Copper (Cu) deficiency in rats reduces the relative concentration of duodenal hephaestin (Hp), reduces Iron (Fe) absorption, and causes anemia. An experiment was conducted to determine whether these effects could be reversed by dietary Cu repletion. Five groups of eight weanling male rats each were used. Group 1 was fed a Cu-adequate diet (5.0 mg Cu/kg; CuA) and Group 2 was fed a Cu-deflcient diet (0.25 mg Cu/kg; CuD) for 28 days. The rats were fed 1.0 g each of their respective diets labeled with 59Fe (37 kBq/g), and the amount of label retained was measured one week later by whole-body-counting (WBC). Group 3 was fed a CuA diet and Groups 4 and 5 were fed a CuD diet for 28 days. Group 5 was then fed the CuA dlet for another week while Groups 3 and 4 continued on their previous regimens. Rats In Groups 3, 4, and 5 were fed 1.0 g of diet labeled with 59Fe, and the amount of label retained was measured by WBC one week later. Rats were killed and duodenal enterocytes Isolated for Hp protein analysis, whole blood was analyzed for hematological parameters, and various organs for 59Fe content. CuD rats absorbed less (P <0.05) Fe than CuA rats, the relative amount of duodenal Hp was less (P <0.05) In CuD rate, and the CuD rats developed anemia. After the CuD rats had been repleted with Cu for one week, Fe retention rose to values even higher (P <0.05) than those in CuA rats. After two weeks, the relative amount of duodenal Hp was higher (P <0.05) than normal, and most signs of anemia were reversed. Liver 59Fe was elevated In CuD rats, but was restored to normal upon Cu repletion. These findings suggest a strong association between duodenal Hp abundance and Fe absorption In the CuD rat, and that reduced Fe absorption Is an important factor in the cause of anemia.
FoxPL. The copper-iron chronicles: the story of an intimate relationship. Biometals16: 9–40, 2003.
2.
JohnsonWT, SaariJT. Temporal changes in heart size, hematocrit and erythrocyte membrane protein in copper-deficient rats. Nutr Res11: 1403–1414, 1991.
3.
ReevesPG, DeMarsLCS. Copper deficiency reduces iron absorption and biological half-life in male rats. J Nutr134: 1953–1957, 2004.
4.
ReevesPG, RalstonNVC, IdsoJP, LukaskiHC. Contrasting and cooperative effects of copper and iron deficiencies in male rats fed different concentrations of manganese and different sources of sulfur amino acids in an AIN-93G-based diet. J Nutr134: 416–425, 2004.
5.
ZidarBL, ShadduckRK, ZeiglerZ, WinkelsteinA. Observations on the anemia and neutropenia of human copper deficiency. Am J Hematol3: 177–185, 1977.
ReevesPG, DeMarsLCS, JohnsonWT, LukaskiHC. Dietary copper deficiency reduces iron absorption and duodenal enterocyte hephaestin Protein in male and female rats. J Nutr135: 92–97, 2005.
VulpeCD, KuoYM, MurphyTL, CowleyL, AskwithC, LibinaN, GitschierJ, AndersonGJ. Hephaestin, a ceruloplasmin homologue implicated in intestinal iron transport, is defective in the sla mouse. Nat Genet21: 195–199, 1999.
10.
KuoYM, SuT, ChenH, AttiehZK, SyedBA, McKieAT, AndersonGJ, GitschierJ, VulpeCD. Mislocalisation of hephaestin, a multicopper ferroxidase involved in basolateral intestinal iron transport, in the sex linked anaemia mouse. Gut53: 201–206, 2004.
11.
NittisT, GitlinJD. Role of copper in the proteosome-mediated degradation of the multicopper oxidase hephaestin. J Biol Chem279: 25696–25702, 2004.
12.
McKieAT, MarcianiP, RolfsA, BrennanK, WehrK. Barrow D, Miret S, Bomford A, Peters TJ, Farzaneh F, Hediger MA, Hentze MW, Simpson RJ. A novel duodenal iron-regulated transporter, IREG1, implicated in the basolateral transfer of iron to the circulation. Mol Cell5: 299–309, 2000.
13.
ZollerH, KochRO, TheurlI, ObristP, PletrangeloA, MontosiG, HaileDJ, VogelW, WeissG. Expression of the duodenal iron transporters divalent-metal transporter 1 and ferroportin 1 in iron deficiency and iron overload. Gastroenterology120: 1412–1419, 2001.
14.
EvansJL, AbrahamPA. Anemia, iron storage and ceruloplasmin in copper nutrition in the growing rat. J Nutr103: 196–201, 1973.
15.
National Research Council. Guide for the Care and Use of Laboratory Animals. Washington, DC: National Academy Press, 1996.
16.
ReevesPG, NielsenFH, FaheyGCJr.AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc committee on the reformulation of the AIN-76A rodent diet. J Nutr123: 1939–1951, 1993.
17.
ReevesPG. AIN-93 purified diets for the study of trace element metabolism in rodents. In: WatsonRD, Ed. Trace Elements in Laboratory Rodents. Boca Raton: CRC Press, pp 3–37, 1996.
18.
SchosinskyKH, LehmannHP, BeelerMF. Measurement of ceruloplasmin from its oxidase activity in serum by the use of o-dianisidine hydrochloride. Clin Chem20: 1556–1563, 1974.
19.
CrapoJD, McCordJM, FridovichI. Preparation and assay of superoxide dismutases. Methods Enzymol53: 382–393, 1978.
20.
FattmanCL, EnghildJJ, CrapoJD, SchaeferLM, ValnickovaZ, OuryTD. Purification and characterization of extracellular superoxide dismutase in mouse lung. Biochem Biophys Res Commun275: 542–548, 2000.
21.
ChenH, AttiehZK, SuT, SyedBA, GaoH, AlaeddineRM, FoxTC, UstaJ, NaylorCE, EvansRW, McKieAT, AndersonGJ, VulpeCD. Hephaestin is a ferroxidase that maintains partial activity in sex linked anemia mice. Blood103: 3933–3939, 2004.
22.
GublerCJ, LaheyME, ChaseMS, CartwrightGE, WintrobeMM. Studies on copper metabolism, HI: the metabolism of iron in copper deficient swine. Blood7: 1075–1092, 1952.
23.
LeeGR, NachtS, LukensJN, CartwrightGE. Iron metabolism in copper-deficient swine. J Clin Invest47: 2058–2068, 1968.
24.
CoppenDE, DaviesNT. Studies on the roles of apotransferrin and caeruloplasmin (EC 1.16.3.1) on iron absorption in copper-deficient rats using an isolated vascularly- and luminallv-perfused intestinal preparation. Br J Nutr60: 361–373, 1988.
25.
ThomasC, OatesPS. Copper deficiency increases iron absorption in the rat. Am J Physiol Gastrointest Liver Physiol285: G789–G795, 2003.
26.
DancisA, YuanDS, HaileD, Ask withC, EideDJ, MoehleC, KaplanJ, KlausnerRD. Molecular characterization of a copper transport protein in S. cerevisiae: an unexpected role for copper in iron transportCell76: 393–402, 1994.
27.
Van HoA, WardDM. Kaplan J. Transition metal transport in yeast Annu Rev Microbiol56: 237–261, 2002.
28.
ChungJ, HaileDJ, Wessling-ResnickM. Copper-induced ferroportin-1 expression in J774 macrophages is associated with increased iron efflux. Proc Natl Acad Sci U S A101: 2700–2705, 2004.
29.
ChungJ, ProhaskaJR, Wessling-ResnickM. Ferroportin-1 is not upregulated in copper-deficient mice. J Nutr134: 517–521, 2004
30.
PrasserCL. Adaptalional Biology: Molecules to organismsNew York: John Wiley & Sons, 1986.
31.
GanzoniAM, OakesR, HillmanRS. Red cell aging in vivoJ Clin Invest50:1373–1378, 1971.
32.
GublerCJ, CartwrightGE, WintrobeMM. Studies on copper metabolism, XX: enzyme activities and iron metabolism in copper and iron deficiencies. J Biol Chem224: 533–546, 1957.
33.
CherukuriS. Tripoulas NA, Nurko S, Fox PL. Anemia and impaired stress-induced erythropoiesis in aceruloplasminemic mice Blood Cells Mol Dis33: 346–355. 2004.
34.
Van den BergGJ, Van HouwelingenF, LemmensAG, BeynenAC. Impaired iron status in rats as induced by copper deficiencyBiol Trace Elem Res35:77–79, 1992.
35.
ProhaskaJR. Repletion of copper-deficient mice and brindled mice with copper or iron. J Nutr114: 422–430, 1984.
36.
BingFC, SaurweinEM, MyersVC. Studies in the nutritional anemia of the rat, X: hemoglobin production and iron and copper metabolism with milk of low copper content. J Biol Chem105: 343–3541934
37.
WilliamsDM, KennedyFS, GreenBG. Hepatic iron accumulation in copper-deficient rats. Br J Nutr50: 653–660, 1983.
38.
KarimbakasJ, Langkamp-HenkenB, PercivalSS. Arrested maturation of granulocytes in copper deficient mice. J Nutr128: 1855–18601998
39.
PercivalSS. Neutropenia caused by copper deficiency: possible mechanisms of action. Nutr Rev53: 59–66, 1995.
40.
WilliamsDM, LoukipoulosD. Lee GR, Cartwright GE. Role of copper in mitochondrial iron metabolism. Blood48: 77–851976
41.
WilliamsDM, KennedyFS, GreenBG. The effect of iron substrate on mitochondrial haem synthesis in copper deficiency. Br J Nutr53: 131–136. 1985.
HarrisZL. Durley AP. Man TK, Gitlin JD. Targeted gene disruption reveals an essential role for ceruloplasmin in cellular iron efflux Proc Natl Acad Sci U S A96: 10812–10817. 1999.
