Copper deficiency increases total protein and apolipoprotein A-I synthesis in the rat small intestine.
This study was designed to determine whether an enhanced intestinal synthesis of apolipoprotein (apo) A-I is associated with the hyperapolipoproteinemia observed in copper-deficient rats. Male weanling Sprague-Dawley rats were assigned to two dietary treatments, Cu deficient (0.6 ppm Cu) and Cu adequate (6.0 ppm Cu) for 6 weeks. In vivo studies were then performed after rats were injected with a flooding dose of 150 microM [3H]phenylalanine (PHE, 50 microCi/ml/100 g body wt). Three rats from each treatment were sacrificed at 5, 10, 15, 30, and 60 min postinjection. The small intestine was rapidly rinsed and frozen in liquid N2. In vitro studies were performed by labeling freshly isolated 6-cm segments from duodenum, jejunum and ileum with [3H]PHE (33 microCi/ml, 49.7 Cl/mmol) in PHE-free minimum essential medium for 7 and 14 min. In vivo and in vitro intestinal samples were sonicated, solubilized in 1% Triton X-100, and centrifuged to provide the detergent soluble fraction for the isolation of nascent apo A-I and total protein. Radioactivities associated with nascent apo A-I isolated by immunoprecipitation and SDS-PAGE, and with total protein precipitated by trichloroacetic acid, were measured to determine the influence of Cu deficiency on nascent apo A-I and total protein synthesis. In the Cu-deficient small intestine, the synthesis of total protein was measured only in the duodenum and was enhanced after 1 hr for the in vivo studies. Moreover, total protein synthesis was enhanced at both 7 and 14 min of the in vitro studies for all three small intestinal segments of the Cu-deficient rats. Apo A-I synthesis was measured only at the jejunum and was also enhanced by Cu deficiency in the in vitro studies. Thus, an increase in intestinal apo A-I synthesis may contribute to the elevated plasma apo A-I level in Cu-deficient rats.
Wu
J Y
JY
Department of Nutritional Sciences, University of Arizona, Tucson 85721, USA.
Lei
K Y
KY
D000818:Animals; D016632:Apolipoprotein A-I / Q000096:biosynthesis*; D003300:Copper / Q000172:deficiency*; D004032:Diet; D004195:Disease Models, Animal; D006951:Hyperlipoproteinemias / Q000209:etiology / Q000378:metabolism; D066298:In Vitro Techniques; D007421:Intestine, Small / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D014176:Protein Biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley
D016632:Apolipoprotein A-I; D003300:Copper