. This method recovered >85% of the label with no detectable degradation. Furthermore, the optimized method recovered all unlabeled exogenous cholecystokinin molecular forms in >80% yields. Blood from fasted rats and rats in which cholecystokinin release was stimulated by the trypsin inhibitor camostat contained only CCK-58 (3.5 ± 0.5 and 17 ± 1.5 fmol/ml, respectively). Because CCK-58 predominates in the blood, this molecular form should be used in studies on the physiology and pathophysiology of cholecystokinin.
pancreas; radioimmunoassay; blood processing; plasma; molecular forms
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